Calcitonin Research Articles

Experimental Infusion of Calcitonin Gene–Related Peptide Can Induce Cluster Headache

Calcitonin gene–related peptide (CGRP) is involved in triggering migraine headaches and has been found in patients who are having cluster headaches. To

Role of TRPV1/CGRP signaling pathway in lidocaine postconditioning-induced reduction of myocardial ischemia-reperfusion injury in rats

Objective To evaluate the role of transient receptor potential vanillic acid subtype 1(TRPV1)/calcitonin gene-related peptide(CGRP) signaling pathway in lidocaine postconditioning-induced reduction of myocardial ischemia-reperfusion(I/R) injury in rats. Methods Forty healthy male Sprague-Dawley rats, aged 3 months, weighing 250-300 g, were divided into 5 groups(n=8 each) using a random number table method: sham operation group(group Sham), group I/R, lidocaine postconditioning group(group LP), lidocaine postconditioning plus CGRP8-37 group(group LP+ CGRP8-37), and lidocaine postconditioning plus capsazepine group(group LP+ Capz). Myocardial ischemia was induced by ligating the anterior descending branch of left coronary artery for 30 min, followed by 120-min reperfusion in anesthetized rats. Lidocaine 2 mg/kg was injected via the tail vein at 5 min before reperfusion in group LP. In group LP + CGRP8-37, lidocaine was intravenously injected, and CGRP receptor selective antagonist CGRP8-37 2 mg/kg was intravenously injected at the same time. In group LP+ Capz, lidocaine was injected intravenously, and TRPV1 blocker capsazepine 3 mg/kg was injected intravenously at the same time. A catheter was retrogradely implanted to the left ventricle, and heart rate(HR), left ventricular systolic pressure(LVSP), left ventricular end-diastolic pressure(LVEDP), and the maximum rate of increase or decrease in left ventricular pressure(±dp/dtmax) were continuously monitored and recorded. Blood samples were collected from the carotid artery at 120 min of reperfusion for determination of cardiac troponin I(cTnI), myoglobin(Myo) and creatine kinase-MB(CK-MB) concentrations in serum. Rats were then sacrificed for determination of myocardial infarct size. Results Compared with Sham group, the serum concentrations of cTnI, Myo and CK-MB were significantly increased, LVSP, + dP/dtmax and HR were decreased, and LVEDP and -dP/dtmax were increased in I/R group and LP group(P 0.05). Compared with group LP, the serum concentrations of cTnI and Myo, myocardial infarct size and LVEDP were significantly increased, and + dP/dtmax was decreased in group LP+ CGRP8-37, and the serum concentrations of cTnI and Myo and myocardial infarct size were significantly increased, LVSP and + dP/dtmax were decreased, and LVEDP was increased in group I/R+ Lido+ Capz(P<0.05). Conclusion The mechanism by which lidocaine postconditioning mitigates myocardial I/R injury is related to activating TRPV1/CGRP signaling pathway in rats. Key words: Lidocaine; Ischemia postconditioning; Myocardial reperfusion injury; Calcitonin gene-related peptide; TRPV cation channels

Endogenous calcitonin regulates lipid and glucose metabolism in diet-induced obesity mice

Calcitonin (CT) plays an important role in calcium homeostasis, and its precursor, proCT, is positively associated with the body mass index in the general human population. However, the physiological role of endogenous CT in the regulation of metabolism remains unclear. Knockout mice with gene-targeted deletion of exon 4 of Calca (CT KO) were generated by targeted modification in embryonic stem cells. Male mice were used in all experiments and were fed a slightly higher fat diet than the standard diet. The CT KO mice did not exhibit any abnormal findings in appearance, but exhibited weight loss from 15 months old, i.e., significantly decreased liver, adipose tissue, and kidney weights, compared with wild-type control mice. Furthermore, CT KO mice exhibited significantly decreased fat contents in the liver, lipid droplets in adipose tissues, serum glucose, and lipid levels, and significantly increased insulin sensitivity and serum adiponectin levels. CT significantly promoted 3T3-L1 adipocyte differentiation and suppressed adiponectin release. These results suggested that CT gene deletion prevents obesity, hyperglycemia, and hyperlipidemia in aged male mice. This is the first definitive evidence that CT may contribute to glucose and lipid metabolism in aged male mice, possibly via decreased adiponectin secretion from adipocytes.

