Association between calcitonin receptor gene polymorphisms and calcium stone urolithiasis: A meta-analysis

ABSTRACT Purpose It has been reported that calcitonin receptor (CALCR) gene polymorphisms might be associated with calcium stone urolithiasis. Owing to mixed and inconclusive results, we conducted a meta-analysis to summarize and clarify this association. Materials and Methods A systematic search of studies on the association between CALCR gene polymorphisms and calcium stone urolithiasis susceptibility was conducted in databases. Results Odds ratios and 95% confidence intervals were used to pool the effect size. Five articles were included in our meta-analysis. Conclusions CALCR rs1801197 might be associated with increased risk of calcium stone urolithiasis. There is insufficient data to fully confirm the association between CALCR rs1042138 and calcium stone urolithiasis susceptibility. Well-designed studies with larger sample size and more subgroups are required to validate the risk identified in the current meta-analysis.

Effect of remifentanil on pre-osteoclast cell differentiation in vitro

BACKGROUND: The structure and function of bone tissue is maintained through a constant remodeling process, which is maintained by the balance between osteoblasts and osteoclasts. The failure of bone remodeling can lead to pathological conditions of bone structure and function. Remifentanil is currently used as a narcotic analgesic agent in general anesthesia and sedation. However, the effect of remifentanil on osteoclasts has not been studied. Therefore, we investigated the effect of remifentanil on pre-osteoclast (pre-OCs) differentiation and the mechanism of osteoclast differentiation in the absence of specific stimulus. METHODS: Pre-OCs were obtained by culturing bone marrow-derived macrophages (BMMs) in osteoclastogenic medium for 2 days and then treated with various concentration of remifentanil. The mRNA expression of NFATc1 and c-fos was examined by using real-time PCR. We also examined the effect of remifentanil on the osteoclast-specific genes TRAP, cathepsin K, calcitonin receptor, and DC-STAMP. Finally, we examined the influence of remifentanil on the migration of pre-OCs by using the Boyden chamber assay. RESULTS: Remifentanil increased pre-OC differentiation and osteoclast size, but did not affect the mRNA expression of NFATc1 and c-fos or significantly affect the expression of TRAP, cathepsin K, calcitonin receptor, and DC-STAMP. However, remifentanil increased the migration of pre-OCs. CONCLUSIONS: This study suggested that remifentanil promotes the differentiation of pre-OCs and induces maturation, such as increasing osteoclast size. In addition, the increase in osteoclast size was mediated by the enhancement of pre-OC migration and cell fusion.

Study on mechanism of Drynariae Rhizoma in treating osteoporosis with integrative pharmacology perspective / 中国中药杂志

Drynariae Rhizoma has great significance in the clinical practice of osteoporosis treatment. Based on the perspective of integrative pharmacology, the study explored the mechanism of action of Drynariae Rhizoma in the treatment of osteoporosis. Six active components in Drynariae Rhizoma were obtained, mainly including glycosides and sterols. Taking the median of 2 times of "node connectivity" as the card value, the core node of the Chinese medicine target disease gene interaction network was selected. Based on this, three topological structural eigenvalues, such as "node connectivity" "node tightness" and "node connectivity" were calculated, thereby screening out four core targets of Drynariae Rhizoma treatment for osteoporosis, including thyroid parathyroid hormone 1 receptor (PTH1R), parathyroid hormone 2 receptor (PTH2R), calcitonin receptor gene (CALCR), and SPTBN1 gene (SPTBN1). Based on the gene ontology database GO and KEGG pathway database, the molecular function, intracellular localization, and biological reactions and pathways of proteins encoded by drug target genes were determined. Combined with enrichment calculation, it is predicted that osteoporosis may play a role in biosynthetic processes, such as circulatory system, nervous system, energy metabolism, prolactin signal pathway, GnRH signaling pathway, neurotrophic factor signaling pathway and other pathway. The conclusion of this study is certain with the existing research results, and the new target and new pathway could also be used as a theoretical basis for the further verification of osteoporosis.

