Calcitonin gene-related peptide (CGRP) play a key role in some physiological and pathological progresses. The latest studies indicate that CGRP might involve in some disease progress and has a close relation with wound healing. It is significant to further investigate and then apply it to clinical diagnosis and therapy as well as forensic pathology.
Animals ; Calcitonin Gene-Related Peptide/physiology* ; Forensic Medicine ; Humans ; Receptors, Calcitonin Gene-Related Peptide/physiology* ; Wound Healing

PURPOSE: We attempted to investigate whether calcitonin gene-related peptide binding to receptors in the gubernaculum is different between normal and flutamide-treated rats or pups and whether calcitonin gene-related peptide(CGRP) binding is androgen dependent. MATERIALS AND METHODS: Timed pregnant Sprague Dawley rats were injected with flutamide or vehicle alone once daily on gestational days 15-19. Weight, anogenital distance and distance from testicle to symphisis pubis of pups of gestational day 20 and rats of neonatal day 1 and 7 were measured. Gubernacular sections from rats of neonatal day 7 were incubated with [125I]human CGRP with various concentrations of unlabeled hCGRP, and those from pups of gestational day 20 and rats of neonatal day 1 were incubated only with [125I]human CGRP. After exposure of gubernacular sections to imaging plate (BAS 2500), the images from the plate were quantified by computerized densitometry (TINA). RESULTS: Weight and anogenital distance of flutamide-treated pups or neonatal rats were significantly smaller and shorter than those of normal pups or neonatal rats (P< 0.01). Though the distance from testicle to symphisis pubis was not significantly different between normal and flutamide-treated pups or neonatal 1 day rats, that of flutamide-treated neonatal 7 day rats was significantly longer than that of normal neonatal 7 day rats (P< 0.01). The total binding counts of [125I]human CGRP on gubernacular sections of normal pups, neonatal 1 day rats and neonatal 7 day rats were 56.3 +/- 24.74, 68.2 +/- 24.90, 78.4 +/- 17.25 (dpm/mg polymer), respectively, and those of flutamide- treated pups, neonatal 1 day rats and neonatal 7 day rats were 43.7 +/- 12.54, 35.1 +/- 8.25, 57.5 +/- 16.27, respectively. There were significant differences between normal and flutamide-treated neonatal 1 day and 7 day rats (P< 0.01). The binding in normal rats was consistently increased from gestational day 20 to neonatal day 7, and it showed weak correlation (r = 0.398, P< 0.05). The binding analysis showed that concentrations of CGRP receptors were 20.0+/- 4.78 amol/mg polymer, 13.3 +/- 3.87 amol/mg polymer for normal and flutamide treated neonatal 7 day rats, respectively, and there was significant difference between normal and flutamide-treated rats (P< 0.01). However there was no significant difference in the dissociation constant between 2 models. The images from the plate in flutamide-treated neonatal 7 day rats looked smaller than those in normal 7 day rats. CONCLUSIONS: These results suggest that the inguinoscrotal descent of testicle occurs after the gubernacular eversion, CGRP binding in the gubernaculum is androgen dependent, and androgen may not influence CGRP release from genitofemoral nerve because of down regulation of CGRP receptor by antiandrogen. However, the role of CGRP in testicular descent is still obscure and the mechanism of down regulation of CGRP receptor by antiandrogen needs further investigation
Animals ; Calcitonin Gene-Related Peptide* ; Calcitonin* ; Cryptorchidism ; Densitometry ; Down-Regulation ; Flutamide ; Male ; Polymers ; Rats* ; Rats, Sprague-Dawley ; Receptors, Calcitonin Gene-Related Peptide ; Testis

BACKGROUND: Neuroimmunocutaneous system alteration can be responsible for the induction and maintenance of the inflammatory process of psoriasis. OBJECTIVE: This study was carried out to examine the expression of Substance P (SP), calcitonin gene-related peptide (CGRP), somatostatin (SOM), neutral endopeptidase (NEP), SP receptor, and CGRP receptor in psoriatic lesions. METHODS: A skin biopsy was obtained from 10 psoriatic patients and 10 normal control subjects. Confocal laser scanning microscopy was performed. RESULTS: The SP and CGRP receptors consistently increased in psoriatic lesions, compared to the normal controls. CONCLUSION: The increased expression of neuropeptides and their receptors may be involved in the pathogenesis of psoriasis.
Biopsy ; Calcitonin Gene-Related Peptide ; Humans ; Microscopy, Confocal ; Neprilysin ; Neuropeptides* ; Psoriasis ; Receptors, Calcitonin Gene-Related Peptide ; Receptors, Neuropeptide ; Skin ; Somatostatin ; Substance P

