Cancer pain is one of the most common symptoms in patients with advanced cancer. In this study, we aimed to investigate the effects of the Mas-related gene C (MrgC) receptors on bone cancer pain. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured after the inoculation of Walker 256 mammary gland carcinoma cells into the tibia of adult Sprague-Dawley rats. The effects of MrgC receptor agonist bovine adrenal medulla 8-22 (BAM8-22) on nociceptive behaviors were investigated after intrathecal injection on days 16 and 17. Glial fibrillary acidic protein (GFAP)-positive cells in the spinal dorsal cord, and calcitonin gene related peptide (CGRP)-, neuronal nitric oxide synthase (nNOS)- and IL-1β-positive neurons in the dorsal root ganglia (DRG) were examined by immunofluorescence staining. The expression of nNOS and IL-1β proteins in the spinal dorsal horn and the DRG was examined by Western blotting after treatment with (Tyr6)-γ2-MSH-6-12 (MSH), which was another MrgC receptor agonist. The results showed that intrathecal injection of BAM8-22 (30 nmol) attenuated mechanical allodynia in a rat model of bone cancer pain and the effects could last for about 60 min, and single administration of BAM8-22 for two consecutive days reduced mechanical allodynia by about half on the third day. Moreover, the number of GFAP-positive cells in the spinal dorsal horn, and the number of CGRP-, nNOS- and IL-1β-positive neurons in the DRG were decreased. Similarly, intrathecal administration of MSH (15 nmol) reduced the expression of nNOS and IL-1β in the spinal dorsal horn and the DRG. In conclusion, activation of MrgC receptors suppresses the activation of astrocytes in the spinal dorsal cord and the expression of CGRP, nNOS, and IL-1β in the spinal dorsal cord and/or DRG, which may underlie the inhibition of bone cancer pain. These findings provide a novel strategy for the treatment of bone cancer pain.
Animals ; Cancer Pain/drug therapy* ; Rats ; Rats, Sprague-Dawley ; Bone Neoplasms/complications* ; Ganglia, Spinal/metabolism* ; Spinal Cord Dorsal Horn/metabolism* ; Receptors, G-Protein-Coupled/genetics* ; Female ; Calcitonin Gene-Related Peptide/genetics* ; Interleukin-1beta/metabolism* ; Peptide Fragments/metabolism* ; Nitric Oxide Synthase Type I/genetics* ; Disease Models, Animal

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

PURPOSE: The urinary bladder (UB) is innervated by both sensory and autonomic nerves. Recent studies have shown that sensory neuropeptides induced contractions in the detrusor muscle. Therefore, in a mouse model, we investigated the presence of interactions between the submucosal sensory nerves and the autonomic nerves that regulate the motor function of the detrusor muscle. METHODS: UB samples from male C57BL/6 mice were isolated, cut into strips, and mounted in an organ bath. Dose-response curves to norepinephrine and phenylephrine were studied in UB strips with and without mucosa, and the effects of preincubation with a receptor antagonist and various drugs on relaxation were also studied using tissue bath myography. RESULTS: Phenylephrine-induced relaxation of the UB strips showed concentration-related effects. This relaxation appeared in both mucosa-intact and mucosa-denuded UB strips, and was significantly inhibited by lidocaine, silodosin, and guanethidine (an adrenergic neuronal blocker). Meanwhile, phenylephrine-induced relaxation was inhibited by pretreatment with propranolol and calcitonin gene-related peptide (CGRP)–depletory capsaicin in UB strips with and without mucosa. CONCLUSIONS: The present study suggests that phenylephrine activates the α-1A adrenergic receptor (AR) of the sensory nerve, and then activates capsaicin-sensitive sensory nerves to release an unknown substance that facilitates the release of norepinephrine from adrenergic nerves. Subsequently, norepinephrine stimulates β-ARs in the detrusor muscle in mice, leading to neurogenic relaxation of the UB. Further animal and human studies are required to prove this concept and to validate its clinical usefulness.
Adrenergic Neurons ; Animals ; Autonomic Pathways ; Baths ; Calcitonin Gene-Related Peptide ; Capsaicin ; Guanethidine ; Humans ; Lidocaine ; Male ; Mice ; Mucous Membrane ; Myography ; Neuropeptides ; Norepinephrine ; Phenylephrine ; Propranolol ; Receptors, Adrenergic ; Receptors, Adrenergic, alpha-1 ; Relaxation ; Urinary Bladder*