Compressive force regulates ephrinB2 and EphB4 in osteoblasts and osteoclasts contributing to alveolar bone resorption during experimental tooth movement / 대한치과교정학회지

OBJECTIVE: To investigate the involvement of ephrinB2 in periodontal tissue remodeling in compression areas during orthodontic tooth movement and the effects of compressive force on EphB4 and ephrinB2 expression in osteoblasts and osteoclasts. METHODS: A rat model of experimental tooth movement was established to examine the histological changes and the localization of ephrinB2 in compressed periodontal tissues during experimental tooth movement. RAW264.7 cells and ST2 cells, used as precursor cells of osteoclasts and osteoblasts, respectively, were subjected to compressive force in vitro. The gene expression of EphB4 and ephrinB2, as well as bone-associated factors including Runx2, Sp7, NFATc1, and calcitonin receptor, were examined by quantitative real-time polymerase chain reaction (PCR). RESULTS: Histological examination of the compression areas of alveolar bone from experimental rats showed that osteoclastogenic activities were promoted while osteogenic activities were inhibited. Immunohistochemistry revealed that ephrinB2 was strongly expressed in osteoclasts in these areas. Quantitative real-time PCR showed that mRNA levels of NFATc1, calcitonin receptor, and ephrinB2 were increased significantly in compressed RAW264.7 cells, and the expression of ephrinB2, EphB4, Sp7, and Runx2 was decreased significantly in compressed ST2 cells. CONCLUSIONS: Our results indicate that compressive force can regulate EphB4 and ephrinB2 expression in osteoblasts and osteoclasts, which might contribute to alveolar bone resorption in compression areas during orthodontic tooth movement.

Correlación de los niveles séricos de procalcitonina con el curso clínico de la sepsis / Correlation of procalcitonin serum levels with the clinical course of sepsis

Objetivo: observar el comportamiento de los niveles séricos de procalcitonina durante el curso clínico de la sepsis en la población local. Metodología: estudio de cohorte prospectivo. Se ingresaron al estudio pacientes admitidos a la unidad de cuidados intensivos del hospital clínica Kennedy, sede Policentro, con diagnóstico de sepsis y sin tratamiento antibiótico por mas de 48h previas al ingreso. Se obtuvo valores séricos de procalcitonina (PCT), proteína C reactiva (PCR) y se calcularon los scores APACHE II y MODS al momento del ingreso y diariamente hasta la resolución del cuadro séptico o muerte del paciente. Resultados: ocho pacientesfueron incluidos en el estudio. Se observó una mayor disminución porcentual de PCT en sobrevivientes que no-sobrevivientesentre D1-D2 y D2-D3 (-54.48 por ciento y -60.15 por ciento vs. 1553.97 por ciento y 0 por ciento); lo cual no ocurrió con la PCR. Se observó una débil correlación de la PCT con la PCR en el tiempo (r=0,46). La correlación de la PCT con APACHE II y MODS fue mayor que con la PCR (r=0.88 y 0.81 vs 0.60 y 0.53). Conclusiones: una disminución >30 por ciento entre D1-D2 y >50 por ciento entre D2-D3 de los niveles séricos de PCT se asocian a un pronóstico favorable. La PCT puede ser mas útil que la PCR en la monitorización diaria de la terapia en pacientes sépticos. La PCT tiene una mayor correlación que la PCR con el curso clínico del paciente séptico de acuerdo a los scores APACHE II y MODS