Calcitonin as an anticalcification treatment for implantable biological tissues

Background and aim of the studyCalcification remains the major role of failure of implantable biomedical material and in particular of bioprosthetic valves. Various treatments have been proposed to mitigate calcification of glutaraldehyde-fixed bioprosthetic valves but none have succeeded in inhibiting or mitigating efficiently the calcification process of the implantable biological tissues. Since the discovery of calcitonin (CT) and its therapeutic role in treating hypercalcemic patients, CT has never been tried as an anticalcification treatment for biomaterials. It is postulated, that tissue calcification may be efficiently minimized by forming adducts with aldehyde groups thus eliminating the places of the biological tissues onto the calcium cations could be deposited. Material and methodsFresh porcine aortic leaflets were cut radially in three parts. Three groups of tissue were created. Group I (glutaraldehyde only), Group II (glutaraldehyde with 1% CT) and Group III (glutaraldehyde with 10% CT). All tissues were then implanted subdermally in three sets of 8 (Group I) and 9 (Group II and Group III) male Wistar rats of 12 days old. 21 days later the rats were euthanized by inhalation of CO2. The tissues were retrieved and after rinsing with distilled water 3 times, were lyophilized at −40°C at high vacuum pressure of approximately 100mmHg for 16h. The calcium content was then measured with flat atomic absorption technique. ResultsThe preimplantation values of Ca concentration as expressed in mg Ca/g of tissue were 1.79±0.14 in Group I, 4.78±0.0079 in Group II and 2.88±0.17 in Group III (p=ns). 21 days later the values of Ca concentration were 126.95±12.97 for Group I, 24.69±2.71 for Group II (p<0.05) and 27.16±2.95 for Group III (p<0.05). There was not significance difference between Groups II and III, even if Group II showed a less accumulation of Ca concentration (×5.16) than Group III (×9.43). ConclusionAn anticalcification treatment based on calcitonin as an additive to buffered glutaraldehyde, mitigates the calcification process of the implantable biological tissues, as compared to glutaraldehyde treatment only.

Detection and significance of calcitonin gene-related peptide in children with enterovirus 71 infected hand foot and mouth disease

Objective To detect the plasma calcitonin gene-related peptide (CGRP) in children with enterovirus 71(EV71) infected hand foot and mouth disease(HFMD), and explore the relationship between CGRP and the severity of EV71 infected HFMD. Methods Two hundred children with EV71 infected HFMD in Xi′an Children′s Hospital from January 2017 to December 2017 were selected as the research group, and 50 healthy children were selected as control group in the same period.According to the severity of the disease, the research group was divided into the mild group (n=142) and the severe group (n=58). The level of plasma CGRP was detected by ELISA. Results In acute stage, the level of CGRP in children with EV71 infected HFMD was lower than that in control group, and the difference was statistically significant (t=4.235, P<0.001). The level of CGRP in acute stage in severe group[(29.90±5.10)pg/ml] was significantly lower than that of the mild group[(35.51±5.48)pg/ml], and the difference was statistically significant (t=6.615, P=0.001). The level of CGRP in mild group[(35.51±5.48)pg/ml] was slightly lower than that of the control group[(36.63±5.65)pg/ml], but the difference was insignificant (t=1.120, P=0.086). In recovery stage, the level of CGRP in the severe group was obviously higher than that in the acute stage(t=5.924, P=0.003). According to the ROC curve of CGRP in mild and severe EV71 infected HFMD, the critical value of CGRP in mild and severe HFMD was 28.12 pg/ml, and the sensitivity was 80.0% and the specificity was 50.0%. Conclusion The CGRP is associated with the progression of EV71 HFMD, and when CGRP is lower than 28.12 pg/ml, suggesting the possibility of severe HFMD.The decrease of plasma CGRP may be a risk factor for the progression of EV71 infected HFMD. Key words: Hand foot and mouth disease; Enterovirus 71; Calcitonin gene-related peptide

Changes in expression of IB4 and CGRP in neurons in dorsal root ganglia of rats with neuropathic pain

Objective To evaluate the changes in the expression of isolectin B4 (IB4) and calcium gene-related peptide (CGRP) in neurons in dorsal root ganglion (DRG) of rats with neuropathic pain (NP). Methods Forty healthy male Sprague-Dawley rats, weighing 250-280 g, were divided into 2 groups (n=20 each) using a random number table method: solvent group (group S) and group NP.NP was induced by intraperitoneally injecting resiniferatoxin 210 μg/kg, and the solvent of resiniferatoxin was intraperitoneally injected in group S. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured in 5 rats selected before establishing the model and at 1, 3, 7 and 42 days after establishing the model.Five rats were sacrificed at 1, 3, 7 and 42 days after establishing the model, and the L4-6 segments of the DRGs were removed to determine the expression of CGRP and IB4 in neurons using immunofluorescence. Results Compared with the baseline before establishing the model, the MWT was significantly decreased at 3, 7 and 42 days after establishing the model, and the TWL was prolonged at 1, 3, 7 and 42 days after establishing the model in group NP (P<0.05). Compared with group S, the MWT was significantly decreased at 3, 7 and 42 days after establishing the model, the TWL was prolonged at 1, 3, 7 and 42 days after establishing the model, and the expression of IB4 and CGRP in neurons in DRGs was down-regulated at 1, 3, 7 and 42 days after establishing the model in group NP (P<0.05). Conclusion Down-regulated expression of IB4 expression in neurons in DRGs may be involved in the development and maintenance of mechanical hypersensitivity to pain, and down-regulated expression of CGRP may be involved in the development and maintenance of thermal analgesia in rats with NP. Key words: Neuralgia; Ganglia, spinal; Plant lectins; Calcitonin gene-related peptide

