A Compound Heterozygosis of Two Novel Mutations in vWF Exacerbates vWD in a Chinese Pedigree.

BACKGROUND: von Willebrand disease (vWD), caused by mutations in the von Willebrand factor (vWF) coding gene, is a disease characterized by abnormal coagulation activity and a severe tendency for hemorrhage. Therefore, identifying mutations in vWF is important for diagnosing congenital vWD. METHODS: We studied a 23-year-old male vWD patient and his parents. Clotting methods were used to determine activated partial thromboplastin time (aPTT), prothrombin time (PT), fibrinogen (FIB) levels, FVIII activity. Chromogenic substrate method was used to determine vWF antigen and activity. The platelet count was determined. Mutations were searched using whole-exome sequencing and certified by Sanger sequencing. Clinical data, including activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen levels, FX activity, FX antigen levels, and the platelet count were collected. A mixing study was performed to eliminate the presence of coagulation factor inhibitors and lupus anticoagulants. Mutations were screened by using whole-exome sequencing (WES) and were verified by using Sanger sequencing. RESULTS: The proband showed severely decreased vWF antigen, vWF activity, and FVIII activity. RIPA (RISTO-CETIN-induced platelet aggregation) was 0%. Data from WES showed that the proband carried compound heterozygous variants vWF: NM_000552.5 (c.3213C>A p.Cys1071Ter) and vWF: NM_000552.5 (c.6598+2T>C). The proband's mother carried variant vWF: NM_000552.5 (c.3213C>A p.Cys1071Ter) while the proband's father carried variant vWF: NM_000552.5 (c.6598+2T>C). All laboratory test indexes of the proband's parents, including vWF antigen, vWF activity, and FVIII activity, were within the normal ranges. CONCLUSIONS: We identified a compound heterozygosis with two novel mutations in vWF (c.3213C>A, c.6598+2T >C) in a family pedigree, and our results demonstrate that the compound heterozygous mutations probably exacerbate vWD.

Chronic pain accelerates cognitive impairment by reducing hippocampal neurogenesis may via CCL2/CCR2 signaling in APP/PS1 mice.

Patients with chronic pain often have cognitive impairment; this is especially true in elderly patients with neurodegenerative diseases such as Alzheimer's disease (AD), but the mechanism underlying this association remains unclear. This was addressed in the present study by investigating the effect of chronic neuropathic pain on hippocampal neurogenesis and cognitive impairment using amyloid precursor protein/presenilin 1 (APP/PS1) double transgenic mice subjected to spared-nerve injury (SNI). The Von Frey test was performed to determine the mechanical threshold of mouse hind limbs after SNI. The Morris water maze test was used to evaluate spatial learning and memory. Doublecortin-positive (DCX+), 5-bromo-2'-deoxyuridine (BrdU)+, BrdU+/neuronal nuclei (NeuN)+, and C-C motif chemokine ligand 2 (CCL2)+ neurons in the dentate gyrus of the hippocampus were detected by immunohistochemistry and immunofluorescence analysis. CCL2 and C-C chemokine receptor type 2 (CCR2) protein levels in the mouse hippocampus were analyzed by western blotting. The results showed that APP/PS1 mice with chronic neuropathic pain induced by SNI had significant learning and memory impairment. This was accompanied by increased CCL2 and CCR2 expression and decreases in the number of DCX+, BrdU+, and BrdU+/NeuN+ neurons. These results suggest that chronic neuropathic pain is associated with cognitive impairment, which may be caused by CCL2/CCR2 signaling-mediated inhibition of hippocampal neurogenesis. Thus, therapeutic strategies that alleviate neuropathic pain can potentially slow cognitive decline in patients with AD and other neurodegenerative diseases.

Fifteen complete chloroplast genomes of Trapa species (Trapaceae): insight into genome structure, comparative analysis and phylogenetic relationships.