Aim: to observe the behavior of the procalcitonin serum levels during the clinical course of the sepsis in the local population. Methodology: prospective cohort study. Patients admitted to the intensive care unit of the Kennedy general hospital diagnosed with sepsis and without antibiotic treatment for more than 48 hours prior to admission were considered for the research. Serum levelsof procalcitonin (PCT) and C reactive protein (CRP) were obtained, and APACHE II and MODS scores were calculated at admission and daily until the resolution of the septic picture or death of the patient. Results: eight patients were included in the research. A higher percentage decrease in PCT was observed in survivors than non-survivors between D1-D2 and D2-D3 (-54.48 percent and -60.15 percent vs. 1553.97 percent and 0 percent); which did not occur with CRP. A weak correlation of PCT and CRP over time (r=0.46) was observed. The correlation of PCT with APACHE II and MODS was higher than with the CRP (r=0.88 and 0.81 vs 0.60 and 0.53). Conclusions: adecrease > 30 percent between D1-D2 and > 50 percent between D2-D3 serum levels of PCT are associated with a favorable prognosis. PCT can be of major use than CRP in terms of daily monitoring of the therapy in septic patients. PCT has a higher correlation with the clinical course of the septic patient than CRP according to the APACHE II and MODS scores.

Preparation and biodistribution study of [67]Ga-gallium calcitonin

In order to develop a radiolabeled calcitonin [CT] derivative for receptor imaging studies, CT was successively labeled with [67]G-gallium chloride. The best results of the conjugation were obtained by the addition of 0.5 ml of a CT nasal pharmaceutical solution [1100 IU] to a glass tube pre-coated with DTPA-dianhydride [0.01 mg] at 25 [degree sign] with continuous mild stirring for 30 min. after solid phase purification of the radiolabeled hormone, instant thin layer chromatography [ITLC] showed radiochemical purity of higher than 95% at optimized conditions [specific activity = 67-134 KBq/IU, labeling efficiency 70%]. [67]Ga-DTPA-CT mainly accumulates in the liver. Preliminary in vivo studies [ID/g%] in male wild-type rats showed significant liver uptake of the tracer after 24 hours. [67]Ga-DTPA-CT can be a suitable probe for biodistribution study of CT receptors in various physiological as well as neoplastic lesions with over-expressed calcitonin receptors

Expression of adrenomedullin and its receptor mRNA in the tissues of normal adrenal medulla and pheochromocytoma / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the expression of human adrenomedullin (ADM) and its receptor-receptor activity modifying protein 2/calcitonin receptor-like receptor (RAMP2/CRLR) mRNA in the tissues of normal adrenal medulla and pheochromocytoma.</p><p><b>METHODS</b>Total RNA was extracted from normal adrenal medulla and pheochromocytomas. The expression of ADM and RAMP2/CRLR mRNA were studied by reverse transcription-polymerase chain reaction. The ratios of ADM/GAPDH, RAMP2/ GAPDH, CRLR/GAPDH were used to evaluate the expression levels of ADM, RAMP2 and CRLR mRNA.</p><p><b>RESULTS</b>Expressions of ADM and its receptor- RAMP2/CRLR mRNA were detected in normal adrenal medulla and pheochromocytoma tissues. ADM/GAPDH were 0.48+/-0.09 and 0.75+/-0.24, RAMP2/ GAPDH 0.79+/-0.12 and 1.29+/-0.30, CRLR/GAPDH 0.40+/-0.08 and 0.87+/-0.22 in normal adrenal medulla and pheochromocytomas, respectively (P < 0.05).</p><p><b>CONCLUSION</b>ADM exerts a possible autocrine or paracrine effect in the adrenal. ADM may be involved in the pathogenesis of pheochromocytoma.</p>

The relationship between calcitonin receptor gene AluI polymorphism, bone mineral density and bone responsiveness to hormone replacement therapy in postmenopausal Korean women / 대한산부인과학회지

OBJECTIVE: To evaluate the relationship between the calcitonin receptor (CTR) gene AluI polymorphism, serum calcitonin levels, bone mineral density (BMD) and bone responsiveness to hormone replacement therapy (HRT). METHODS: The CTR AluI polymorphism were determined by restriction fragment length polymorphism (RFLP) in 433 postmenopausal Korean women. Among these women, 306 women received sequential HRT for 1 year. Serum bone alkaline phosphatase, CrossLaps, osteocalcin and calcitonin levels were measured by immunoassay and BMD at the lumbar spine and proximal femur by dual energy X-ray absorptiometry before and after HRT of 1 year. RESULTS: The CTR genotype frequencies were 81.3% for CC, 17.5% for CT, and 1.2% for TT. No significant differences in BMD at the lumbar spine and proximal femur and their annual percentage changes after HRT were noted among CTR genotypes. There were no significant differences in the levels of calcitonin or bone turnover markers and their 6 month percentage changes after HRT among CTR genotypes. The CTR genotypes were not distributed differently between HRT-responders and HRT-nonresponders (women who lose more than 3% of bone mass per year) or between women with normal BMD and women with low bone mass. CONCLUSION: The CTR AluI polymorphism is not associated with BMD and bone responsiveness to HRT in Korean women.

