To analyze the incidence and patterns of calcification of papillary thyroid microcarcinoma (PTMC) on neck ultrasonography (NUS) and assess the clinical implications of calcification, especially for neck node metastasis.
The clinical data of 379 patients with PTMC who underwent thyroidectomy between January and December 2011 were retrospectively analyzed. PTMC lesions were classified into four subgroups according to their calcification patterns on preoperative NUS: microcalcification, macrocalcification, rim calcification, and noncalcification. The clinicopathologic characteristics were compared between the patients with and without calcification, and among the four subgroups.
Calcifications were detected on NUS in 203 patients (53.5%) and central neck node metastasis was observed in 119 patients (31.3%). Calcification was associated with larger tumor size (0.68 cm vs. 0.54 cm), higher rate of lymph node metastasis (38.6% vs. 23.2%) and higher lymph node ratio (0.11 vs. 0.06) compared to noncalcification (All P < 0.05). In addition, the extent of calcification correlated with lesion size (0.67 cm vs. 0.69 cm vs. 0.85 cm). Further, the likelihood of lymph node metastasis also correlated with the extent of calcification in the order of non-, micro- and macrocalcification (23.3%, 36.8%, and 44.1%, respectively). The calcification rate was higher in patients with lymph node metastasis than those without it (65.5% vs.47.7%) (All P < 0.05).
PTMC patients positive for calcification on NUS had a higher rate of lymph node metastasis, and a higher lymph node ratio compared to noncalcification patients. Calcification patterns should be assessed carefully in patients with PTMC by preoperative NUS.
Papillary thyroid microcarcinoma; Calcification; Ultrasonography
The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.
brain; Cre; CaMKII alpha; Gnao; testis
Reelin, an extracellular glycoprotein has an important role in the proper migration and positioning of neurons during brain development. Lack of reelin causes not only disorganized lamination of the cerebral and cerebellar cortex but also malpositioning of mesencephalic dopaminergic (mDA) neurons. However, the accurate role of reelin in the migration and positioning of mDA neurons is not fully elucidated. In this study, reelin-deficient reeler mice exhibited a significant loss of mDA neurons in the substantia nigra pars compacta (SNc) and a severe alteration of cell distribution in the retrorubal field (RRF). This abnormality was also found in Dab1-deficinet, yotari mice. Stereological analysis revealed that total number of mDA neurons was not changed compared to wild type, suggesting that the loss of mDA neurons in reeler may not be due to the neurogenesis of mDA neurons. We also found that formation of PSA-NCAM-positive tangential nerve fibers rather than radial glial fibers was greatly reduced in the early developmental stage (E14.5) of reeler. These findings provide direct evidence that the alteration in distribution pattern of mDA neurons in the reeler mesencephalon mainly results from the defect of the lateral migration using tangential fibers as a scaffold.
Reelin; dopaminergic neurons; radial glia; tangential fibers; neuronal migration
Due to the increased prevalence of thyroid cancer, it has been frequently detected in breast cancer patients recently. The aim of this study was to evaluate the clinicopathologic characteristics of thyroid cancer in breast cancer patients with respect to prognosis and treatment.
From August 1998 to September 2012, 101 breast cancer patients were diagnosed with thyroid cancer (BT group). One hundred ninety-three female patients with a thyroid malignancy that underwent thyroidectomy in 2008 were recruited as controls (oT group). The clinicopathologic results of these two groups were compared.
Patients were older (51.40 vs. 47.16, P < 0.001), mean tumor size was smaller (0.96 cm vs. 1.43 cm, P < 0.001), and extrathyroidal extension was less common in the BT group. In both groups, papillary thyroid carcinoma was the most common type of thyroid malignancy. T and N classifications of thyroid cancer were less severe in the BT group, but group TNM stages were similar. Endoscopic thyroid surgery was performed in 12.9% of patients in the BT group and in 6.7% of patients in the oT group. Postoperative radioactive iodine ablation was performed less often in the BT group (P < 0.001). Group recurrence rates were not significantly different.
Thyroid cancer in breast cancer patients was diagnosed at earlier status than ordinary thyroid cancer. However, the prognosis of thyroid cancer in breast cancer patients was not superior to that in patients with thyroid cancer alone. Radioactive iodine ablation was performed less often and endoscopic surgery could be performed in breast cancer patients.