BACKGROUND: Trapa L. is a floating-leaved aquatic plant with important economic and ecological values. However, the species identification and phylogenetic relationship within Trapa are still controversial, which necessitates the need for plastid genome information of Trapa. In this study, complete chloroplast genomes of 13 Trapa species/taxa were sequenced and annotated. Combined with released sequences, comparative analyses of chloroplast genomes were performed on the 15 Trapa species/taxa for the first time. RESULTS: The Trapa chloroplast genomes exhibited typical quadripartite structures with lengths from 155,453 to 155,559 bp. The gene orders and contents within Trapa were conservative, but several changes were found in the microstructure. The intron loss of rpl2, also detected in Lythraceae, was found in all Trapa species/taxa, suggesting close genetic relationship between Lythraceae and Trapaceae. Notably, two small-seed species (T. incisa and T. maximowiczii) showed the smallest genome size with 155,453 and 155,477 bp, respectively. Each cp genome contained the same 130 genes consisting of 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Trapa species/taxa showed 37 (T. incisa and T. maximowiczii) to 41 (T. sibirica) long repeats, including forward, palindromic, reversed and complementary repeats. There were 110 (T. quadrispinosa) to 123 (T. incisa and T. maximowiczii) SSR (simple sequence repeat) loci in Trapa chloroplast genomes. Comparative analyses revealed that two hotspot regions (atpA-atpF and rps2-rpoC2) in Trapa chloroplast genomes could be served as potential molecular markers. Three phylogenetic analyses (ML, MP and BI) consistently showed that there were two clusters within Trapa, including large- and small-seed species/taxa, respectively; for the large-seed Trapa, they clustered according to their geographical origin and tubercle morphology on the surface of seeds. CONCLUSION: In summary, we have acquired the sequences of 13 Trapa chloroplast genomes, and performed the comparative analyses within Trapa for the first time. The results have helped us better identify the Trapa species/taxa and deepen the understanding of genetic basis and phylogenetic relationship of Trapa, which will facilitate the effective management and utilization of the important genetic resources in the future.

Ramsay Hunt syndrome and mandibular alveolar bone necrosis following herpes zoster: A case report and literature review.

Background: Reactivation of latent varicella-zoster virus (VZV) can induce herpes zoster (HZ). Ramsay Hunt syndrome (RHS) occurs through the reactivation and proliferation of VZV in the geniculate ganglion, which can lead to vesicular rash in the ear or oral mucosa, accompanied by neurological disorders. Materials and methods: A 50-year-old man sought a remedy for pain in the right ear and face. Within 1 week, all his lower right teeth fell out, and in the following 3 months, his lower right mandibular alveolar bone gradually became necrotic. In the past 20 days, he experienced blister rash, hearing and taste loss, and slight facial paralysis. Results: After ruling out tumors and other infectious diseases, he was diagnosed with trigeminal HZ and RHS. Conclusion: Ramsay Hunt syndrome with tooth loss and alveolar osteonecrosis is rare. It requires long-term treatment of pain, and prevention and treatment of tooth loss and alveolar bone necrosis are difficult and warrant further study.

The complete chloroplast genome sequence of Trapa incisa Sieb. & Zucc. (Lythraceae).

Trapa L., an annual floating-leaved herb, is widely distributed in the old world and has important edible and medicinal values. However, the taxonomy and phylogeny of Trapa are unclear. Here, we reported the complete chloroplast genome of a wild species with small nuts, T. incisa. The complete chloroplast genome size of T. incisa was 155, 453 bp, consisting of two inverted repeat (IR) regions (24, 388 bp), one large single copy (LSC) region (88, 398 bp) and one small single copy (SSC) region (18, 279 bp). A total of 129 genes were annotated, including 83 protein-coding genes, 38 tRNA genes and 8 rRNA genes. Among them, 19 genes were duplicated (6 protein-coding genes, 9 tRNA genes and 4 rRNA genes). The phylogenomic analysis suggested a close relationship between T. incisa and T. maximowiczii.

The complete chloroplast genome sequence of Trapa kozhevnikoviorum Pshenn. (Lythraceae).

Trapa (Lythraceae) is an economically important aquatic genus used for food and medicine, with wide distribution in Asia, Africa, and Europe. Identification of species, genetic studies and utilization of Trapa are limited by lack of molecular data. Herein, we report the complete chloroplast (cp) genome sequence of a wild species, Trapa kozhevnikoviorum Pshenn. The cp genome size of T. kozhevnikoviorum is 155,545 bp, consisting of a pair of inverted repeat regions (IRa/IRb) of length 24,388 bp, separated by the small single copy (SSC) region of 18,275 bp and a large single copy (LSC) region of 88,494 bp. A total of 113 unique genes, including 79 protein-coding, 30 tRNA, and four rRNA were annotated. Phylogenetic analysis based on 15 whole cp genomes of Lythraceae species supported the monophyletic clustering of Trapa. A cladal relationship among T. kozhevnikoviorum, T. bicornis, and T. natans was revealed.

