This article describes the preparation of salmon calcitonin ultra-flexible liposomes and their hypocalcemia effect after intranasal administration in rats. Both the conventional liposomes and ultra-flexible liposomes were prepared by rotary evaporation-sonication and extrusion. The morphology of ultra-flexible liposomes was observed with transmission electronic microscope. The size and size distribution and their zeta potential were determined by dynamic light scattering. The mean size of ultra-flexible liposomes with DC-Chol was no more than 120 nm, while the mean size of the conventional liposomes was 256.5 nm. The results showed the content of sodium deoxycholate have significant effect on the mean particle size of liposomes. The ultra-flexible liposomes were intranasal administrated at the dose of 5.0 microg x kg(-1); the concentration of serum calcium was determined by OCPC method. The results showed that the salmon calcitonin solution only slightly lowered serum calcium levels and the conventional liposomes could improve the effect of decreased serum calcium level (D%), and the ultra-flexible liposomes had the best effect on the decreased serum calcium level, and the hypocalcemia effect was correlated with the content of sodium deoxycholate which was present in the liposomes. Moreover the ciliotoxicity of ultra-flexible nanoliposomes on nasal mucocilia was investigated with the electron microscope scanning. The results showed that the ultra-flexible liposomes markedly reduced the ciliotoxicity of sodium deoxycholate on nasal mucous. Thereby the ultra-flexible liposomes significantly enhanced the hypocalcemia effect of serum calcium after intranasal administration in rats. The ultra-flexible liposomes could be an effective carrier for intranasal delivery of the peptide and protein drugs.
Administration, Intranasal ; Animals ; Calcitonin ; administration & dosage ; pharmacology ; Calcium ; blood ; Liposomes ; Male ; Particle Size ; Rats ; Rats, Sprague-Dawley

AIMTo prepare salcatonin dry powder inhalations (sCT-DPIs) A (mixture of mannitol and L-leucine) and B (mixture of manntiol and lactose) by spray-drying and then to study their main pharmaceutical properties.

METHODSDumping rate of sCT-DPIs capsules and deposited fraction of sCT at effective part were determined according to Chinese Pharmacopiea 2000. Particle morphology under different relative humidity (RH) was observed by scanning electronics microphotograph, particle size and its distribution were determined by Malvern Mastersizer and the transition of morphorous state for carriers before and after spray-drying was investigated by differential thermal analysis (DTA) and X-ray powder diffraction (XRPD).

RESULTSDumping rates of sCT-DPIs A and B capsules were both above 10% and deposited fraction of sCT at effective part was above 90% for both A and B, which were all in agreement with the standard of Chinese Pharmacopiea 2000. Powder particle of sCT-DPIs A was round and existed one by one after keeping one month under RH 0, 23% and 52%, but aggregation can be observed under RH 75%; many particles which were also round agglomerated in sCT-DPIs B even under zero RH; mean particle size of sCT-DPIs A was 1.67 microns, which was much smaller than that of sCT-DPIs B; In sCT-DPIs A particle with empty core which was lighter than the same size particle with concreted core was observed. It was shown by DTA that melted heat of L-leucine in sCT-DPIs composed of mannitol and L-leucine lowered much more than that of L-leucine exisited alone after spray-drying. It was confirmed by XRPD that diffraction intensity of carriers in sCT-DPIs decreased more than that of carriers before spray-drying.

CONCLUSIONRound particle can be made when mannitol was added to carriers and ultra low density carriers can be formed when L-leucine was added. It was suggested by SEM that DPIs should be kept under certain RH. Particle size and distribution of sCT-DPIs all accorded with demand of DPIs. Complex spray-drying carriers formed amorphous state easier than single carrier.

Administration, Inhalation ; Anti-Asthmatic Agents ; administration & dosage ; Calcitonin ; administration & dosage ; Differential Thermal Analysis ; Leucine ; Mannitol ; Microscopy, Electron, Scanning ; Particle Size ; Powders ; Technology, Pharmaceutical ; methods

OBJECTIVETo explore the effects of oral administration of mixed enteral nutritional agent on intestinal mucosal barrier of patients with severe burn injury at early stage.