Breast neoplasms; Prognosis; Second primary neoplasms; Thyroid neoplasms
This study was performed to investigate the effect of desalinated underground seawater (named as 'magma seawater', MSW) of Jeju Island in Korea on lipid metabolism and antioxidant activity. MSW was collected from underground of Han-Dong in Jeju Island, and freely given to high fat diet (HFD)-fed C57BL/6 mice for 10 weeks. Although there were no significant differences in the body weight changes and plasma lipid levels, hepatic triglyceride levels were significantly lower in the MSW group than in the normal tap water (TW)-drunken control group. Furthermore, the activity of fatty acid synthase (FAS) was significantly decreased and carnitine palmitoyltransferase (CPT) activity was increased in MSW group compared to TW group. Similarly, real-time PCR analysis revealed that mRNA expressions of lipogenic genes were lowered in MSW groups compared to the control group. In a morphometric observation on the liver tissue, accumulation of fats was remarkably reduced in MSW group. Meanwhile, in vitro assay, free radical scavenging activity measured by using diphenylpicrylhydrazyl (DPPH) was increased in MSW group. The 2'-7'-dichlorofluorescein diacetate (DCF-DA) staining followed with fluorescent microscopy showed a low intensity of fluorescence in MSW-treated HepG2 cells, compared to TW-treated HepG2 cells, which indicated that the production of reactive oxygen species by tert-butyl hydroperoxide (t-BHP) in HepG2 cells was decreased by MSW treatment. The antioxidant effect of MSW on t-BHP-induced oxidative stress in HepG2 cells was supported by the increased activities of intracellular antioxidant enzymes such as catalase and glutathione reductase. From these results, we speculate that MSW has an inhibitory effect on lipogenesis in liver and might play a protective role against cell damage by t-BHP-induced oxidative stress.
Underground seawater; lipid metabolism; hepatic lipid; antioxidant effect
Stroke is one of the common causes of death and disability. Despite extensive efforts in stroke research, therapeutic options for improving the functional recovery remain limited in clinical practice. Experimental stroke models using genetically modified mice could aid in unraveling the complex pathophysiology triggered by ischemic brain injury. Here, we optimized the procedure for generating mouse stroke model using an intraluminal suture in the middle cerebral artery and verified the blockage of blood flow using indocyanine green coupled with near infra-red radiation. The first week after the ischemic injury was critical for survivability. The survival rate of 11% in mice without any treatment but increased to 60% on administering prophylactic antibiotics. During this period, mice showed severe functional impairment but recovered spontaneously starting from the second week onward. Among the various behavioral tests, the pole tests and neurological severity score tests remained reliable up to 4 weeks after ischemia, whereas the rotarod and corner tests became less sensitive for assessing the severity of ischemic injury with time. Further, loss of body weight was also observed for up 4 weeks after ischemia induction. In conclusion, we have developed an improved approach which allows us to investigate the role of the cell death-related genes in the disease progression using genetically modified mice and to evaluate the modes of action of candidate drugs.
blood flow; middle cerebral artery; survival; stroke; brain ischemia; behavior
Human mesenchymal stem cells (MSCs) have emerged as attractive cellular vehicles
to deliver therapeutic genes for ex-vivo therapy of diverse diseases;
this is, in part, because they have the capability to migrate into tumor or
lesion sites. Previously, we showed that MSCs could be utilized to deliver a
bacterial cytosine deaminase (CD) suicide gene to brain tumors. Here we
assessed whether transduction with a retroviral vector encoding CD gene
altered the stem cell property of MSCs. MSCs were transduced at passage 1 and
cultivated up to passage 11. We found that proliferation and differentiation
potentials, chromosomal stability and surface antigenicity of MSCs were not
altered by retroviral transduction. The results indicate that retroviral vectors
can be safely utilized for delivery of suicide genes to MSCs for
ex-vivo therapy. We also found that a single retroviral
transduction was sufficient for sustainable expression up to passage 10. The
persistent expression of the transduced gene indicates that transduced MSCs
provide a tractable and manageable approach for potential use in allogeneic
ex-vivo therapy; gene therapy; mesenchymal stem cell; retrovirus; safety; suicide gene
During shoulder surgery, blood pressure is frequently measured at the ankle. Anesthetic complications may result when ankle blood pressure is higher than brachial blood pressure and anesthesiologists misinterpret ankle blood pressure as brachial blood pressure. Therefore, we investigated whether ankle blood pressure is significantly higher than brachial blood pressure before anesthesia induction, during induction, after tracheal intubation, before beach chair position, and in the beach chair position.
Thirty patients requiring general anesthesia for shoulder surgery were included in this study. Ankle and brachial blood pressure were simultaneously measured before induction, during induction, after intubation, before beach chair position, and in the beach chair position.
Ankle blood pressure was higher than brachial blood pressure before induction, during induction, after intubation, before beach chair position, and in the beach chair position. Ankle-brachial blood pressure differences in the beach chair condition were much higher than in four other conditions. The correlation coefficient between mean ankle-brachial blood pressure differences before the beach chair position and mean ankle-brachial blood pressure differences in the beach chair position was 0.616. Brachial systolic blood pressure could be predicted by regression equations (R2 = 0.306-0.771).
