Epitope identification of calcitonin gene-related peptide antibodies based on ELISA and NMR methods / 中国生物制品学杂志

@#Objective To develop an indirect ELISA-based peptide scanning method combined with nuclear magnetic resonance(NMR)technique for the epitope identification of calcitonin gene-related peptide(CGRP)antibodies.Methods The antigen binding activities of two antibodies(new CGRP antibody and control antibody)were determined by indirect ELISA using each truncated CGRP fragment as coating antigen,and the linear epitope was analyzed according to the EC50value of four-parameter curve. Two-dimensional hydrogen-nitrogen correlation(2D1H-15N HSQC)spectrum of CGRP were acquired by NMR technique,and the binding of antibodies to the arginine of CGRP were analyzed through the disturbance of the antibodies to CGRP signals. Specific arginine modifications were detected by liquid chromatography-mass spectrum(LCMS) and NMR technique,and two arginine resonances were assigned on CGRP by correlating the rank order of the modification rate.ResultsThe antigen binding activities of two antibodies with CGRP(1-37),CGRP(19-37)and CGRP(25-37)showed dose-response relationships,and were fitted with four-parameter equation. However,there were no significant antigen binding with CGRP(1-18),CGRP(19-24)and CGRP(25-37)without C-terminal amide. The linear epitopes of both antibodies were located at the C-terminal of CGRP. The resonances of arginine ε-NH in 2D1H-15N HSQC spectrum disappeared in the presence of the control antibody;and the resonances appeared in the presence of the new antibody. The arginine R11 and R18 of CGRP could bind to the control antibody,but not to the new antibody. The NMR assignment for the arginine resonances were made by correlating the relative ranking of the modification rate where signals A and B arose from R11 ε-NH and R18 ε-NH respectively.ConclusionIn this study,the linear and conformational epitopes of new CGRP antibody and control antibody were identified based on the methods of ELISA and NMR,which may provide a theoretical basis for the design of the candidate therapeutic CGRP antibodies.

Analgesic Effect of Intrathecal 2R， 6R-HNK on Neuropathic Pain in Female Mice / 中山大学学报(医学科学版)

ObjectiveTo investigate the analgesic action and mechanism of intrathecal 2R， 6R-hydroxynorketamine （2R， 6R-HNK） on spared nerve injury （SNI）-induced chronic neuropathic pain （CNP） in female mice. MethodsSNI was used to establish acute and chronic CNP models in female mice. The mice were randomly divided into different groups with administration of vehicle， 2R， 6R-HNK or S-ketamine （10 mg/kg intraperitoneal injection/i.p. or 7， 21 μmol/L intrathecal injection/i.t.） at 3 weeks after or 30 min/1 d before operation （n = 3 - 7 mice/group）. The curative or preventive effect of 2R， 6R-HNK was evaluated by mechanical paw withdrawal threshold （PWT） and the analgesic efficiency. Finally， immunofluorescence and RT-PCR of dorsal root ganglion （DRG） and spinal dorsal horn （SDH） were used to explore the possible mechanisms. ResultsCompared with vehicle， intrathecal injection of 2R， 6R-HNK largely reversed SNI-induced bilateral mechanical allodynia in a delayed-and-dose-dependent way. Among them， 21 μmol/L 2R， 6R-HNK reached its maximum analgesic efficiency （75.32±7.69） % at 2 d. Pre-intrathecal delivery of 2R， 6R-HNK also delayed the development of bilateral mechanical hypersensitivity 2 - 3 d induced by SNI. Mechanically， 2R， 6R-HNK reversed not only the abnormal excitability of neurons in bilateral DRG and superficial SDH， but also the upregulation of calcitonin gene-related peptide （CGRP） and brain-derived nerve growth factor （BDNF） in DRG. ConclusionIntrathecal administration of 2R， 6R-HNK exerts an analgesic effect against CNP， probably via suppressing abnormal neuronal excitability in ascending pain pathway as well as down-regulating CGRP and BDNF expression in DRG neurons.