OBJECTIVE: To investigate the clinical value of expression level of interleukin-2 receptor (IL-2R) and interleukin-8 (IL-8) in the fever patients with hematological malignancies. METHODS: A total of 121 inpatients in the First Affiliated Hospital of Anhui Medical University from April 2018 to October 2019 were enrolled in this study. The patients were separated into infection group (61 cases) and non-infection group (60 cases). In the meantime, 40 healthy people without fever or infection in the hospital for physical examination were set as matched group. C-reactive protein (CRP), procalcitonin (PCT), and cytokines were detected in all the patients with fever after admission and infection control. While, blood samples were taken from healthy people during physical examination. RESULTS: The expression levels of IL-2R in infection group were higher than those in the control group (P<0.001), and the level of serum IL-2R in infection group was also higher than that in the non-infection group (P<0.05). Based on Spearman analysis, in patients with malignant hematologic disease, serum IL-2R level was positively correlated with CRP (r=0.557, P<0.001) and IL-8 (r=0.479, P<0.001), and IL-8 level was positively correlated with CRP (r=0.318, P<0.001). Compared with the non-infection group, the area under the curve (AUC) for the level of CRP, PCT, and IL-2R of the infection group was 0.714 (95%CI: 0.623-0.806), 0.765 (95%CI: 0.680-0.851), and 0.761 (95%CI: 0.686-0.836), the sensitivity was 0.705, 0.852, and 0.705, and the specificity was 0.717, 0.70, and 0.60, respectively. While, AUC of CRP+PCT, CRP+IL-2R, PCT+IL-2R, and CRP+PCT+IL-2R was 0.789 (95%CI: 0.712-0.866), 0.702 (95%CI: 0.623-0.782), 0.757 (95%CI: 0.677-0.838), and 0.789 (95%CI: 0.712-0.866), the sensitivity was 0.738, 0.934, 0.705, and 0.738, and the specificity was 0.840, 0.470, 0.810, and 0.840, respectively. CONCLUSION CRP, PCT, IL-2R, and IL-8 are useful parameters for diagnosis of the infectious fever in patients with hematological malignancies, which provides the basis of initial diagnosis and rational use of antibioties for clinician.
Biomarkers ; C-Reactive Protein ; Calcitonin ; Calcitonin Gene-Related Peptide ; Hematologic Neoplasms ; Humans ; Interleukin-8 ; Protein Precursors ; Receptors, Interleukin-2 ; Sepsis

To explore the role of intrapulmonary neuropeptides in the development of airway hyperresponsiveness, we established an animal model of airway hyperresponsiveness (AHR) in rabbits by using ozone exposure. With the model, after test of the mechanics of respiration and bronchoalveolar lavage assay, the levels of vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP) in the lungs were determined by radioimmunoassay, and the expression of mRNA coding receptors of these two neuropeptides was evaluated by reverse transcriptional-polymerase chain reaction (RT-PCR). At the same time, the distribution of VIP receptor-1 (VIPR1) and CGRP receptor-1 (CGRPR1) in lung tissues and its time-course were examined by in situ hybridization. The results showed: (1) in ozone-stressing groups, airway resistance increased significantly and typical inflammatory pathological changes were observed in pulmonary tissue slides, including neutrophil and eosinophil infiltration, mucus exudation and bronchial epithelial cells (BECs) shedding; (2) with elongation of ozone exposure, the levels of VIP and CGRP in the lungs increased at first, reaching a peak on d 2 to 4, then decreased slowly, and CGRP peaked somewhat earlier than VIP; (3) mRNA expression of the two neuropeptide receptors in the lungs changed in a similar manner like VIP and CGRP, but the high level of mRNA expression of VIPR1 lasted longer than that of CGRPR1; and (4) in situ hybridization for neuropeptide receptors demonstrated that, in unstressed control, VIPR1 and CGRPR1 positive cells appeared in the airway epithelium, pulmonary interstitial and focal areas of airway and vascular smooth muscles. With the elongation of ozone exposure, hybridization stained deeper and the majority of positive cells were located around the vessels and bronchus except a few in the alveoli. At 8 d, only a small number of positive cells were seen in the lungs. From the results, it is concluded that ozone-stressing can induce the development of AHR, in which VIP and CGRP may play important roles. That implies, through binding to CGRPR1, CGRP stimulates an early inflammation response which contributes in cleaning up of irritants, while VIP exerts a later dampening of pulmonary inflammation response. These two neuropeptides may play sequential and complementary roles in the development of AHR.
Animals ; Bronchi ; pathology ; Bronchial Hyperreactivity ; chemically induced ; metabolism ; Bronchoalveolar Lavage Fluid ; Calcitonin Gene-Related Peptide ; metabolism ; Epithelium ; metabolism ; Lung ; metabolism ; Ozone ; Rabbits ; Receptors, Calcitonin Gene-Related Peptide ; metabolism ; Receptors, Vasoactive Intestinal Peptide ; metabolism ; Vasoactive Intestinal Peptide ; metabolism