The Association of TSHR Gene rs2268458 Polymorphism with Hypothyroidism in Females of Babylon Province-Iraq

The Aim: To evaluate the association of TSHR gene polymorphism rs2268458 with some Physio-biochemical parameters among hypothyroidism women.&#x0D; Methods: This study included 51 hypothyroidism women and 43 healthy women as a control group. Some Physio-biochemical (body mass index (BMI), cytochrome 450 1A1 (CYP1A1), calcium, and phosphorus) and hormonal assay (Osteopontin (OPN), thyroid stimulating hormone (TSH), fT3, fT4 and Calcitonin (CT)) assay were performed. Genotyping of rs-2268458 of TSHR gene was carried out using the polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) technique.&#x0D; Results: TSHR rs2268458 was associated with hypothyroidism in Iraqi women and allele C can be a risk factor for disease by OR= 4.393, CI=1.42-13.53 where TC and CC genotypes were associated with high BMI, while TC genotype was associated with high calcitonin and CYP1A1 levels.&#x0D; Conclusion: The TSHR rs2268458 polymorphism was associated with hypothyroidism in Iraqi women and allele C can be a risk factor for some Physio-biochemical and hormonal disorder in hypothyroidism women.

1827P - Surgery for lymph node metastases of sporadic medullary thyroid carcinoma

Background: Medullary thyroid carcinoma (MTC) arises from malignant proliferation and differentiation of C cells, approximately 2% of all thyroid malignancies. Calcitonin (CT) and carcinoembryonic antigen (CEA) secreted by C cells are the main secretory product and serves as a marker for the diagnosis of this disease. It is also used in the follow-up of MTC patients after thyroidectomy for the identification of relapse or progression of disease. Compared with other thyroid cancers, MTC has unique biological characteristics and is insensitive to radiotherapy, chemotherapy and iodine therapy. Surgical intervention is currently the only effective, curative treatment for medullary thyroid cancer. The 2015 American Thyroid Association recommended total thyroidectomy and different range of lymph node dissection, but controversy remains surrounding the indication for prophylactic lateral lymph node dissection. This study was performed to analyze the risk factors for cervical lymph node metastases and predict the indication for prophylactic lateral neck dissection in patients with sporadic medullary thyroid carcinoma (SMTC). Methods: The aims of this study were to analyze the risk factors of cervical lymph node metastases, and also to predict the indication of prophylactic lateral neck dissection in SMTC patients. Results: Metastases rates in central and lateral compartment were 46.2% (30/65) and 40.0% (26/65), respectively. Univariate analyses showed the incidence of cervical lymph node metastases was significantly higher in patients with tumor size >1cm, tumor multifocality and thyroid capsule invasion. Multivariate analyses revealed that only thyroid capsule invasion was an independent predictive factor for central compartment metastases (p < 0.001, OR = 11.080) and lateral neck metastases (p < 0.001, OR = 9.067). Moreover, the possibility of central compartment metastases was higher when preoperative value of serum carcinoembryonic antigen (CEA) was above 30 ng/mL (60% vs 34.3%, χ2=4.298, P = 0.038). Conclusions: The medullary thyroid carcinoma has a high incidence of cervical lymph node metastases. Prophylactic lateral node dissection is necessary in patients with thyroid capsule invasion or with high value of serum carcinoembryonic antigen (CEA). Funding: Has not received any funding. Disclosure: All authors have declared no conflicts of interest.