Effect of moxibustion on colonic mucosal injury and expression of CGRP positive nerve fibers of distal colonic mucosa in ulcerative colitis mice / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion at "Zusanli" (ST 36) on distal, middle and proximal colonic mucosal injury and expression of calcitonin gene-related peptide (CGRP) positive nerve fibers of distal colonic mucosa in ulcerative colitis (UC) mice at different time points.@*METHODS@#A total of 51 C57BL/6N mice were randomized into a 7-day control group (@*RESULTS@#Mucosal injury can be observed in mice after modeling, displaying epithelial layer disappearance, abnormal crypt structure or crypt disappearance. Compared with the 7-day control group, colon length was shortened (@*CONCLUSION@#Moxibustion at "Zusanli" (ST 36) can reduce the expressions of positive nerve fibers of colonic mucosa and CGRP positive nerve fibers of distal colonic mucosa, thus, improve the colonic mucosal injury.

The role of calcitonin gene-related peptide in migraine prevention by botulinum toxin type A

@#Objectives: Calcitonin gene-related peptide (CGRP) is currently considered to be a major contributing factor in migraine headache. Botulinum toxin type A (BTXA) was found to be effective in migraine prevention. However, the mechanism of action in patients was unknown. Using injection as in clinical setting, the study aimed to determine whether BTXA could decrease the sensitization of the trigeminovascular nociceptive system through the reduction of CGRP action. Methods: Adult male Wistar rats were pretreated with normal saline solution or BTXA before KCl application to induce cortical spreading depression (CSD) or NaCl application as a control. Regional cerebral blood flow at parietal cortex was measured for 90 min after KCl or NaCl application. Tissues from trigeminal ganglion (TG) and trigeminal nucleus caudalis (TNC) were then collected for CGRP and c-Fos measurement respectively. Results: BTXA pretreatment significantly decreased the cumulative blood flow and number of hyperemic peaks induced by KCl. Numbers of CGRP positive cells at TG and c-Fos positive cells at TNC were also reduced by BTXA.Conclusion: BTXA pretreatment reduced CGRP production and release from the TG leading to lessen CSD production and persistent activation of TNC which played a major role in migraine headache.

Effects of Naoluo Xintong Decoction on vascular dementia rats' memory and learning, and hippocampal neuronal intracellular calcium concentration / 中成药

AIM To study the effects of Naoluo Xintong Decoction (NLXTD) on vascular dementia (VD)rats' memory and learning,and hippocampal neuronal intracellular calcium concentration.METHODS Rats were divided randomly into model group,NLXTD group (8.54 g/kg),Huperzia-A group (0.06 mg/kg) and sham group.They were made into vascular dementia rats by the improved bilateral carotid artery ligation method (2-VO)thereafter if necessary.After one-month corresponding intragastric administration,the rats were ethologically evaluated by the Morris water maze experiment;their fluorescence intensity of hippocampal neuronal intracellular calcium concentration was determined by flow cytometry,and the expression levels of calcitonin gene related peptide (CGRP) and its hippocampus receptor were detected by enzyme-linked immunosorbent assay (ELISA).RESULTS Compared with the model group,rats in NLXTD group displayed overall superiority to those of the model group in terms of significantly shortened time of escape latency (P ＜0.01),significantly increased number through the platform and the times in the fourth quadrant (P ＜ 0.01),a lower fluorescence intensity indicating a lower hippocampal neuronal intracellular calcium concentration (P ＜ 0.05).CONCLUSION The significant improvement of memory and learning observed among VD rats' due to NLXTD intervention may be attributable to its efficacy in reducing the hippocampal neuronal intracellular calcium concentration by enhancing the expression levels of CGRP and its receptor.