AIMTo study the effect of CGRP receptor antagonist CGRP8-37 on nociceptive response and expression of nitric oxide synthase (NOS) and content of nitric oxide (NO) in the dorsal horn of the spinal cord of rats during formalin-induced inflammatory pain.

METHODSUsing formalin injection into right hind paw induced inflammatory pain. Counting the times of flinching reflex was used to observe the degree of spontaneous pain. NADPH-d histochemistry was used to observe the changes of NOS expression. The content of NO was observed by measuring the contents of nitrate/nitrite (NO3- / NO2-).

RESULTSspontaneous pain behavioral was elicited by formalin injection. The NOS expression and NO content significantly increased in the spinal cord at 24 h after formalin injection. Intrathecal injection of CGRP8-37 could significantly inhibit the response of spontaneous pain and the increases of NOS expression and NO content induced by formalin injection.

CONCLUSIONThe activation of CGRP receptors enhances NOS expression and NO production in the dorsal horn of the spinal cord during formalin-induced inflammatory pain.

Animals ; Calcitonin Gene-Related Peptide ; pharmacology ; Formaldehyde ; adverse effects ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Pain ; chemically induced ; metabolism ; Peptide Fragments ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitonin Gene-Related Peptide ; antagonists & inhibitors ; Spinal Cord ; drug effects ; metabolism

Women often complain gut symptoms during pregnancy and the luteal phase of the menstrual cycle. To investigate the relationship between ovarian steroids and the abnormal gut motility and sensitivity, the expressions of cholecystokinin (CCK), calcitonin gene-related peptide (CGRP) and their receptors in stomach were studied in ovariectomized rats. Blood samples were collected for estradiol (E(2)), progesterone (P(4)), CCK and CGRP radioimmunoassay. Expression of CCK(A) receptor in fundus was assessed by Western blot and CGRP receptor was determined by (125)I-CGRP radioligand binding assay (RBA). The replacement therapy with estradiol benzoate (EB) could dose-dependently increase the plasma CCK level and the expression of gastric CCK(A) receptor (P<0.05 respectively). P(4) replacement therapy could stimulate the release of CGRP and increase the binding sites of CGRP receptors in stomach (P<0.05 respectively). The combined effect of EB and P(4) was to stimulate the release of CCK and CGRP, and to increase the expressions of gastric CCK(A) and CGRP receptors. These results indicate that EB could inhibit gastric emptying by increasing CCK secretion and CCK(A) receptor expression in ovariectomized rats. P(4) could increase gut sensitivity by up-regulating the release of CGRP and the activity of CGRP receptor. It could be deduced from these observations that CCK(A) and CGRP receptor antagonists could be used for female patients who suffer from gastrointestinal dysfunction closely related with the menstrual cycle, such as distension, satiety, bloating and abdominal pain.
Animals ; Calcitonin Gene-Related Peptide ; blood ; Cholecystokinin ; blood ; Estradiol ; analogs & derivatives ; pharmacology ; physiology ; Female ; Gastric Emptying ; drug effects ; physiology ; Ovariectomy ; Progesterone ; pharmacology ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitonin Gene-Related Peptide ; metabolism ; Receptors, Cholecystokinin ; metabolism ; Stomach ; metabolism ; physiology