Investigation of the expression of calcitonin receptor mRNA in human osteoclasts on deciduous teeth / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression of calcitonin receptor mRNA in the osteoclasts of the resorbing deciduous teeth.</p><p><b>METHODS</b>After fixing the collected deciduous teeth, toluidine blue was performed and tartrate-resistant acid phosphatase (TRAP) staining was used to identify the osteoclasts on the resorbing surface of human deciduous teeth and in situ hybridization of calcitonin receptor mRNA to show its existence.</p><p><b>RESULTS</b>There were a number of TRAP positive osteoclasts on the root surface which showed the expression of calcitonin receptor mRNA.</p><p><b>CONCLUSION</b>On the resorbing surface of human deciduous teeth there are osteoclasts that express calcitonin receptor mRNA, so it is feasible to use this kind of osteoclast to test the effect of external factors on the expression of CTR mRNA.</p>

Increased atria expression of receptor activity-modifying proteins in heart failure patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Receptor activity-modifying proteins (RAMPs) determine the ligand specificity of the calcitonin receptor-like receptor (CRLR); co-expression of RAMP1 and CRLR results in a calcitonin gene related peptide (CGRP) receptor, whereas the association of RAMP2 or RAMP3 with CRLR gives an adrenomedullin(ADM) receptor. As CGRP and ADM may play a beneficial role in heart failure, this study aimed at the question whether RAMPs mRNAs are changed in heart failure.</p><p><b>METHODS</b>Semi-quantitative reverse transcription-PCR (RT-PCR) was used to detect and quantify the mRNAs of RAMP1 and RAMP3 in the atria of heart failing patients.</p><p><b>RESULTS</b>It was found that the expressions of RAMP1, RAMP2 and RAMP3 mRNAs increased with the worsening of heart function, but the expressions of RAMP1 and RAMP2 mRNA decreased at level IV of heart failure.</p><p><b>CONCLUSION</b>The above results demonstrated in the atria of heart failure patients an up-regulation of CGRP receptor by an increase of RAMP1 in association with CRLR and an up-regulation of ADM receptor by an increase of RAMP2 expression in association with CRLR, thus suggesting that CGRP and ADM receptors be playing a functional role in compensating the chronic heart failure in human.</p>

Association of calcitonin receptor gene polymorphism with bone mineral density in Shanghai women / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the association of calcitonin receptor (CTR) gene polymorphism with bone mineral density (BMD) in premenopausal and postmenopausal women.</p><p><b>METHODS</b>CTR genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 184 premenopausal women and 199 postmenopausal women in Shanghai area. BMD at lumbar spine (L2-4) and femoral neck (FN) were measured by dual-energy X-ray absorptiometry (DEXA).</p><p><b>RESULTS</b>The distribution of CTR genotypes in 383 Shanghai women were CC genotype 83.8%, TC genotype 14.6%, TT genotype 1.6%, respectively. BMD at FN of CC genotype was significantly higher than TC and TT genotypes (P < 0.01) in postmenopausal women. But there was no difference in BMD of different CTR genotypes in premenopausal women. Multiple regression analysis showed that CTR genotypes were associated with FN BMD in postmenopausal women (P < 0.05).</p><p><b>CONCLUSIONS</b>The polymorphism of CTR gene was associated with BMD in postmenopausal women.</p>