The effects of intra-articular resiniferatoxin on monosodium iodoacetate-induced osteoarthritic pain in rats

This study was performed to investigate whether an intra-articular injection of transient receptor potential vanilloid 1 (TRPV1) receptor agonist, resiniferatoxin (RTX) would alleviate behavioral signs of arthritic pain in a rat model of osteoarthritis (OA). We also sought to determine the effect of RTX treatment on calcitonin gene-related peptide (CGRP) expression in the spinal cord. Knee joint inflammation was induced by intra-articular injection of monosodium iodoacetate (MIA, 8 mg/50 microl) and weight bearing percentage on right and left hindpaws during walking, paw withdrawal threshold to mechanical stimulation, and paw withdrawal latency to heat were measured to evaluate pain behavior. Intra-articular administration of RTX (0.03, 0.003 and 0.0003%) at 2 weeks after the induction of knee joint inflammation significantly improved reduction of weight bearing on the ipsilateral hindlimb and increased paw withdrawal sensitivity to mechanical and heat stimuli. The reduction of pain behavior persisted for 3~10 days according to each behavioral test. The MIA-induced increase in CGRP immunoreactivity in the spinal cord was decreased by RTX treatment in a dose-dependent manner. The present study demonstrated that a single intra-articular administration of RTX reduced pain behaviors for a relatively long time in an experimental model of OA and could normalize OA-associated changes in peptide expression in the spinal cord.

Experimental study on the effect of RAMP1 -siRNA on CGRP-induced proliferation of MG-63 cells / 实用口腔医学杂志

Objective:To investigate the effect of blocking the expression of receptor activity modifying protein 1 (RAMP1 )on calcito-nin gene-related peptide(CGRP)-induced MG-63 cell proliferation.Methods:RAMP1 siRNA was synthesized and screened by tran-scription in vitro.The subcultured MG-63 cells were divided into the following groups:RAMP1 siRNA interference group,empty vector group and blank control group.The mRNA expression and the membrane distribution changes of the calcitonin receptor-like receptor (CRLR)and the receptor component protein (RCP)in MG-63 cells were examined by real-time PCR and immunofluorescence method respectively.Results:RAMP1 and CRLR mRNA and the fluorescence intensity of MG-63 cells decreased after transfection by RAMP1 siRNA(P <0.05).In RAMP1 interference group,the expression of RCP mRNA and the fluorescence intensity were higher than those in the other two groups(P <0.05).After RAMP1 siRNA interference,the proliferation of MG-63 cells was inhibited(P <0.05). Conclusion:RAMP1 siRNA transfection may reduce CRLR expression and inhibite the proliferation of MG-63 cell.

Function of calcitonin gene-related peptides in chronic morphine tolerance and hyperalgesia / 中国肿瘤临床

Pain is a common symptom in patients with terminal cancer and is the main factor that affects their quality of life. Morphine is commonly used for the treatment of acute and chronic pain, but the long-time application of morphine results in morphine tolerance and hyperalgesia, which restrict the clinical applications of the anesthetic. In this review, pertinent studies on morphine over recent years were summarized and analyzed, and the mechanism of morphine tolerance and hyperalgesia were discussed, specifically on the function of calcitonin gene-related peptide (CGRP) in clinical practice. The authors analyzed the relationship between the plastic changes in the nervous system and chronic morphine application in terms of CGRP up-regulation based on its molecular biology charac-teristics and distribution, the relationship between CGRP and chronic application of morphine, and between CGRP and hyperalgesia. The effect of CGRP up-regulation on the nociceptive system and the relationship between CGRP up-regulation and the formation of hy-peralgesia were also analyzed. We discussed the sensitized effect of CGRP up-regulation on the nociceptive system, which promotes morphine tolerance and hyperalgesia. This study provides a framework for treating morphine tolerance and can be used as a guide for further research on the topic.