Infective bicuspid aortic valve endocarditis causing acute severe regurgitation and heart failure: A case report.

BACKGROUND: Infective endocarditis (IE) is an uncommon but potentially life-threatening infection, which occasionally develops into acute severe valve insufficiency leading to the onset of heart failure, and necessitates timely intervention. However, the variable and atypical clinical manifestations always make the early detection of IE difficult and challenging. CASE SUMMARY: A 45-year-old female who was previously healthy presented with exertional shortness of breath and paroxysmal nocturnal dyspnea. She also suffered from a significant decrease in exercise capacity, whereas her body temperature was normal. She had severe hypoxemia and hypotension along with a marked aortic valve murmur. Diffuse pulmonary edema and bilateral pleural effusion were observed on both chest X-ray and computed tomography scan. Transthoracic echocardiography was performed immediately and revealed severe regurgitation of the bicuspid aortic valve. Transesophageal echocardiography was further performed and vegetations were detected. In addition to adequate medical therapy and ventilation support, the patient underwent urgent and successful aortic valve replacement. Her symptoms were significantly relieved and the postoperative chest X-ray showed that pulmonary edema was significantly reduced. Histopathology of the resected valve and positive microorganism culture of the surgical specimen provided evidence of definite IE. CONCLUSION: IE should be considered in critical patients with refractory heart failure caused by severe bicuspid aortic valve regurgitation.

Rivaroxaban plus Percutaneous catheter-directed treatment for a patient with massive pulmonary embolism and colporrhagia.

Massive pulmonary embolism (PE) is defined by acute PE with sustained systemic arterial hypotension that is below 90 mm Hg for at least 15 min or requires inotropic agents (Jaff et al., 2011). For patients with absolute contraindications to thrombolysis, interventional treatment requires the removal of obstructing thrombi from the main pulmonary arteries to facilitate RV recovery and improve symptoms and survival (European Respiratory Society et al., 2014). For patients with acute PE, anticoagulation is recommended, with the objective of preventing both early death and recurrent symptomatic or fatal VTE. Rivaroxaban, an oral factor Xa inhibitor and a new oral anticoagulants, shows effective anticoagulation within hours of administration. It has a fixed-dose regimen, and requires no laboratory monitoring (EINSTEIN-PE Investigators et al., 2012). However, the efficacy and safety of rivaroxaban plus catheter-directed treatment for massive PE and bleeding is unknown. This case demonstrated that a combination of catheter-directed treatment and rivaroxaban was safe and effective in for the treatment of severe PE with vaginal bleeding.

Efficacy and Acceptability of Different Auxiliary Drugs in Pediatric Sevoflurane Anesthesia: A Network Meta-analysis of Mixed Treatment Comparisons.

Emergence agitation preventive medicine should be combined with pediatric anesthesia because of the high frequency of emergence agitation. However, it is challenging to determine the most appropriate medication that can be introduced into pediatric anesthesia for the sake of emergence agitation prevention. We reviewed and retrieved the data from PubMed and Embase. Various medications were assessed based on several endpoints including Emergence agitation outcomes (EA), postoperative nausea and vomiting (PONV), the number of patients who required analgesic (RA), pediatric anesthesia emergence delirium (PAED), the extubation time, the emergency time and the duration of post-anesthesia care unit (PACU) stay. Both traditional and network meta-analysis were carried in this study. A total of 45 articles were complied with the selection criteria and the corresponding articles were reviewed. Fentanyl demonstrated the highest cumulative ranking probability which was followed by those of ketamine and dexmedetomidine with respect to EA and PAED. When PONV and RA were concerned together, clonidine exhibited the highest cumulative ranking probability compared to other medications. Our study suggested that dexmedetomidine perhaps is the most appropriate prophylactic treatment which can be introduced into anesthesia for preventing emergence agitation.

Functional Reorganization of the Primary Somatosensory Cortex of a Phantom Limb Pain Patient.