BACKGROUND/AIMS: Previous studies have revealed that mast cells (MCs) may activate the protease-activated receptors and release of neuropeptides involved in the pathogenesis of irritable bowel syndrome (IBS). The levels of protease-activated receptor 2 (PAR-2) and tryptase can contribute to understanding the pathogenesis of IBS. METHODS: Colonoscopic biopsies were performed of 38 subjects (20 with IBS-diarrhea [IBS-D], eight with IBS-constipation [IBS-C], and 10 healthy volunteers). The mRNA and protein levels of tryptase and PAR-2 were assessed by real-time PCR and Western blot. The levels of vasoactive intestinal peptide (VIP), substance P (SP), and calcitonin gene-related peptide (CGRP) were measured by immunohistochemistry, and MCs were counted by toluidine blue staining. RESULTS: Significant increases in the mRNA expression of tryptase (p<0.05, IBS-D, IBS-C vs control) and PAR-2 (p<0.05, IBS-D, IBS-C vs control) and in the tryptase protein level (p<0.05, IBS-D, IBS-C vs control) were detected in IBS. Elevations of MCs, CGRP, VIP and SP (p<0.05, IBS-D vs control) were observed for IBS-D only. CONCLUSIONS: Tryptase levels may upregulate the function of PAR-2, resulting in the release of neuropeptide and they were correlated with clinical symptoms associated with IBS.
Biopsy ; Blotting, Western ; Calcitonin Gene-Related Peptide ; Immunohistochemistry ; Inflammation ; Irritable Bowel Syndrome* ; Mast Cells ; Neuropeptides ; Real-Time Polymerase Chain Reaction ; Receptor, PAR-2* ; Receptors, Proteinase-Activated ; RNA, Messenger ; Substance P ; Tolonium Chloride ; Tryptases* ; Vasoactive Intestinal Peptide

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

The increase of pronociceptive mediators in the dorsal root ganglia (DRG) and spinal dorsal horn is an important mechanism in the pathogenesis of inflammatory pain and opioid tolerance. Adrenomedullin (AM) belongs to calcitonin gene-related peptide (CGRP) family and has been recently demonstrated to be a pain-related peptide. It has also been shown that the expression and release of AM are increased in the DRG and spinal dorsal horn during inflammation and repeated use of morphine. Intrathecal administration of the selective AM receptor antagonist AM22-52 abolishes inflammatory pain and morphine tolerance, suggesting that enhanced AM receptor signaling in the DRG and spinal dorsal horn contributes to the induction of inflammatory pain and morphine tolerance. The present review highlights the recent developments regarding the involvement of AM in these two disorders. The neurological mechanisms of AM's actions are also discussed.
Adrenomedullin ; pharmacology ; Animals ; Calcitonin Gene-Related Peptide ; Drug Tolerance ; Ganglia, Spinal ; drug effects ; Inflammation ; drug therapy ; metabolism ; Morphine ; pharmacology ; Pain ; drug therapy ; metabolism ; Peptide Fragments ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenomedullin ; metabolism

OBJECTIVE: To investigate the effect of intrathecal sufentanil and protein kinase C inhibitor on pain threshold and the expression of N-methyl-D-aspartate receaptors (NMDAR)/calcitonin generelated peptide (CGRP) in spinal dorsal horn in rats with neuropathic pain. METHODS: Fifty-four healthy male Sprague-Dawley rats were randomly divided into 6 groups (9 in each group). The rats in the sham group(Group S) + spared nerve injury (SNI), SP+SNI, and P+SNI were intrathecally injected sufentanil (1 μg), sufentanil (1 μg) and chelerythrine chloride (11 μg), chelerythrine chloride (11 μg) followed by 10 μL normal saline once every day for 14 days postoperatively, respectively. Similarly, rats in the control group (Group C), the sham group (Group S), and SNI model group (Group SNI) were intrathecally injected 20 μL normal saline in the uniform interval. Pain behaviours were measured on Day 1 pre-surgery and on Day 1, 2, 7, and 14 after the intrathecal injection. The expressions of NMDAR and CGRP in the spinal dorsal horn of L5 segment were determined by immunohistochemistry on Day 2, 7, and 14 after the intrathecal injection. RESULTS: Compared with Group C and Group S, mechanical allodynia threshold in group SNI was decreased after the surgery (P<0.01), and expressions of NMDAR and CGRP immunoreactive soma in the spinal dorsal horn was significantly increased (P<0.01). Mechanical stimulation pain threshold was elevated in Group S+SNI, Group P+SNI, and Group SP+SNI compared with Group SNI (P<0.01), while expressions of NMDAR and CGRP immunoreactive soma in Group S+SNI, Group P +SNI, and Group SP+SNI were significantly decreased (P<0.05 or 0.01). CONCLUSION Intrathecal administration of sulfentanil and protein kinase C inhibitor can provide significant antinociception in rats with neuropathic pain and obviously inhibit the upregulation of NMDAR and CGRP expressions in the spinal dorsal horn of SNI rat models.
Animals ; Benzophenanthridines ; administration & dosage ; Calcitonin Gene-Related Peptide ; metabolism ; Injections, Spinal ; Male ; Neuralgia ; drug therapy ; metabolism ; physiopathology ; Pain Measurement ; Posterior Horn Cells ; metabolism ; Protein Kinase C ; antagonists & inhibitors ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Sufentanil ; administration & dosage