Recovery of Motor Function and Changes of Anterior Horn Cells after Spinal Cord Injury in Rats / 中国康复理论与实践

@#Objective To explore the restoration of motor function and the expression of calcitonin gene-related peptide(CGRP)and acetylcholine esterase(AChE)in the anterior horn motoneurons after different types of spinal cord injury.Methods 60 adult female Wistar rats were randomly assigned to 3 groups:sham group,completely transection group and contusion group.Average combined scores(ACOS)were applied to assess the motor function at various time after the surgery.The content of AChE in the anterior horn of L2-L4 was detected with Karnovsky-Roots staining and the expression of CGRP was then determined with immunohistochemistry.Results The scores of ACOS were much higher in the contusion group than in the transection group at each time point examined.The content of both AChE and CGRP significantly decreased after either type of spinal cord injury.However,their activity gradually recovered to the normal level in the contusion group,but not in the transection group.Moreover,the changes of CGRP occurred earlier than those of AChE.Conclusion There is strong relationship between the motor function recovery and the functional state of anterior horn cells.CGRP or AChE may play an important role in the functional recovery of locomotion after spinal cord injury in rats.

Changes of Calcitonin Gene-related Peptide and Acetylcholine Esterase in Motor End Plates after Spinal Cord Injury in Adult Rats / 中国康复理论与实践

@#Objective To explore the degeneration of motor end plates (MEP) by observing the expression of calcitonin gene-relative peptide (CGRP) and acetylcholine esterase (AChE) in the MEP after different types of spinal cord injury. Methods 60 adult female Wistar rats were randomly assigned to 3 groups: sham group, completely transection group and contusion group. The content of AChE in the MEP was detected with Karnovsky-Roots staining and the expression of CGRP was then determined with immunohistochemistry. Results The content of both AChE and CGRP significantly decreased after either type of spinal cord injury. However, their activity gradually recovered to the normal level in the contusion group, but not in the transection group. Moreover, the changes of CGRP occurred earlier than those of AChE. Conclusion The motor end plate degenerates differently after different kinds of spinal cord injury in adult rat, CGRP and AChE are related to the degeneration of MEP.

Effect of scalp point penetration combined with rehabilitation training on endothelin and calcitonin gene-related peptide in plasma of patients with acute cerebral hemorrage / 中国康复理论与实践

@#ObjectiveTo investigate the effect of scalp point penetration combined with rehabilitation training on content of endothelin (ET) and calcitonin gene-related peptide (CGRP) in plasma of patients with acute cerebral hemorrage (ACH).Methods90 ACH patients were randomly divided into the scalp point penetration combined with rehabilitation group (group A), the rehabilitation group (group B) and control group (group C) with 30 cases in each group. The radioimmunoassay was adopted to determinate the content of ET and CGRP in plasma of patients.ResultsAfter treatment, contents of ET and CGRP of all patients were decreased significantly (P<0.01), but group A and group B had a significant difference compared with the group C (P<0.01), and there was also a significant difference between the group A and group B.ConclusionThe scalp point penetration combined with rehabilitation training can regulate the content of ET and CGRP in plasma and make them under a dynamic balance.

Regulation of pulpal microcirculation by calcitonin gene-related peptide / 대한치과보존학회지

The purpose of this study was to investigate the function of calcitonin gene-related peptide (CGRP) in regulatory mechanism of pulpal microcirculation with the aim of elucidating neurogenic inflammation. Experiments were performed on twelve cats under general anesthesia. CGRP was administered through the femoral vein to see the systemic influence and through the external carotid artery to see the local effect. Sympathetic nerve to the dental pulp was stimulated electrically and pulpal blood flow (PBF) was measured with a laser Doppler flowmeter on the canine teeth to the drug administration. The paired variables of control and experimental data were compared by paired t-test and differences with p < 0.05 were considered statistically significant. Systemic administration of CGRP (0.3 microg/kg) exerted decreases in systemic blood pressure and caused changes in PBF with an initial increase followed by decrease and a more marked second increase and decrease. Close intra-arterial (i.a.) injection of CGRP (0.03 microg/kg) resulted in slight PBF increase. The effect of CGRP resulted in no significant increase in PBF in the presence of CGRP8-37. The electrical stimulation of the sympathetic nerve alone resulted in PBF decreases. The i.a. administration of CGRP following the electrical stimulation of the sympathetic nerve compensated the decreased PBF. Therefore, CGRP effectively blocked the sympathetic nerve stimulation-induced PBF decrease. Results of the present study have provided evidences that even though the local vasodilatory function of CGRP are weak, CGRP is effectively involved in blocking the vasoconstriction caused by sympathetic nerve stimulation in the feline dental pulp.