Role of decisive markers in diagnosis and outcome of patients with septic shock

This work aimed to assess the potential role of procalcitonin [PCT] and polymorphonuclear [PMN] elastase enzyme in the early diagnosis and early prediction of prognosis in patients with sepsis and septic shock. Twenty patients with septic shock [16 males and 4 females, mean age 50.15 years] together with a second group comprising 10 patients [9 males, mean age 49.2 years] with systemic sepsis without shock were studied. A third group including 20 healthy volunteers matching with age and sex and served as controls. Serum PCT and PMN elastase enzyme levels were estimated on admission for both patients and control groups with other laboratory investigations and clinical parameters. A multivariate discriminate analysis was performed using PCT, PMN elastase enzyme, albumin, alpha-1-antitrypsin, alpha-2 macroglobulin and C-reactive protein [CRP] as independent parameters. The study concluded that serum PCT and PMN elastase enzyme are independent useful diagnostic markers for the early detection of systemic inflammatory response syndrome with or without shock. However, PCT has the advantage over the above mentioned parameters having significantly predictive accuracy of 80%. Procalcitonin, PMN elastase enzyme, alpha-1-antitrypsin, alpha-2-macroglobulin, CRP and albumin could be used for the early prediction of complications of sepsis patients with an overall predictive accuracy of 76.7%

Human Periodontal Ligament Fibroblasts Support the Osteoclastogenesis of RAW264.7 Cells / 대한치주과학회지

The fibroblasts are the principal cells in the periodontal ligament of periodontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLF enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to clarify the mechanism of PDLF-induced osteoclastogenesis and 2) whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptor activator nuclear factor kappa B (RANK), 2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the OAASTM plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing activity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture medium. The effects of PDLF on differentiation of RAW264.7 into the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fixed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1beta-treated, or lipopolysaccharide- treated and then fixed PDLF showed two-fold increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findings suggested that we can replace the primary hematopoietic preosteoclasts for RAW264.7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

The localization of adrenomedullin in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC).</p><p><b>METHODS</b>A monoclonal antibody against AM developed by our laboratory was used to detect the localization of AM protein in rat kidney tissue by avidin-biotin complex immunohistochemistry. The expressions of AM and its receptor CRLR mRNA on cultured glomerular epithelial cells (GEC) and MsC were investigated by Northern blot assay, and the possible effect of AM secreted by GEC on MsC proliferation was observed using [3H]thymidine incorporation as an index.</p><p><b>RESULTS</b>A specific monoclonal antibody against AM was succesfully developed. AM was immunohistochemically localized mainly in glomeruli (GEC and endothelial cells), some cortical proximal tubules, medullary collecting duct cells, interstitial cells, vascular smooth muscle cells and endothelial cells. Northern blot assay showed that AM mRNA was expressed only on cultured GEC, but not on MsC, however, AM receptor CRLR mRNA was only expressed on MsC. GEC conditioned medium containing AM can inhibit MsC growth and AM receptor blocker CGRP8-37 may partially decreased this inhibitory effect.</p><p><b>CONCLUSION</b>AM produced by GEC inhibits the proliferation of MsC, which suggests that AM as an important regulator is involved in glomerular normal physiological functions and pathologic processes.</p>

Calcitonina: fisiologia e deficiência / Calcitonin: physiology and defficiency

O desenvolvimento e o aprimoramento das técnicas de dosagem da calcitonina sérica (CT) (radioimunoensaio e ensaios imunométricos), os avanços da biologia molecular, abriram novas e interessantes perspectivas, permitindo amplicar o conhecimento acerca da CT, este hormônio tantas vezes dito "sem funçao". Nesta revisao, sao discutidos aspectos históricos, estruturais e moleculares da CT, os métodos de dosagem, a importância e utilidade dos testes provocativos, sobretudo no diagnóstico do câncer medular da tireóide. Além disso, sao abordados os principais aspectos fisiológicos conhecidos, bem como as condiçoes caracterizadas por deficiente reserva secretóia de CT como no hipotireoidismo congênito, na tireoidite crônica autoimune, em pacientes tireoidectomizados e em mulheres no período pós-menopausa.