METHODSTwenty-four patients with severe burn injury admitted to our burn ward from August 2013 to September 2014, conforming to the study criteria, were divided into conventional therapy group (n = 12) and early enteral feeding group (n = 12) according to the random number table. Patients in conventional therapy group received conventional treatment immediately after admission, while those in early enteral feeding group were orally given 100 mL of a mixture of glutamine, probiotics, and prebiotics once a day besides conventional treatment for 7 days. Serum levels of diamine oxidase (DAO) and procalcitonin (PCT) and plasma level of LPS were determined by ELISA before treatment and on treatment day (TD) 1, 3, 7, 14, and 21. Wound secretion and blood samples were collected for bacterial culture within the 21 TD. The incidence of MODS within the 21 TD was observed. Data were processed with Fisher's exact test, rank sum test, analysis of variance for repeated measurement, and LSD-t test.

RESULTS(1) Serum levels of DAO in patients of early enteral feeding group on TD 7, 14, and 21 were respectively (14.9 ± 3.7), (12.4 ± 3.1), and (9.5 ± 0.7) ng/mL, which were significantly lower than those of conventional therapy group [(17.5 ± 4.0), (16.3 ± 3.3), and (13.0 ± 1.1) ng/mL, with t values from 2.913 to 15.304, P values below 0.01]. Serum levels of DAO at the other time points were close between the two groups (with t values from -0.598 to 0.139, P values above 0.05). (2) Compared with serum levels of PCT in patients of conventional therapy group [(11.7 ± 20.9) and (12.9 ± 23.9) ng/mL], those of early enteral feeding group were significantly lower on TD 7 and 14 [(2.7 ± 8.1) and (2.0 ± 5.6) ng/mL, with Z values respectively -2.919 and -2.139, P < 0.05 or P < 0.01]. Serum levels of PCT at the other time points were close between the two groups (with Z values from -1.833 to -0.346, P values above 0.05). (3) Plasma level of LPS in patients of early enteral feeding group on TD 7 was (33 ± 56) pg/mL, which was significantly lower than that of conventional therapy group [(102 ± 108) pg/mL, Z = -2.046, P < 0.05]. Plasma levels of LPS at the other time points between the two groups showed no significant difference (with Z values from -2.003~-0.526, P values above 0.05). (4) Positive results in bacterial culture of wound secretion were approximately the same between the two groups (P > 0.05). Bacterial culture of blood was positive in 7 patients of conventional therapy group and 1 patient of early enteral feeding group, showing significantly statistical difference (P < 0.05). MODS was observed in 1 patient of conventional therapy group, showing no significantly statistical difference with that of early enteral feeding group (no patient, P > 0.05).

CONCLUSIONSEarly intestinal feeding of mixed enteral nutritional agent in addition to conventional therapy can effectively promote repair of the impairment of intestinal mucosal barrier, protect integrity of intestinal mucosa, reduce damage to intestines, and alleviate inflammatory response in patients suffering from severe burn injury.

Administration, Oral ; Amine Oxidase (Copper-Containing) ; blood ; Burns ; metabolism ; therapy ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Enteral Nutrition ; methods ; Female ; Glutamine ; administration & dosage ; pharmacology ; Humans ; Intestinal Mucosa ; drug effects ; metabolism ; Protein Precursors ; blood ; Treatment Outcome ; Wound Healing

BACKGROUNDHigh fluoride exposure can result in dental fluorosis. Fluoride and iodine are coexistent in the drinking water of areas in China and may affect the prevalence of dental fluorosis and osteogenesis. The aim of this study was to investigate the relationship between serum calciotropic hormone level, and dental fluorisis in children exposed to different concentrations of fluoride and iodine in drinking water.

METHODSA pilot study was conducted in three villages located in the Kaifeng and Tongxu counties of Henan Province, China in 2006. Children aged 8 to 12 years, born and raised in the three villages were recruited. The fluoride levels in the samples of urine from these children were detected by fluoride ion selective electrode. Calcitonin and osteocalcin levels in the serum, and serum calcium were measured by radioimmunassay and flame atomic absorption spectrometry, respectively.