Degeneration of primary afferent terminals following brachial plexus extensive avulsion injury in rats / Degeneración de los terminales aferentes primarios de rata luego de lesión extensa por avulsión del plexo braquial

Important breakthroughs in the understanding regeneration failure in an injured CNS have been made by studies of primary afferent neurons. Dorsal rhizotomy has provided an experimental model of brachial plexus (BP) avulsion. This is an injury in which the central branches of primary afferents are disrupted at their point of entry into the spinal cord, bringing motor and sensory dysfunction to the upper limbs. In the present work, the central axonal organization of primary afferents was examined in control (without lesion) adult Wistar rats and in rats subjected to a C3-T3 rhizotomy. Specific sensory axon subtypes were recognized by application of antibodies to the calcitonin gene-related peptide (CGRP), the P2X3 purinoreceptor, the low-affinity p75-neurotrophin receptor and the retrograde tracer cholera toxin subunit beta (TCbeta ). Other subtypes weres labeled with the lectin Griffonia simplicifolia IB4. Using immunohistochemistry and high resolution light microscopy, brachial plexus rhizotomy in adult rats has proven a reliable model for several neural deficits in humans. This lesion produced different degrees of terminal degeneration in the several types of primary afferents which define sub-populations of sensitive neurons. Between the C6 and C8 levels of the spinal cord,,deafferentation was partial for peptidergic GCRP-positive fibers, in contrast with elimination of non peptidergic and myelinated fibers. Dorsal rhizotomy has provided an adequate experimental model to study sensory alterations such as acute pain and allodynia as well as factors that affect regeneration into the CNS., Therefore, the differential deafferentation response must be considered inr the evaluation of therapies for nociception (pain) and regeneration for brachial plexus avulsion. The anatomical diffierences among the primary afferent subtypes also affect their roles in normal and damaged conditions.

El uso de las neuronas sensoriales primarias ha aportado avances en el entendimiento de las razones por las cuales falla la regeneración cuando el sistema nervioso central (SNC) es dañado. La rizotomía dorsal se puede usar como un modelo experimental de las lesiones por avulsión del plexo braquial, una lesión en la cual son desprendidas, en su punto de entrada en la médula espinal, las ramas centrales de los aferentes primarios causando una disfunción motora y sensorial grave e irreversible del miembro superior. En el presente trabajo, se examinó la organización central de los aferentes primarios en ratas Wistar adultas. Éstas fueron divididas en controles normales no lesionados y en animales rizotomizados entre los niveles cervical 3 y torácico 3 (C3-T3). Se estudió la deaferentación de los subtipos de axones sensoriales utilizando anticuerpos específicos contra el péptido relacionado con el gen de la calcitonina (CGRP), el receptor purinérgico (P2X3), el receptor de baja afinidad p75 para el factor de crecimiento nervioso (NGF) y contra la subunidad ®de la toxina de cólera (TCbeta ). Otro subtipo fue marcado con la lectina Griffonia simplicifolia IB4. La inmunohistoquímica y la microscopía óptica de alta resolución demostraron que el modelo animal de rizotomía completa del plexo braquial reproduce diversos déficit observados en las lesiones humanas. Esta lesión produce diferentes grados de degeneración terminal entre los diversos tipos de aferentes primarios que definen subpoblaciones de neuronas sensoriales. En los niveles de la médula espinal estudiados (entre C6 y C8), la deaferentación fue parcial para las fibras peptidérgicas GCRPpositivas, en contraste con la eliminación de las fibras no peptidérgicas y las mielinizadas. La rizotomía dorsal es un modelo experimental apropiado para estudiar las alteraciones sensoriales como el dolor agudo y la alodinia, así como los factores que podrían afectar la regeneración en el SNC. Por tanto, la respuesta de deaferentacion diferencial debe ser tenida en cuenta para la evaluación de terapias antinociceptivas y regenerativas tras la avulsión del plexo braquial. Se discute la anatomía de los subtipos de aferentes primarios y su papel en condiciones normales y después de la lesión.