These results suggest that anesthesiologists should consider these ankle-brachial blood pressure differences when monitoring anesthesia in the beach chair position.
Ankle blood pressure; Ankle-brachial blood pressure index; Beach chair position; Blood pressure; Brachial blood pressure; Shoulder surgery
We report two cases of follicular adenoma of the thyroid with extensive extracellular mucin deposition. Fine needle aspiration in Case 1 showed singly discohesive polygonal cells in a granular mucinous background. They contained abundant eosinophilic cytoplasm, nuclear irregularities, and frequent nuclear inclusions with occasional bizarre mitoses. A right lobectomy was done. In Case 2, a 47-year-old Caucasian woman with multinodular goiter had total thyroidectomy and a yellow-tan nodule was found within the right lobe. Both tumors were well-encapsulated masses with thick capsules. Each was characterized by microfollicles without papillae in a mucinous stroma. Tumor cells were positive for thyroglobulin and negative for calcitonin, CEA, galectin-3, HBME-1, and CK19. The extracellular mucin stained with Alcian-blue and colloidal iron but not with mucicarmine and D-PAS. No BRAF gene mutation was detected. Because there were neither capsular nor vascular invasions, both cases were diagnosed as follicular adenomas of the thyroid with extensive extracellular mucin deposition, which as proposed by the WHO classification can be categorized as a mucinous variant of follicular adenoma. Retrospectively, frequent nuclear inclusions and the absence of nuclear grooves in the mucin-containing background of cytologic smears and histologic sections were shared by those of mucin-producing papillary carcinoma. It is unclear whether it belongs to an existing category of thyroid neoplasm with mucin production or whether it is truly a new tumor variant. Furthermore, pathologists should pay attention to avoid misdiagnosis of this variant of follicular neoplasm that shows an overlapping cytology with that of papillary carcinoma.
thyroid; mucin; follicular adenoma; aspiration; fine needle
CREB mediates the transcriptional effects of glucose and incretin hormones in insulin-target cells and insulin-producing β-cells. Although the inhibition of CREB activity is known to decrease the β-cell mass, it is still unknown what factors inversely alter the CREB signaling pathway in β-cells. Here, we show that β-cell dysfunctions occurring in chronic hyperglycemia are not caused by simple inhibition of CREB activity but rather by the persistent activation of CREB due to decreases in protein phophatase PP2A. When freshly isolated rat pancreatic islets were chronically exposed to 25 mM (high) glucose, the PP2A activity was reduced with a concomitant increase in active pCREB. Brief challenges with 15 mM glucose or 30 µM forskolin after 2 hour fasting further increased the level of pCREB and consequently induced the persistent expression of ICER. The excessively produced ICER was sufficient to repress the transcription of NeuroD, insulin, and SUR1 genes. In contrast, when islets were grown in 5 mM (low) glucose, CREB was transiently activated in response to glucose or forskolin stimuli. Thus, ICER expression was transient and insufficient to repress those target genes. Importantly, overexpression of PP2A reversed the adverse effects of chronic hyperglycemia and successfully restored the transient activation of CREB and ICER. Conversely, depletion of PP2A with siRNA was sufficient to disrupt the negative feedback regulation of CREB and induce hyperglycemic phenotypes even under low glucose conditions. Our findings suggest that the failure of the negative feedback regulation of CREB is the primary cause for β-cell dysfunctions under conditions of pathogenic hyperglycemia, and PP2A can be a novel target for future therapies aiming to protect β-cells mass in the late transitional phase of non-insulin dependent type 2 diabetes (NIDDM).
Most of thyroid lymphomas are B-lineage, and T-cell lymphomas are rare. Here, we report a case of primary thyroid T-cell lymphoma associated with Hashimoto's thyroiditis. A 48-yr-old woman presented with incidentally found neck mass. Histologically, the resected right lobe of the thyroid was replaced by monomorphic small atypical lymphoid cells with lymphoepithelial lesion-like change, most of which were immunoreactive for CD3, CD8, βF-1, and TIA-1. Peripheral T-cell lymphoma, unspecified, was finally diagnosed after molecular study for TCR-γ gene rearrangement. This is the second case of cytotoxic T-cell lymphoma reported in the thyroid gland so far. Unique association between thyroid follicles and neoplastic lymphocytes may be characteristic feature of this type of T-cell lymphoma.
