A female patient carrying a novel DMD mutation with non-random X-chromosome inactivation from a DMD family.

OBJECTIVE: To analyze the clinical phenotype and genetic characteristics of a female proband carrying a novel mutation in the DMD gene with non-random X-chromosome inactivation in a large pedigree with pseudohypertrophic muscular dystrophy. METHODS: Clinical information of the female proband, her monozygotic twin sister, and other family members were collected. Potential pathogenic variants were detected with Multiplex Ligation-dependent Probe Amplification (MLPA) and whole-exome sequencing (WES). Methylation-sensitive restriction enzyme (HhaI) was employed for X-chromosome inactivation analysis. RESULTS: The proband was a female over 5 years old, displayed clinical manifestations such as elevated creatine kinase (CK) levels and mild calf muscle hypertrophy. Her monozygotic twin sister exhibited normal CK levels and motor ability. Her uncle and cousin had a history of DMD. WES revealed that the proband carried a novel variant in the DMD (OMIM: 300,377) gene: NM_004006.3: c.3051_3053dup; NP_003997.2: p.Tyr1018*. In this pedigree, five out of six female members were carriers of this variant, while the cousin and uncle were hemizygous for this variant. X-chromosome inactivation analysis suggested non-random inactivation in the proband. CONCLUSION: The c.3051_3053dup (p.Tyr1018*) variant in the DMD gene is considered to be the pathogenic variant significantly associated with the clinical phenotype of the proband, her cousin, and her uncle within this family. Integrating genetic testing with clinical phenotype assessment can be a valuable tool for physicians in the diagnosis of progressive muscular dystrophies, such as Becker muscular dystrophy (BMD) and Duchenne muscular dystrophy (DMD).

Reticulocyte hemoglobin content associated with the risk of iron deficiency anemia.

Background/Objective: Reticulocyte hemoglobin content (MCHr) was recognized as a rapid and reliable marker for investigating iron deficiency (ID). We hypothesized that MCHr was associated with the risk of iron deficiency anemia in adults. Methods: This is a dual-center case-control study. A total of 806 patients and healthy individuals were recruited from Ruijin Hospital and Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine between January 2021 and December 2021. The participants were categorized into iron deficiency anemia (IDA) group (n = 302), non-IDA group (n = 366), and healthy control group (n = 138). According to the MCHr level, the participants were divided into two groups, i.e. normal MCHr (≥25 pg) and decreased MCHr (<25 pg) group. Multivariate logistic regression analysis and adjusted subgroup analysis were conducted to estimate the relative risk between MCHr and IDA, with confounding factors including age, sex, hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), Hematocrit (HCT), serum iron (Fe), ferritin (Ferrit), and total iron binding capacity (TIBC). Results: Compared with the non-IDA, the MCHr level with IDA decreased significantly. ROC curve analysis showed that MCHr had the largest area under the AUC curve. After comprehensive adjustment for confounding factors, individuals with normal level of MCHr exhibited a decreased risk of IDA (OR = 0.68 [0.60, 0.77], P < 0.01), while the risk of IDA was up to 5 times higher for those with decreased MCHr. Conclusion: Our findings supported the hypothesis that MCHr was associated with the risk of IDA in adults and could serve as an indicator of IDA severity. MCHr holds clinical value as an auxiliary diagnostic indicator, providing valuable insights into whether invasive examinations are warranted in the assessment of IDA.

Whether Detection of Gene Mutations Could Identify Low- or High-Risk Papillary Thyroid Microcarcinoma? Data from 393 Cases Using the Next-Generation Sequencing.