Jiuqiang Naoliqing enhancing the expression of CGRP and Synapsin Ⅰ in brain of spontaneous hypertension rats / 中国康复理论与实践

@#ObjectiveTo study the influence of Jiuqiang Naoliqing (JNQ) on the expression of calcitonin gene related peptide(CGRP)and Synapsin Ⅰ in brain of the spontaneous hypertension rats (SHR). MethodsThe rats were randomly divided into 4 groups: Wistar group, SHR group, lower dose of JNQ treated SHR group and higher dose of JNQ treated SHR group. The expression of CGRP and Synapsin Ⅰ in the dentate gyrus, CA1 subfield of hippocampus and cortex were determined by immunohistochemistry after treatment for 3 weeks. ResultsCompared with the Wistar group, the expression of CGRP and Synapsin Ⅰ in the dentate gyrus, CA1 subfield of hippocampus and cortex of SHR group significantly decreased. The treatment with lower dose of JNQ significantly enhanced the expression of CGRP in cortex(P<0.05 vs SHR).The treatment with higher dose of JNQ significantly enhanced not only the expression of CGRP in the dentate gyrus, CA1 subfield of hippocampus and cortex, but also that of Synapsin Ⅰ in the CA1 subfield of hippocampus selectively in comparison with SHR group. ConclusionJNQ may improve the micro circulation in brain by up regulating the expression of CGRP and enhance the modulating function of central nervous system by up regulating the expression of Synapsin Ⅰ in spontaneous hypertension rats.

Calcitonin Gene-related Peptide Immunoreactivity in the Muscle Layer of Small Intestine; Its Action on Interstitial Cell / 체질인류학회지

In addition to the central and the peripheral nervous system, calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) has been identified throughout the enteric nervous system. Several functions of the CGRP in gastrointestinal (G-I) tract has been identified, but the effect of CGRP on G-I motility is unclear. The distribution of calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) in the murine small bowel were studied by using immunohistochemistry, also analyzed functionally by using electrophysiological method. Immunohistochemical studies demonstrated that CGRP-LI is localized in both nerve fibers and myenteric ganglion cells in the whole-mount preparation of murine small intestine. Double labelling with CGRP and c-kit investigated by confocal microscope was shown that CGRP-LI enteric nerve fiber surrounded the c-kit positive interstitial cells of Cajal (ICC). Electrophysiological finding revealed that treatment of CGRP inhibited electrical activity on culture ICC. Our results suggest a CGRP innervation of murine small bowel ICC. The released CGRP from enteric nerve terminals may induce relaxation of small bowel through the inhibition of ICC.

Effects of tegaserod on visceral sensitivity and expression of SP and CGRP in the rat colon / 西安交通大学学报(医学版)

Objective To investigate the effects of tegaserod on visceral sensitivity and explore the regulating mechanism.Methods Forty-two male Spragre-Dawley rats,which were induced colonic inflammation by intraluminal administration of trinitrobenzenesulfonic acid(TNBS),were randomly divided into eight groups.In the three colorectal distention(CRD) treated groups(n=6),abdominal contractions were recorded after 3,7 and 14 days of intra-gastric administration of tegaserod 2mg/kg d.In the three CRD control groups(n=4),abdominal contractions were recorded after 3,7 and 14 days of intra-gastric injection of saline 2.0mL/d.In immunohistochemistry(IH) treated group(n=6) and IH control group(n=6),samples of colon were removed and processed for SP and CGRP immunohistochemistry after 7 days of intra-gastric administration of tegaserod and saline,respectively.Results Abdominal contractions induced by colonic distention decreased significantly at 1.2mL and 1.6mL distention volume after 3 days of tegaserod administration(P