RESULTSFluoride levels in urine were significantly lower in children from control area (CA) as compared with those from the high fluoride & iodine areas (HFIA) and the high fluoride area (HFA) (P < 0.05 respectively), and no statistically significant difference was found between the children from HFIA and HFA. Additionally, calcitonin levels in the serum were significantly lower in children from CA and HFA as compared with that from HFIA (P < 0.05 respectively), and osteocalcin levels in the serum was lower in children from CA than those from HFIA (P < 0.05). No statistically significant difference in serum osteocalcin concentrations was found between children from HFA and HFIA.

CONCLUSIONThis study provides an evidence that iodine exposure may modify the serum calciotropic hormone levels related to fluorine exposure.

Calcitonin ; blood ; Child ; Female ; Fluorides ; administration & dosage ; Fluorosis, Dental ; epidemiology ; Humans ; Iodine ; administration & dosage ; Male ; Osteocalcin ; blood ; Pilot Projects ; Prevalence ; Water Supply ; analysis

Visceral afferent nerve fibers containing substance P or calcitonin gene-related peptide (CGRP) are distributed in the bladder wall, and are known to be stimulated by and then desensitized by capsaicin. Recently, there have been some reports on the effectiveness of intravesical capsaicin administration for the treatment of hypersensitive lower urinary tract disorder or neurogenic bladder. In this study, the effects of intravesical capsaicin on the substance P or CGRP immunoreactivities in the spinal dorsal horn were investigated and the mechanism of capsaicin treatment for bladder disorders was revealed. After intravesical administration of capsaicin, the substance P and CGRP immunoreactive areas were measured at the dorsal horn of L4 and S1 spinal cord. Before capsaicin treatment, the substance P immuno- reactive area was 2.61+/-0.78 x 105 mm2 in L4 and 1.66+/-0.49 x 105 mm2 in S1. The substance P immunoreactivity was markedly reduced 1~2 weeks after capsaicin treatment in both L4 and S1 spinal cord. The CGRP immunoreactive area was 1.74+/-0.52 x 105 mm2 in L4 and 1.14+/-0.69 x 105 mm2 in S1, but was not reduced after capsaicin treatment. Therefore, capsaicin administered intravesically desensitizes nerve fibers containing substance P and consequently suppresses pain and voiding reflex.
Administration, Intravesical ; Animals ; Calcitonin Gene-Related Peptide ; Capsaicin* ; Horns* ; Nerve Fibers ; Rats* ; Reflex ; Spinal Cord ; Substance P ; Urinary Bladder ; Urinary Bladder, Neurogenic ; Urinary Tract ; Visceral Afferents

OBJECTIVETo discuss the effect of calcitonin gene-related peptides (CGRP) on epithelial cadherin (E-cd) expression in human bronchial epithelial cells (HBECs) in vitro.

METHODSThe effect of CGRP on E-cd protein and mRNA expression in both normal and O3-challenged HBECs were determined by immunocytochemistry and RT-PCR. The signal transduction pathways of CGRP were observed by using protein kinase C(PKC) inhibitor (H-7), calmodulin(CaM) inhibitor (W-7) and PKA inhibitor (H-89).

RESULTSCGRP increased E-cd mRNA and protein expressions of normal and O3-challenged HBECs in a dose-dependent manner. CGRP had no effect on cytoplasm E-cd expression. Pre-treatment with H-89, H-7 and W-7, the up-regulatory effect of CGRP on E-cd expression was partly abolished.

CONCLUSIONCGRP increased in cytomembrane E-cd expression of normal and O3-challenged HBECs in a dose-dependent manner. E-cd expression on HBECs was strengthened by CGRP via PKA, PKC and CaM pathways.