The present study investigated the effects of intrathecal (i.t.) application of bovine adrenal medulla 22 (BAM22), an endogenous opioid peptide potently activating opioid receptors and sensory neuron-specific receptor (SNSR), on a model of complete Freund's adjuvant (CFA)-induced inflammatory pain. Unilateral, but not bilateral, inflammatory pain was induced by intraplantar (i.pl.) injection of CFA in one side, as indicated by the shortened paw withdrawal latency and the increased edema of paw. Paw withdrawal latency test, paw edema determination and immunohistochemistry were used in CFA-induced inflammatory pain model after i.t. administration of BAM22 or saline. It was found that administration of BAM22 dose-dependently attenuated CFA-induced hyperalgesia and edema, and resumed antinociceptive effects against thermal stimulation in behavioral test. In 10 nmol BAM22 group, paw withdrawal latency was resumed to 83.2% of normal, and edema increased only by 60% of normal at 48 h. The potency of BAM22 was 33.5% of maximal possible effect (MPE) at 24 h, and the antinociception persisted for at least 1 h. Furthermore, i.t. treatment of 10 nmol BAM22 evidently decreased the expressions of CFA-evoked neuronal nitric oxide synthase (nNOS)-positive cells and calcitonin gene-related peptide (CGRP)-immunoreactivity positive nerve fibers by 25.6% (P<0.01) and 25.2% (P<0.001) compared with saline group, respectively, at L3-L5 segments of the spinal cord. Small and medium CGRP-positive cells were 57.4% and 35.2% in dorsal root ganglion (DRG) in 10 nmol BAM22 group, respectively, which were remarkably lower than those in saline group (P<0.001). The present study suggests that BAM22 relieves CFA-induced thermal hyperalgesia in the early phase and resumes antinociceptive effects through down-regulation of nNOS and CGRP expressions in DRG and spinal cord, which is possibly mediated via SNSR.
Animals ; Calcitonin Gene-Related Peptide ; metabolism ; Enkephalin, Methionine ; analogs & derivatives ; pharmacology ; Freund's Adjuvant ; Hyperalgesia ; etiology ; physiopathology ; Inflammation ; chemically induced ; complications ; Male ; Nitric Oxide Synthase Type I ; metabolism ; Pain ; etiology ; physiopathology ; Pain Measurement ; drug effects ; Protein Precursors ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; physiology

BACKGROUND: The pathogenesis of melasma has not yet been clearly demonstrated. But, clinical manifestations such as remarkable lesional symmetry and the distribution related to trigeminal nerves, suggest that the neural system could play a pathogenic role in melasma. OBJECTIVE: This study was carried out to examine the expression of some neuropeptides and their receptors, which are well known to be major contributors of neuroinflammation in many dermatoses, in melasma lesions. METHODS: Skin biopsies were obtained from the lesional and non-lesional facial skin of 6 Korean women with melasma. Immunofluorecence staining and confocal laser scanning microscopy were performed. RESULTS: In our results, no difference could be detected with regard to the intensity of immunoreactivity for vasoactive intestinal peptide (VIP), calcitonin gene-related peptide (CGRP), calcitonin gene-related peptide receptor (CGRPR), substance P (SP), substance P receptor (SPR), somatostatin (SOM), pituitary adenylate cyclase activating peptide (PACAP) and pituitary adenylate cyclase activating peptide receptor (PACAPR) in the lesional skins compared with the non-lesional skins. CONCLUSION: These results suggest that neuroinflammation induced by neuropeptides such as substance P, calcitonin gene-related peptide, vasoactive intestinal peptide, and somatostatin and their receptors included in this study, are not directly associated with melasma pathogenesis.
Adenylyl Cyclases ; Biopsy ; Calcitonin Gene-Related Peptide ; Female ; Humans ; Melanosis ; Microscopy, Confocal ; Neuropeptides ; Receptors, Neurokinin-1 ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide ; Skin ; Skin Diseases ; Somatostatin ; Substance P ; Trigeminal Nerve ; Vasoactive Intestinal Peptide

In this study, the expressions of growth hormone secretagogue receptor type 1a (GHS-R1a) in the rat dorsal root ganglion (DRG) and nodose ganglion (NG) were investigated by using immunohistochemistry and in situ hybridization. The results clearly showed the presence of GHS-R1a mRNA and GHS-R1a-positive neurons in the rat DRG and NG. GHS-R1a was also co-localized with calcitonin gene-related peptide (CGRP) in some DRG and NG neurons, indicating the existence of subpopulations of the visceral afferents. The extrinsic primary afferent visceroceptive DRG and NG neurons from the stomach were identified by retrograde tracing fluorogold and stained for GHS-R1a and CGRP. Some neurons both positive for CGRP and GHS-Rla were labled by fluorogold. Our results not only demonstrate the expression of GHS-R1a in the vagal afferents but also provide the first and direct morphological evidence for its presence in the spinal visceral afferents, and gherin might have a modulatory role in the visceral afferent signaling.
Afferent Pathways ; Animals ; Calcitonin Gene-Related Peptide ; metabolism ; Ganglia, Spinal ; cytology ; Immunohistochemistry ; Neurons, Afferent ; cytology ; Nodose Ganglion ; cytology ; Rats ; Receptors, Ghrelin ; metabolism ; Stomach ; innervation