The role of calcitonin gene-related peptide in hypertension / 中国药理学通报

The sensory merves innervate blood vessels all over the body and participate in the regulation of cardiovascular function both in the normal state and in the pathophysiology of hypertension through the actions of potent vasodilator neuropeptides Calcitonin gene related peptide, the predominant neurotransmitter in the capsaicin sensitive sensory nerves, plays an important role in the initiation and progression of hypertension via the alterations in its synthesis and release.

The Distribution of CGRP Containing Cells Innervating the Calcaneal Tendon in Rat Dorsal Root Ganglia / 대한해부학회지

This study was performed to investigate origins of the dorsal root ganglion cells containing calcitonin gene -related peptide (CGRP) which innervate the calcaneal tendon in the rat. We used the horseradish peroxidase (HRP) or fluoro -gold (FG) to trace retrogradely somatic afferents in dorsal root ganglion cells after unilateral injections into the rat calcaneal tendon. HRP or fluoro -gold labeled DRG cells for the calcaneal tendon were seen generaaly in lumbosacral (L1 to S1) DRGs ipsilaterally. In lumbosacral DRGs, the largest number of labeled cells were found in the L6 DRG. Many DRG cell bodies contained the CGRP throughout the L1~S1. A plenty of HRP -or FG -labeled cells innervating the calcaneal tendon were also identified to contain the CGRP in L1~S1 DRGs. These FG +/- CGRP DRG cells innervating the calcaneal tendon were primarily found in the L6 DRG. These results suggest that the main sensory DRG for the calcaneal tendon is the L6. This fact may be available in diagnosis and treatment of neurogenic pain in the calcaneal tendon.

The effect of calcitonin gene-related peptide on the expression of osteoprotege rin and receptor activator of nuclear factor-?B ligand mRNA in rabbit osteobla sts / 实用口腔医学杂志

Objective:To elucidate the effects of calcitonin gene-re la ted peptide(CGRP) on the gene expression of osteoprotegerin(OPG) and receptor ac tivator of nuclear factor-?B ligand(RANKL) in rabbit osteoblasts. Meth ods:Osteoblasts were cultured in media containing 10 -10～10 -7 mol/L of CGRP. After 24-hour incubation,semi-quanitative RT-PCR was perfor med to detect the expression of OPG and RANKL mRNA,and with ?-actin mRNA as th e internal control. Results:CGRP increased the mRNA expressio n of OPG with the maximal effect at the concentration of 10 -8～10 -7 mol/L. CGPR downgulated the mRNA expression of RANKL dose-dependantly.C onclusion:CGRP may regulate the activities of osteoclasts by regulating gene expression of OPG/RANKL.

Study of peripheral and central mechanism of visceral hypersensitivity in non-erosive reflux disease patients / 中国实用内科杂志

Objective The aim of this study was to evaluate and to characterize the visceral hypersensitivity in non-erosive reflux disease(NERD)through the detection of esophageal distension-cerebral evoked potentials(ED-CEP)and calcitonin gene related peptide(CGRP)and substance P(SP)levels in esophageal mucosa.Methods NERD patients(N 26)and 12 controls were enrolled in this study from Oct.2004 to Mar.2005.Mechanical distention stimulation was performed using the balloon-affixed and polyvinyl multilumen catheter.The ED- CEP was recorded with a muti-channel international 10~20 system of electroencephalograph.The esophageal specimens were immune-histologically evaluated for CGRP/SP-stain positive contents.Results Esophageal distention may evoke recognizable and reproducible muti-peak CEP.CEP morphology of NERD patients was characterized by randomly distributed patterns,and the peak latencies for N1,P1,and N2 were significantly shorter compared with control group(P