Objective: The objective of this study is to explore the utilization of next-generation sequencing (NGS) technology in evaluating the likelihood of identifying individuals with papillary thyroid microcarcinoma (PTMC ≤10 mm) who are at high or low risk. Design: NGS was used to analyze 393 formalin-fixed, paraffin-embedded tissues of PTC tumors, all of which were smaller than 15 mm. Results: The study found that bilateralism, multifocality, intrathyroidal spread, and extrathyroidal extension were present in 84 (21.4%), 153 (38.9%), 16 (4.1%), and 54 (13.7%) cases, respectively. Metastasis of cervical lymph nodes was identified in 226 (57.5%) cases and 96 (24.4%) cases with CLNM >5. Out of the total number of cases studied, 8 cases (2.3%) showed signs of tumor recurrence, all of which were localized and regional. Genetic alterations were detected in 342 cases (87.0%), with 336 cases revealing single mutations and 6 cases manifesting compound mutations. 332 cases (84.5%) had BRAFV600E mutation, 2 cases had KRASQ61K mutation, 2 cases had NRASQ61R mutation, 8 cases had RET/PTC1 rearrangement, 3 cases had RET/PTC3 rearrangement, and 1 case had TERT promoter mutation. Additionally, six individuals harbored concurrent mutations in two genes. These mutations were of various types and combinations: BRAFV600E and NRASQ61R (n = 2), BRAFV600E and RET/PTC3 (n = 2), BRAFV600E and RET/PTC1 (n = 1), and BRAFV600E and TERT promoter (n = 1). The subsequent analysis did not uncover a significant distinction in the incidence of gene mutation or fusion between the cN0 and cN1 patient cohorts. The presence of BRAFV600E mutation and CLNM incidence rates were found to be positively correlated with larger tumor size in PTMC. Our data showed that gene mutations did not appear to have much to do with high-risk papillary thyroid microcarcinoma (PTMC). However, when we looked at tumor size, we found that if the tumor was at least 5 millimeters in size, there was a higher chance of it being at high risk for PTM (P < 0.001, odds ratio (OR) = 2.55, 95% confidence interval (CI): 1.57-4.14). Identification of BRAFV600E mutation was not demonstrated to be significantly correlated with advanced clinicopathological characteristics, although it was strongly associated with a bigger tumor diameter (OR = 4.92, 95% CI: 2.40-10.07, P < 0.001). Conclusion: In clinical practice, BRAFV600E mutation does not consistently serve as an effective biomarker to distinguish high-risk PTMC or predict tumor progression. The size of the tumor has a significant correlation with its aggressive characteristics. PTMC with a diameter of ≤5 mm should be distinguished and targeted as a unique subset for specialized treatment.

Fine-Needle Pricking Test of the Parathyroid Gland during Thyroid Surgery in Predicting Parathyroid Function.

Background: Permanent hypoparathyroidism is a serious complication following total thyroidectomy plus central neck dissection (CND). How to evaluate the vascularization of the parathyroid gland in real time is a major concern of thyroid surgeons. This study aimed to evaluate the fine-needle pricking (FNP) test in predicting parathyroid gland function. Methods: The FNP test was performed in patients undergoing total thyroidectomy plus CND between January 1, 2014, and December 31, 2019, to visualize the vascularization of the parathyroid glands. Patients were classified according to the number of parathyroid glands preserved in situ with excellent vascularity (PGPIEV) demonstrated by FNP: group 0 (without PGPIEV), group 1 (with one PGPIEV), group 2 (with two PGPIEV), group 3 (with three PGPIEV), and group 4 (with four PGPIEV). Results: A total of 608 patients with four parathyroid glands underwent FNP testing during thyroidectomy. At least one PGPIEV was demonstrated by FNP testing in 581 patients who had intact parathyroid hormone (iPTH) levels in the normal range after the operation. The prevalence of hypocalcemia decreased from 77.8% in group 0 to 9.8% in group 4 (P < 0.001), and the incidence of hypoparathyroidism decreased from 44.4% in group 0 to 0% in groups 1-4 (P < 0.001). iPTH concentrations on postoperative day 1 were positively correlated with PGPIEV groups (increased from 14.58 ng/l in group 0 to 45.22 ng/l in group 4, P < 0.001). Conclusions: The FNP test is a safe and reliable method to predict parathyroid function. One PGPIEV demonstrated by the FNP test rules out the possibility of patients developing hypoparathyroidism.

The Clinical Significance of Detecting Blood Supply to the Inferior Parathyroid Gland Based on the "Layer of Thymus-Blood Vessel-Inferior Parathyroid Gland" Concept.