Lymphoma, T-Cell; Thyroid Gland; T-Lymphocytes, Cytotoxic; Lymphoma, B-Cell, Marginal Zone
Disabled 1 (Dab1), a cytoplasmic adaptor protein expressed predominantly in the CNS, transduces a Reelin-initiated signaling that controls neuronal migration and positioning during brain development. To determine the role of Dab1 in neural stem cell (NSC) differentiation, we established a culture of neurospheres derived from the embryonic forebrain of the Dab1−/− mice, yotari. Differentiating Dab1−/− neurospheres exhibited a higher expression of GFAP, an astrocytic marker, at the expense of neuronal markers. Under Dab1-deficient condition, the expression of NeuroD, a transcription factor for neuronal differentiation, was decreased and the JAK-STAT pathway was evidently increased during differentiation of NSC, suggesting the possible involvement of Dab1 in astrocyte differentiation via JAK-STAT pathway. Notably, expression of neural and glial markers and the level of JAK-STAT signaling molecules were not changed in differentiating NSC by Reelin treatment, indicating that differentiation of NSC is Reelin-independent. Immunohistochemical analyses showed a decrease in the number of neurons and an increase in the number of GFAP-positive cells in developing yotari brains. Our results suggest that Dab1 participates in the differentiation of NSCs into a specific cell lineage, thereby maintaining a balance between neurogenesis and gliogenesis.
Disabled 1 Reelin; Astrocyte; Neuron; NeuroD; JAK-STAT; Neural stem cells
Mesenchymal stem cells (MSCs) secrete bioactive factors that exert diverse responses in vivo. In the present study, we explored mechanism how MSCs may lead to higher functional recovery in the animal stroke model. Bone marrow-derived MSCs were transplanted into the brain parenchyma 3 days after induction of stroke by occluding middle cerebral artery for 2 h. Stoke induced proliferation of resident neural stem cells in subventricular zone. However, most of new born cells underwent cell death and had a limited impact on functional recovery after stroke. Transplantation of MSCs enhanced proliferation of endogenous neural stem cells while suppressing the cell death of newly generated cells. Thereby, newborn cells migrated toward ischemic territory and differentiated in ischemic boundaries into doublecortin+ neuroblasts at higher rates in animals with MSCs compared to control group. The present study indicates that therapeutic effects of MSCs are at least partly ascribed to dual functions of MSCs by enhancing endogenous neurogenesis and protecting newborn cells from deleterious environment. The results reinforce the prospects of clinical application using MSCs in the treatment of neurological disorders.
bromodeoxyuridine; doublecortin protein; mesenchymal stem cells; mesenchymal stem cell transplantation; stroke
The profile of membrane currents was investigated in differentiated neuronal cells derived from human neural stem cells (hNSCs) that were obtained from aborted fetal cortex. Whole-cell voltage clamp recording revealed at least 4 different currents: a tetrodotoxin (TTX)-sensitive Na+ current, a hyperpolarization-activated inward current, and A-type and delayed rectifier-type K+ outward currents. Both types of K+ outward currents were blocked by either 5 mM tetraethylammonium (TEA) or 5 mM 4-aminopyridine (4-AP). The hyperpolarization-activated current resembled the classical K+ inward current in that it exhibited a voltage-dependent block in the presence of external Ba2+ (30µM) or Cs+ (3µM). However, the reversal potentials did not match well with the predicted K+ equilibrium potentials, suggesting that it was not a classical K+ inward rectifier current. The other Na+ inward current resembled the classical Na+ current observed in pharmacological studies. The expression of these channels may contribute to generation and repolarization of action potential and might be regarded as functional markers for hNSCs-derived neurons.
Human neural stem cells; TTX-sensitive Na+ current; A-type; delayed rectifier; hyperpolarization-activated inward current
Patients with metastatic breast cancer (MBC) are frequently exposed to high cumulative doses of anthracyclines and are at risk of resistance and cardiotoxicity. This phase II trial evaluated the efficacy and toxicity of docetaxel plus cisplatin, as salvage chemotherapy in patients with MBC resistant to prior anthracyclines.
Patients with MBC that had progressed after at least one prior chemotherapy regimen containing anthracyclines received docetaxel 75 mg/m2 followed by cisplatin 60 mg/m2 every 3 weeks for a maximum of 6 cycles or until disease progression.
Between Jan 2000 and May 2002, 24 patients with tumors primary resistant and 15 with secondary resistant disease were accrued. All 39 patients were evaluable for safety and 36 for efficacy. The objective response rate was 31% (95% CI, 16–45%) with 3 complete responses. The median time to disease progression was 7 months, and the median overall survival was 23 months (median follow-up of 41 months). Neutropenia was the most frequently observed severe hematologic toxicity (39% of patients), whereas asthenia and nausea were the most common non-hematologic toxicities. No treatment-related death was observed.
In conclusion, we found docetaxel plus cisplatin to be an active and safe chemotherapy regimen for patients with MBC resistant to anthracyclines.