Bronchi ; cytology ; Cadherins ; metabolism ; Calcitonin Gene-Related Peptide ; administration & dosage ; pharmacology ; Cell Line ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Ozone ; RNA, Messenger ; genetics

Spontaneous pain, allodynia and hyperalgesia are well known phenomena following peripheral nerve or tissue injury, and it is speculated that secondary hyperalgesia and allodynia, are generally thought to depend on a hyperexcitability (sensitization) of neurons in the dorsal horn. It is supposed that the sensitization may be due to various actions of neurotransmitters (SP, CGRP, excitatory amino acids) released from the primary afferent fibers. In this study, we examined effects of the iontophoretically applied SP and CGRP on the response to EAA receptor agonists (NMDA and non-NMDA) in the WDR dorsal horn neurones and see if the effects of SP or CGRP mimic the characteristic response pattern known in various pain models. The main results are summarized as follows: 1) SP specifically potentiated NMDA response. 2) CGRP non-specifically potentiated both NMDA and AMPA responses. Potentiation of NMDA response, however, was significantly greater than that of AMPA response. 3) 50% of SP applied cells and 15.8% of CGRP applied cells showed reciprocal changes(potentiation of NMDA response and suppression of AMPA response). These results are generally consistent with the sensitization characteristics in diverse pain models and suggests that the modulatory effects of SP and CGRP on NMDA and non-NMDA (AMPA) response are, at least in part, contribute to the development of sensitization in various pain models.
Animal ; Calcitonin Gene-Related Peptide/pharmacology* ; Calcitonin Gene-Related Peptide/administration & dosage ; Excitatory Amino Acid Agonists/pharmacology* ; Iontophoresis ; Male ; N-Methylaspartate/pharmacology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord/physiology ; Spinal Cord/drug effects* ; Substance P/pharmacology* ; Substance P/administration & dosage ; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid/pharmacology

BACKGROUNDProstatodynia remains a difficult clinical problem. Resiniferatoxin (RTX), an ultrapotent vanilloid, can produce a selective and long-lasting desensitization of nociception via C-fiber sensory neurons. Substance P (SP) and calcitonin gene-related peptide (CGRP) released from C-fibers are key neurotransmitters in visceral pain. In this study, we evaluated the analgesic effect of intrathecal RTX on rat prostatodynia.

METHODSMale Sprague-Dawley rats were divided into 3 groups for different treatment. In group A, sham operation was preformed. In group B, 100 microl complete Freund's adjuvant (CFA) was injected into the rat's bilateral ventral prostate to induce chronic inflammation. In group C, after prostatitis formed, 50 microl 10 nmol/L RTX was injected into the rat's lumbosacral (L5-S2) vertebral canal. SP and CGRP contents in the spinal cord were investigated by immunohistochemistry and radioimmunoassay (RIA). Their transcriptional levels in dorsal root ganglion (DRG) were determined by reverse transcriptase polymerase chain reaction (RT-PCR). In addition, pelvic nerve afferent discharge was recorded to explore the neuro-electrophysiological mechanisms underlying RTX-induced effect.

RESULTSSP and CGRP released in the spinal cord and their synthesis in DRG were increased significantly in response to CFA-induced chronic prostatitis, whereas this increase was effectively inhibited by intrathecal RTX. Meanwhile, pelvic nerve afferent electrical activity was enhanced significantly in rats with chronic prostatitis, but it was attenuated markedly in RTX-treated rats paralleled by the change of neuropeptides.

CONCLUSIONSIntrathecal RTX administration could produce an analgesic effect on rat prostatodynia. Suppression of pelvic nerve afferent electrical activity may be a crucial mechanism underlying RTX-induced analgesia. RTX intrathecal application may present a novel analgesic strategy of prostatodynia.

Analgesics ; administration & dosage ; Animals ; Calcitonin Gene-Related Peptide ; analysis ; genetics ; Diterpenes ; administration & dosage ; Injections, Spinal ; Male ; Prostatitis ; drug therapy ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Substance P ; analysis ; genetics

BACKGROUNDRenal osteodystrophy is one of the commonest complications of chronic renal failure. It may have a severe impact on the quality of life of patients on maintenance dialysis therapy. Besides post-menopausal women and elderly people, the dialysis patients are another high risk group. But at present, there is no research on how to prevent osteoporosis in maintenance dialysis patients. This study was conducted to observe the bone density of maintenance dialysis patients and to evaluate the clinical outcomes and safety of different administration dosage of salmon calcitonin to prevent osteoporosis in maintenance dialysis patients.