Objective: How to preserve the inferior parathyroid gland (IPTG) in situ during central neck dissection (CND) is the major concern of thyroid surgeons. The "layer of thymus-blood vessel-IPTG" (TBP layer) concept showed to be effective in preserving IPTG. The objective of this study was to identify the origin and course of blood supply to IPTG (IPBS) within the TBP layer and to take key points of operation during CND. Design: This is a retrospective control study. Participants. Patients who underwent thyroidectomy plus CND using the TBP layer concept and conventional technique between 2017 and 2019 were enrolled. Measurements. The origin and course of IPBS in relation to recurrent laryngeal nerve (RLN) and thymus and prevalence of hypoparathyroidism were detected. Results: A total of 71.3% of IPTGs (251 of 352) were supplied by ITA branches, defined as type A. Type A was further divided into Types A1 (branches of ITA, coursing laterally to the RLN (53.1%, 187 of 352)) and A2 (branches of ITA, traversing medially to the RLN (18.2%, 64 of 352)). Type A2 was more common on the right side than on the left side (P < 0.001). Fifty-five (15.6%) IPTG feeding vessels originated from the thymus or mediastinum. Nineteen (5.4%) IPTGs were supplied by branches of the superior thyroid artery. The incidence of transient hypoparathyroidism decreased from 45.7% to 3.6% (P < 0.001), in the TBP layer group compared with the conventional technique group. Conclusion: The origin and course of IPBS follow a definite pattern. This mapping and precautions help surgeons optimize intraoperative manipulations for better preservation of IPBS during CND.

Predicting non-small-volume central lymph node metastases (>5 or ≥2 mm) preoperatively in cN0 papillary thyroid microcarcinoma without extrathyroidal extension.

The ability to identify patients with aggressive papillary thyroid microcarcinoma (PTMC) from the low-risk patients is critical to planning proper management of PTMC. Lymph node metastases showed association with recurrence and low survival rate, especially in patients with >5 or ≥2 mm metastatic lymph nodes. Therefore, this study aimed to investigate the preoperatively predictive factors of non-small-volume (metastatic lymph nodes >5 or ≥2 mm in size) central lymph node metastases (NSVCLNM) in PTMC patients. A total of 420 patients with clinically node-negative (cN0) PTMC without extrathyroidal extension underwent thyroidectomy plus central neck dissection (CND) between January 2013 and December 2015, were retrospectively analyzed. Of the 420 patients, 33 (7.9%) had NSVCLNM. The 5-year recurrence-free survival was significantly less in cN0 PTMC patients with NSVCLNM, when compared with patients without NSVCLNM (80.8% vs 100%, P < .001). Multivariate logistic regression revealed age ≤36 years (P < .001), male sex (P = .002), ultrasonic tumor sizes of >0.65 cm (P < .001), and ultrasonic multifocality (P = .039) were independent predictive factors of NSVCLNM. A prediction equation (Y = 1.714 × age + 1.361 × sex + 1.639 × tumor size + 0.842 × multifocality -5.196) was developed, with a sensitivity (69.7%) and a specificity (84.0%), respectively, at an optimal cutoff point of -2.418. In conclusion, if the predictive value was >-2.418 according to the equation, immediate surgery including CND rather than active surveillance might be considered for cN0 PTMC patients.

Prediction mode of more than 5 central lymph nodes metastases in clinically node-negative ipsilateral papillary thyroid carcinoma with tumor size 1 to 4 cm.

According to the 2015 American Thyroid Association guidelines, either lobectomy or total thyroidectomy was recommended for patients with papillary thyroid carcinoma (PTC) of 1 to 4 cm without extrathyroidal extension and lymph node metastasis. However, lymph node metastases showed strong association with recurrence and low survival rate, especially in PTC patients with more than 5 metastatic lymph nodes. Therefore, this study aimed to investigate the predictive factors of more than 5 central lymph nodes metastases (CLNM) in PTC patients with tumor sizes of 1 to 4 cm. A total of 382 patients with clinically node-negative (cN0) ipsilateral PTC who underwent thyroidectomy with central neck dissection between January 2012 and December 2016 were retrospectively analyzed. CLNMs of >5 were found in 54 (14.1%) patients, while CLNM was detected in 230 (60.2%) patients. Multivariate logistic regression revealed age < 45 years (P < .001), male gender (P = .013), and tumor sizes of >2 cm (P = .001) as independent predictive factors of >5 CLNMs in cN0 ipsilateral PTC patients with tumor sizes 1 to 4 cm. The prediction equation (Y = 1.694 × age + 0.807 × gender + 1.190 × tumor size - 3.530) was developed, with a sensitivity (57.4%) and a specificity (80.8%), respectively, at an optimal cut-off point of -1.685. Therefore, if the predictive value was higher than -1.685 according to the equation in cN0 ipsilateral PTC patients with tumor sizes 1 to 4 cm, then total thyroidectomy might be considered.

Anatomical localization of normal parathyroid glands before thyroidectomy through ultrasonography reduces postoperative hypoparathyroidism.