Functional reorganization of the somatosensory system was widely observed in phantom limb pain patients. Whereas some studies demonstrated that the primary somatosensory cortex (S1) of the amputated limb was engaged with the regions around it, others showed that phantom limb pain was associated with preserved structure and functional organization in the former brain region. However, according to the law of use and disuse, the sensitivity of S1 of the amputated limb to pain-related context should be enhanced due to the adaptation to the long-lasting phantom limb pain experience. Here, we collected neurophysiological data from a patient with 21-year phantom limb pain using electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) techniques. EEG data showed that both laser-evoked potentials (LEPs) and tactile-evoked potentials (TEPs) were clearly presented only when radiant-heat laser pulses and electrical pulses were delivered to the shoulder of the healthy limb, but not of the amputated limb. This observation suggested the functional deficit of somatosensory pathways at the amputated side. FMRI data showed that significant larger brain activations by painful rather than non-painful stimuli in video clips were observed not only at visual-related brain areas and anterior/mid-cingulate cortex, but also at S1 contralateral to the amputated limb. This observation suggested the increased sensitivity of S1 of the amputated limb to the pain-related context. In addition, such increase of sensitivity was significantly larger if the context was associated with the amputated limb of the patient. In summary, our findings provided novel evidence for a possible neuroplasticity of S1 of the amputated limb: in an amputee with long-lasting phantom limb pain, the sensitivity of S1 to pain-related and amputated-limb-related context was greatly enhanced.

[Radiographic anatomical analysis of the pelvic Teepee view].

OBJECTIVES: To research radiographic anatomy of the main structure of the pelvic Teepee view, including its azimuth direction and view anatomy structure. METHODS: From June 2013 to June 2014 adult pelvic CT examination results were filtered, excluding skeletal deformities and pelvic osseous destruction caused by tumors, trauma, etc. The data of 2.0 mm contiguous CT scan of 9 adults' intact pelves was,selected and input into Mimics 10.01 involving 7 males and 2 females with an average age of (41.2±10.3) years old. Utilizing the software, the 3D CT reconstructions of the pelves were completed. Setting the transparency being high,the pelvic 3D reconstructions were manipulated from the pelvic anteroposterior view to the combined obturator oblique outlet view and fine-tuned till the regular Teepee-or teardrop-shaped appearance emerges. Cutting tools of the software were at the moment applied to separate the "Teepee" from the main pelvis for each reconstruction. Then the "Teepee" and the rest (main) part of the pelvis were displayed in different color to facilitate the analysis on the Teepee, iliac-oblique, and anteroposterior views. RESULTS: The "Teepee" started from the posterolateral aspect of the anterior inferior iliac spine and finished at the cortex between the posterior superior iliac spine and the posterior inferior iliac spine in a direction of being from caudal-anterior-lateral to cranial-posterior-medial. The radiographic anatomical composition of the "Teepee" contained one tip, one base,and two aspects. With the inner and outer iliac tables being the inner and outer aspects of the "Teepee", the tip is consequently formed by their intersection. The base is imaged from the cortex of the greater sciatic notch. The medial-inferior-posterior portion of the "Teepee" contains a small part of sacroiliac joint and its corresponding side of bone of the sacrum. CONCLUSIONS: The "Teepee" is a zone of ample osseous structures of the pelvis, aside from a small medial-inferior-posterior portion, the main zone of which can be accepted as a safe osseous zone for the anchor of implants stabilizing certain pelvic and acetabular fracture patterns. The Teepee view can be utilized as guidance for the safe percutaneous insertion of such implants.

Efficacy and safety of remifentanil and sulfentanyl in painless gastroscopic examination: a prospective study.

We aim to assess efficacy and safety of remifentanil or sulfentanyl combined with propofol during painless gastroscopic examination in patients. In this study, 270 patients were randomly divided into 3 groups: propofol was given only in group P; propofol and remifentanil in group PR; propofol and sulfentanyl in group PS during the gastroscopic examination. Efficiency of group P was significantly higher than that of group PR and PS (P<0.01) [corrected]. Efficiency of group PR was lower than that of group PS (P<0.05). Incidence of chest wall rigidity and oxygen desaturation in group PR were higher than group P and PS (P<0.05), whereas there was no difference between groups P and PS (P>0.05). Propofol combined with remifentanil could provide satisfying anesthesia and more respiratory depression, whereas sulfentanyl at equivalent dose combined with propofol could also provide with satisfying anesthesia and less respiratory depression. Combined sufentanyl with propofol would be an effective anesthesia technique in the daytime procedure.

Corticotropin-releasing hormone and urocortin promote phagocytosis of rat macrophages through convergent but distinct pathways.