METHODSOne hundred and forty-eight patients on maintenance dialysis were involved in the 12-month, randomized, controlled trial. Fifty patients (experiment I group) received subcutaneous injection of salmon calcitonin (50 U) three times a week for 12 months. Fifty patients (experiment II group) received subcutaneous injection of salmon calcitonin (100 U) three times a week for 12 months. At the same time, both of them received oral calcium carbonate 1500 mg tid and rocaltrol 0.25 microg qn for 12 months. The control group only received oral calcium carbonate 1500 mg tid and rocaltrol 0.25 microg qn for 12 months. The levels of bone mass density (BMD) of the lumbar spine and femoral neck, serum intact parathyroid hormone (iPTH), osteocalcin (OC), calcium, phosphorus, alkaline phosphatase (ALP) were assessed at baseline and then again after 3, 6 and 12 months of treatment.

RESULTSThe values of BMD at the lumbar spine and femoral neck before the treatment were not significantly different from those 3, 6, and 12 months after the treatment in trial groups I and II (all P > 0.05) and there were no significant differences in the BMD values at different time points between trial groups I and II. In the control group, the BMD values at the lumbar spine and femoral neck 3, 6, and 12 months after the beginning of trial were significantly lower than those before the trial, and significantly lower than the corresponding values of trial groups I and II (all P < 0.05). The serum OC 3, 6, and 12 months after the treatment was significantly lower than that before the experiment (all P < 0.05) in the control group. However, there was no significant difference in the value of serum OC before and 3, 6, and 12 months after the treatment in trial groups I and II (all P > 0.05).

CONCLUSIONSThe dose of salmon calcitonin 50 U three times a week plus calcium carbonate and active vitamin D can effectively preserve the BMD and prevent bone loss in maintenance dialysis patients, and it is well tolerated by patients on maintenance dialysis.

Adult ; Alkaline Phosphatase ; blood ; Bone Density ; drug effects ; Bone Density Conservation Agents ; administration & dosage ; therapeutic use ; Calcitonin ; administration & dosage ; therapeutic use ; Calcium ; blood ; Calcium Carbonate ; administration & dosage ; therapeutic use ; Drug Administration Schedule ; Female ; Femur Neck ; drug effects ; metabolism ; Humans ; Lumbar Vertebrae ; drug effects ; metabolism ; Male ; Middle Aged ; Osteocalcin ; blood ; Osteoporosis, Postmenopausal ; blood ; prevention & control ; Parathyroid Hormone ; blood ; Phosphorus ; blood ; Renal Dialysis

The purpose of this study was to investigate the treatment efficacies of salmon calcitonin (SC) and estrogen in a type-I osteoporotic rat model. Sixty, 3-month-old, female Wistar rats were divided into six groups. The first group was used as the control, and the second a sham, the other four were surgically ovariectomized. 24 hours after the ovariectomy, they were either left untreated (OVX), or treated with an injection of either 17-beta estradiol (E2) 30 mcg/kg/24 hours, low-dose calcitonin (LDC) 10 IU/ kg/48 hours or high-dose calcitonin (HDC) 20 IU/kg/48 hours. 6 weeks later, the bone densities were measured by DEXA, the animals sacrificed and the femurs harvested for histomorphometric evaluation. The bone mineral densities (BMD) of the spine and proximal femur were lower in the OVX group, but only the values of the spine BMD were statistically significant. The BMD of the spine seemed to be preserved with all the treatments. The histomorphometric evaluation revealed that after the OVX the decrease in the trabecular volume was prevented by all the treatments. However, significant changes in the indices of bone formation were not shown. In conclusion, all the treatments prevented bone lost in the ovariectomized rats. Histopathological measurements of bone formation are unlikely to provide any evidence for the effects of these agents on the osteoblastic function. In the animal model of estrogen depletion, our results suggest that the calcitonin provides an important alternative therapy for postmenopausal osteoporosis.
Animals ; Bone Density/drug effects ; Calcitonin/*administration & dosage ; Comparative Study ; Dose-Response Relationship, Drug ; Estradiol/*therapeutic use ; Female ; Osteoporosis/*drug therapy/pathology/physiopathology ; Ovariectomy ; Rats ; Rats, Wistar ; Salmon ; Spine/physiopathology