Postoperative hypoparathyroidism due to dysfunction of the parathyroid gland is the most common complication after thyroidectomy. Our objective was to introduce the method of anatomical localization of normal parathyroid glands before thyroidectomy through ultrasonography and to evaluate its efficiency. The study group included 52 patients subjected to anatomical localization of the parathyroid gland prethyroidectomy through ultrasonography. The control group included 52 sex- and age-matched patients without parathyroid gland localization. The proportion of parathyroid glands preserved in situ and postoperative hypoparathyroidism rates in the 2 groups were compared. The rates of normal parathyroid glands identified according to ultrasonography for left superior, left inferior, right superior, and right inferior glands were 78.8%, 90.4%, 57.7%, and 82.7%, respectively. The rate of parathyroid gland excised inadvertently was significantly decreased (P = .038) in the study group as compared with the control group. The rates of parathyroid gland preservation in situ were significantly improved in the left superior (P = .001), left inferior (P = .002), and right inferior glands (P = .005). Furthermore, the incidence of transient hypoparathyroidism decreased significantly (P = .028). Our study indicated that normal parathyroid glands were identified by ultrasonography, and the anatomical localization improved the rate of parathyroid gland preservation in situ and decreased the incidence of transient hypoparathyroidism.

Predictive factors for non-small-volume central lymph node metastases (more than 5 or ≥ 2 mm) in clinically node-negative papillary thyroid carcinoma.

The benefits of prophylactic central neck dissection (pCND) for treating patients with clinical node-negative (cN0) papillary thyroid carcinoma (PTC) remain controversial. Lymph node metastases have been strongly associated with local recurrence and low survival, especially in PTC patients with 5 or more or ≥2 mm metastatic lymph nodes. The following study investigates the incidence and risk factors of more than 5 or ≥2 mm metastatic lymph nodes in the central compartment.A total of 611 patients with cN0 PTC were retrospectively analyzed. Cervical lymph nodes were harvested, and the size of metastatic lymph nodes was consequently analyzed.Non-small-volume central lymph node metastases (NSVCLNM), defined as more than 5 or ≥2 mm metastatic lymph nodes) were detected in 67 (11.0%) patients. Male gender, age ≤36 years, multifocal lesions, extrathyroidal extension, and tumor size > 0.85 cm were independent risk factors for NSVCLNM in cN0 PTC. The sensitivity and specificity of having ≥3 risk factors for predicting NSVCLNM was 46.3% and 86.8%, respectively.These findings suggest that pCND is a suitable treatment strategy for cN0 PTC patients with 3 or more risk factors for NSVCLNM.

Association between decreased serum parathyroid hormone after total thyroidectomy and persistent hypoparathyroidism.

BACKGROUND: Postoperative hypocalcemia caused by hypoparathyroidism is one of the most common morbidities of total thyroidectomy. The aim of this study was to analyze the kinetics and factors affecting PTH levels after total thyroidectomy and central neck dissection (CND). MATERIAL/METHODS: We performed a retrospective study in 438 consecutive patients who underwent total thyroidectomy between January 2007 and December 2010. No patient had a history of thyroid or neck surgery. PTH and calcium levels were recorded 1 day before the operation, during the first 5 days, and during follow-up (2 weeks and 2, 6, and 12 months). RESULTS: PTH levels declined to 41.90% of its initial value on the first day after the operation. After surgery, PTH was correlated positively with calcium and inversely with phosphate levels from postoperative day 1 to 14. Based on clinical observation, using a PTH threshold of <7 ng/L on postoperative day 1 was predictive of persistent hypoparathyroidism, with sensitivity and negative predictive value 100%, but poor specificity (70.19%). CND increased the risk of transient hypoparathyroidism compared with total thyroidectomy alone. Patients with thyroiditis had an increased risk of permanent hypoparathyroidism compared with those without thyroiditis. Iatrogenic removal of the parathyroid glands increased the risk of permanent hypoparathyroidism compared with those without iatrogenic parathyroidectomy. CONCLUSIONS: PTH declined on the first day after thyroidectomy. PTH levels <7 ng/L on the first day after surgery might be associated with persistent hypoparathyroidism. CND, thyroiditis, and iatrogenic parathyroidectomy increased the risk of hypoparathyroidism.

[Research progress--the role of astrocyte in neuronal functions].

Astrocytes can regulate synaptic transmission by releasing gliotransmitter, and also can promote synaptogenesis and neurogenesis by releasing estrogen, thrombospondins, IL-1beta and IL-6. Astrocytes may play critical roles in neural nutrition and neuroprotection, so that it might be a new target for treatment of certain central nervous system diseases.