AIMS: Phagocytosis plays essential roles during inflammation and immune response. This study aims to explore the underlying mechanism of corticotropin-releasing hormone (CRH) and urocortin (UCN)-promoted phagocytosis of rat macrophages. MAIN METHODS: To induce phagocytosis, rat macrophages were incubated with carboxylated fluorescent microspheres. The phagocytosis activity was evaluated by flow cytometric analysis. Actin reorganization was determined by immunostaining with TRITC-labeled phalloidin and transmission electron microscopy (TEM) analysis. Protein expressions of p-RhoA, p-Rac1, p-extracellular signal-related kinase (ERK)1/2 and GAPDH were examined by Western blotting. Protein kinase C (PKC) and protein kinase A (PKA) activities were examined using PreTag non-radio activity assay. KEY FINDINGS: Administration of CRH or UCN alone significantly enhanced phagocytosis of microspheres by rat macrophages, as well as actin reorganization. Ligation of CRH and UCN with CRH receptor increased the phosphorylation of both RhoA and Rac1. Inhibition of RhoA/Rac1 signal pathway suppressed CRH- or UCN-enhanced phagocytosis and actin reorganization. Blockage of PKA signal by MDL-12330A decreased CRH or UCN-promoted p-RhoA and p-Rac1 expressions. Blockage of PKC signal by cholerythine choride decreased CRH or UCN-promoted p-Rac1 expression and UCN-promoted p-RhoA expression, but increased the CRH-induced p-RhoA expression. ERK1/2 was also activated and served as upstream factor of RhoA/Rac1 signal pathway. SIGNIFICANCE: The results reveal that CRH and UCN promote phagocytosis of rat macrophages through convergent but dissociable pathways. PKA/PKC-ERK1/2-RhoA/Rac1 signal pathway plays an essential role in CRH- and UCN-enhanced phagocytosis.

[Influence of the included angle between anterior aspects of S2 and S1 vertebral bodies on pelvic inlet imaging in mid-line sagittal plane].

OBJECTIVE: To analyze the influence of included angle between the anterior aspects of S2 and S vertebral bodies on pelvic inlet imaging in the pelvic midline sagittal plane. METHODS: Totally 58 axial pelvic CT scans were chosen as study objects including 43 males and 15 females,with an average age of 40.7 years old (ranged,18 to 68 years old). The angles between the anterior aspects of S2 and S1, vertebral bodies and the horizontal plane on midline sagittal CT reconstruction were measured to simulate the optimal S2 and S1 inlet angles. The included angle between the anterior aspects of S2 and S1 vertebral bodies was calculated by subtrocting the S1,inlet angle from the S2 inlet angle defined as a base number. Then, the impact of the calculated included angles on the pelvic inlet imaging was analyzed. Results:The S2 inlet angles averaged (30.5±6.5) degrees; the S inlet angles averaged (25.7±5.9) degrees. The difference between them was significant (t=3.35, P=0.001). Ten patients had zero angle between the anterior aspects of S2 and S1 vertebral bodies; 14 patients had negative angle, averaged-(8.9±8.1) degrees; 34 patients had positive angle,averaged (11.8+6.4) degrees. CONCLUSION: The difference of included angle between the anterior aspects of S2 and S1 vertebral bodies leads to the difference between S1 inlet view and S2 inlet view in most cases, complicating the pelvic inlet imaging,and affecting the reliability of the application of pelvic inlet view. Utilizing the angles measured on the preoperative midlihe sagittal CT reconstruction to obatin the patient-customized S1 and S2 inlet views could accurately guide the S1 and S2 iliosacral screw insertion.

[Effect of the anterior aspect of sacral nerve root tunnel on iliosacral screw placement on the standard lateral image of sacrum].

OBJECTIVE: To introduce the location and course of S1, S2 sacral nerve root tunnel and to clarify the significance of the anterior aspect of sacral nerve root tunnel on placement of iliosacral screw on the standard lateral sacral view. METHODS: Firstly the data of 2.0 mm slice pelvic axial CT images were imported into Mimics 10.0, and the sacrum, innominate bones, and sacral nerve root tunnels were reconstructed into 3D views respectively, which were rotated to the standard lateral sacral views, pelvic outlet and inlet views. Then the location and course of the S1, S2 sacral nerve root tunnel on each view were observed. RESULTS: The sacral nerve root tunnel started from the cranial end and anterior aspect of the vertebral canal of the same segment and ended up to the anterior sacral foramen with a direction from cranial-posterior-medial to caudal-anterior-lateral. The tunnel had a lower density than the iliac cortex and greater sciatic notch on the pelvic X-rays,especially on the standard sacral lateral view, on which it showed up as a disrupted are line and required more careful recognition. CONCLUSION: It can prevent the iliosacral screw from penetrating the sacral nerve root tunnel and vertebral canal when recognizing the anterior aspect of sacral nerve root tunnel and choosing it as the caudal-posterior boundary of the "safe zone" on the standard lateral sacral view.