The effects of adrenomedullin (ADM) on intracellular calcium concentration ([Ca(2+)](i)) were investigated in cultured hippocampal neurons. Changes in [Ca(2+)](i) were detected by laser scanning confocal microscopy using Fluo 3-AM as the calcium fluorescent probe. [Ca(2+)](i) was represented by relative fluorescent intensity. The results showed that: (1) ADM (0.01-1.0 micromol/L) decreased the resting [Ca(2+)](i) in a concentration-dependent manner. (2) Calcitonin gene-related peptide receptor antagonist CGRP(8-37) significantly inhibited the effects of ADM. (3) ADM significantly reduced the increase in [Ca(2+)](i) induced by high K(+). (4) ADM markedly inhibited the inositol 1,4,5-trisphosphate (IP(3))-induced increase in [Ca(2+)](i), while did not influence ryanodine-evoked increase in [Ca(2+)](i). These results suggest that ADM reduces [Ca(2+)](i) in cultured hippocampal neurons through suppressing Ca(2+) release from IP(3)-sensitive stores. Although ADM does not alter resting Ca(2+) influx, it significantly suppresses Ca(2+) influx activated by high K(+). These effects may be partly mediated by CGRP receptors. ADM in the CNS may act as a cytoprotective factor in ischemic/hypoxic conditions.
Adrenomedullin ; Animals ; Animals, Newborn ; Calcitonin Gene-Related Peptide ; metabolism ; Calcium ; metabolism ; Cells, Cultured ; Embryo, Mammalian ; Hippocampus ; cytology ; metabolism ; Inositol 1,4,5-Trisphosphate ; antagonists & inhibitors ; Neurons ; cytology ; metabolism ; Peptides ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitonin Gene-Related Peptide ; antagonists & inhibitors ; metabolism

OBJECTIVETo investigate the value of CD64 mean fluorescence intensity of the peripheral blood neutrophils as a diagnostic marker of bacterial infection in patients with hematologic malignancies after chemotherapy.

METHODSThe neutrophil CD64 mean fluorescence intensity of all patients was detected by flow cytometry, and compared with procalcitonin (PCT) and C reactive protein (CRP) detected in part of patients; the relationship between nCD64 and bacterial infection were analyzed through continuous dynamic monitoring nCD64 mean fluorescence intensity in part of patients.

RESULTSThe expression of nCD64 was not affected by neutrophils counts (P>0.01); the nCD64 mean fluorescence intensity, PCT and CRP levels in infection group and dynamic monitoring group were significantly higher than those in non-infected group (P<0.01); the sensitivity and specificity of nCD64 mean fluorescence intensity were much higher, as compared with PCT and CRP in diagnosis of bacterial infection.

CONCLUSIONnCD64 mean fluorescence intensity can be used as an effective diagnostic marker for bacterial infection in patients with hematologic malignancies after chemotherapy, and may be used to forecast bacterial infection to a certain extent.

Antineoplastic Agents ; Bacterial Infections ; C-Reactive Protein ; Calcitonin ; Calcitonin Gene-Related Peptide ; Flow Cytometry ; Hematologic Neoplasms ; Humans ; Neutrophils ; Protein Precursors ; Receptors, IgG

OBJECTIVE: We studied the expression of pain-related molecules such as substance P involved in chronic pain of fibromyalgia syndrome (FMS) patients using rat cortical cells in hypoxia. METHODS: We sacrificed pregnant Sprague-Dawley rat and got embryo. We cultured the cortical cells and compared the expression of pain-related molecules in 1st, 3rd, 5th day cortical cells exposed to hypoxia (37 degrees C, 5% CO2, 98% N2) to control by immunohistochemistry. We measured the density at soma using softwear 'Scion image'. RESULTS: The expression of substance P was increased in hypoxic cortical cell group than control (control mean: 49.9 vs. hypoxia 1st day: 75.4 (p<0.001), 3rd day: 65.6 (p<0.001), 5th day: 79.9 (p<0.001)). The expression of kainate receptor was increased in hypoxic cortical cell group than control (control mean: 58.4 vs. hypoxia 1st day: 64.9 (p<0.001), 3rd day: 63.3 (p<0.001), 5th day: 62.9 (p<0.001)). The expression of N-methyl-D-aspartate receptor 2B was increased in hypoxic cortical cell group than control (control mean : 59.4 vs. hypoxia 1st day: 60.8 (p<0.001), 3rd day: 62.6 (p<0.001), 5th day: 67.1 (p<0.001)). But, the expression of calcitonin gene related peptide was decreased in hypoxic cortical cell group than control (control mean: 76.8 vs. hypoxia 1st day: 76.4 (p<0.001), 3rd day: 71.5 (p<0.001), 5th day: 61.3 (p<0.001)). CONCLUSION: Hypoxia during night could increase the expression of some pain-related molecules, which might be the cause of chronic pain in FMS patients.
Animals ; Anoxia ; Calcitonin Gene-Related Peptide ; Carisoprodol ; Chronic Pain ; Embryonic Structures ; Fibromyalgia* ; Glutamic Acid ; Humans ; Immunohistochemistry ; N-Methylaspartate ; Rats ; Rats, Sprague-Dawley ; Receptors, Kainic Acid ; Substance P