[Classification of upper sacral segment based on continuous axial pelvic computed tomography scan].

OBJECTIVES: To introduce a classification system of upper sacral segment and its significance based on the continuous pelvic axial computed tomography scan. METHODS: The whole pelvis 2.0 mm thick axial scan images of 127 cases were observed, the sacroiliac screw channel of S1 were measured, according to the size of the transverse screw channel the upper sacral segment were classified. Such as transverse screw channel existed and in at least 4 layer scan images its width was > 7.3 mm, it was defined as sacral segment of the normal type. Such as transverse screw channel existed and its maximum width was 7.3 mm or less on scanning level, it was defined as a transitional. Such as transverse channel did not exist, or its width on all scanning level was 0 mm or less, it was defined as dysplastic. Various cases,percentage, and the average of the transverse screw channel were calculated. RESULTS: There were 58 normal (45.7%),42 transitional (33.1%), and 27 dysplastic (21.2%) upper sacral segments with an averaged width of the tansverse screw channel of 13.9 mm, 5.2 mm, and 0.9 mm, respectively. Each specimen could be defined as one of the three types of upper sacral segment without exceptions. CONCLUSION: It is possible to insert a transverse iliosacral screw into a normal upper sacral segment when indicated because of the capacious transverse screw channel. The transverse iliosacral screw placement into the transitional and dysplastic upper sacral segments was contraindicated because of the limited or none transverse screw channel. The transitional upper sacral segment was superior to the dysplastic segment due to its starting point location restriction on the true lateral sacral view.

Lipopolysaccharide upregulates the expression of corticotropin-releasing hormone via MAP kinase pathway in rat peritoneal macrophages.

The stress neuropeptide, corticotropin-releasing hormone (CRH) is expressed in peripheral tissues and inflammatory sites and is implicated in the modulation of the inflammatory response in a paracrine/ autocrine manner. However, the mechanisms by which CRH expression is regulated in peripheral immune cells are unclear. In this article, we address this question by employing primary rat peritoneal macrophages treated with lipopolysaccharide (LPS). Our results showed that CRH could be detected at the mRNA and protein levels in normal peritoneal macrophages and the levels increased significantly and reached a peak at 4 h after stimulation with 100 ng/ml LPS. Furthermore, LPS-induced CRH expression was inhibited by pretreatment with PD98059, a specific MAP kinase inhibitor, in a dose-dependent fashion in which the mRNA and protein levels of CRH was decreased by 90% and 95%, respectively. In addition, pretreatment with 50 µM SB203580, a p38 MAPK inhibitor, led to the decrease of CRH mRNA level by about 41%. Altogether, these results demonstrate that LPS significantly upregulates CRH expression through MAP kinase signaling pathway in rat peritoneal macrophages.

High-dose glucocorticoids induce decreases calcium in hypothalamus neurons via plasma membrane Ca(2+) pumps.

In our previous studies, we occasionally found that high-dose glucocorticoids (GC) induced decrease in [Ca(2+)](i) in hypothalamus neurons. In previous articles, modulation of Ca(2+) channels by GC has been shown to contribute to the elementary regulation of several neuronal functions. However, little is known about the regulation of the Ca efflux pathways that counterbalance the Ca(2+) influx in neurons caused by high-dose GC. In this study, we demonstrate that a high-dose of GC (10 M dexamethasone) caused a 20% decrease in [Ca(2+)](i) within 2 s in cultured hypothalamic neurons; furthermore, we show that an antagonist of the GC receptor blocks this action. To ascertain the temporal sequence of relevant calcium transport mechanisms we selectively blocked the main calcium transporters, including sodium/calcium exchanger (NCX), plasma membrane calcium pumps (PMCA), and P-type Ca(2+)-ATPases of the sarcoplasmic reticulum (SERCA). The GC-induced [Ca(2+)](i) decrease disappeared completely when PMCA was blocked, but not when NCX and SERCA were blocked. These results suggest that high-dose GC (10(-6) M) rapidly decreases [Ca(2+)](i) by activating PMCA but not NCX or SERCA.