Fasting and refeeding induce differential changes in hypothalamic mRNA abundance of appetite-associated factors in 7 day-old Japanese quail, Coturnix japonica

There is little information regarding effects of fasting on feeding behavior and hypothalamic physiology in young Japanese quail. The aim was thus to measure food intake and hypothalamic mRNA in response to fasting and refeeding. Five d-old quail ate little during the dark cycle. Food intake was greatest during the first 2 h of the light cycle. Six day-old quail fasted for 6 h ate the most during the first 15 min of refeeding. In 7 d-old quail, 3 h of fasting up-regulated hypothalamic neuropeptide Y (NPY), NPY receptor sub-type 2 (NPYR2), agouti-related peptide (AgRP), orexin receptor 2 (ORXR2), melanocortin receptors 3 and 4 (MC3R and MC4R, respectively), and neuropeptide S (NPS) and decreased corticotropin-releasing factor receptor sub-type 1 (CRFR1) mRNA. Quail fasted for 3 h and refed for 1 h had greater NPY, AgRP, POMC, and MC3R but less CRFR1 mRNA than fed quail. Quail fasted for 6 h expressed more NPY, NPYR1, NPYR2, and MC3R and less ORXR2, prolactin releasing peptide (PrRP), cocaine- and amphetamine-regulated transcript (CART), and calcitonin (CAL) mRNA than fed quail. Quail fasted for 6 h and refed for 1 h expressed more NPY, NPYR1, NPYR2, AgRP, MC3R, MC4R, and NPS and less galanin, ORXR2, PrRP, CART, and CAL mRNA than fed birds. Hence, fasting induced changes in hypothalamic mRNA, with the largest changes occurring in genes associated with NPY and melanocortin signaling. Most genes remained elevated or downregulated after refeeding, suggesting that there was a time lag for transcription to respond to compensatory feeding.

Analysis of the influence of ureteroscopic pneumatic lithotripsy on the serum levels of CRP, IL-1 and TNF-αin patients with upper ureteral calculi

Objective To investigate the therapeutic effect of ureteroscopic pneumatic lithotripsy (URSL) on ureteral calculi and the effect on serum CRP, IL-1, TNF-αlevels. Methods Retrospective analysis of clinical data of patients with upper ureteral calculi from February 2015 to October 2016 in our hospital was performed. All patients were divided into ureteroscopy lithotripsy group (URSL group) and percutaneous nephrolithotomy group.The general situation of the two groups were observed. The levels of serum inflammatory factors, urinary function and quality of life were compared between the two groups before and after operation. Results The operation time and hematuria time of URSL group were shorter than those of percutaneous nephrolithotomy (t=27.415, 12.018, P<0.001). The stone free rate was higher than that in the percutaneous nephrolithotomy group.The level of inflammatory factors in the two groups had no difference before treatment, after treatment, the level of CRP, TNF-α, NF-κB alpha and IL-1 of the URSL group were lower than those of percutaneous nephrolithotomy group. The IPSS score of patients in URSL group was lower than that of percutaneous nephrolithotomy group at 1 month and 3 months after operation, and Qmax was higher than that of percutaneous nephrolithotomy group.There was no difference in quality of life between the two groups before operation. After operation, the quality of life score of URSL group was higher than that of percutaneous nephrolithotomy group in the 3 months after operation. Conclusions URSL has a good effect on the treatment of upper ureteral calculi, it can reduce postoperative serum CRP, IL-1, TNF-αlevel, im- prove the quality of life. Key words: Ureteral Calculi; Nephrostomy, Percutaneous; Ureteroscopy; Calcitonin Gene-Related Peptide; Interleukin-1; Tumor Necrosis Factor-alpha

Clinical observation of modified Baizhu-Shaoyao powder for the type 2 diabetes with intestinal dysfunctional diarrhea

Objective To explore clinical efficacy and safety of modified Baizhu-Shaoyao powder for treatment of type 2 diabetes complicated with intestinal dysfunctional diarrhea. Methods According to the random indicator method, 109 patients with type 2 diabetes complicated with intestinal dysfunctional diarrhea in our hospital were divided into control group (n=54) and treatment group (n=55). Patients of control group were treated with conventional treatment, while the treatment group were treated with modified Baizhu-Shaoyao powder combined with conventional treatment. The two groups were treated for 14 days. The level of fasting blood glucose (FBG), postprandial 2 h blood glucose(2 hPG), glycosylated hemoglobin (HbAlc) of the two groups before and after treatment was compared by fully automatic biochemical analyzer; The levels of 5-hydroxytryptamine (5-HT), calcitonin gene related peptide (CGRP) and endothelin (ET) was detected by ELISA method. Results The total effective rate of observation group was 94.6% (52/55), which was significantly higher than the control group 70.4% (38/54), and the difference was statistically significant (χ2=11.064, P=0.001). After treatment, the levels of FBG (6.04 ± 1.01 mmol/L vs. 7.36 ± 1.12 mmol/L, t=6.464), 2 hPG (9.05 ± 1.76 mmol/L vs. 10.10 ± 1.92 mmol/L, t=2.977]), HbAlc (7.02% ± 0.23% vs. 7.29% ± 0.50%, t=3.633) in the treatment group were lower than those in the control group (P<0.05). After treatment, the serum levels of 5-HT (20.10 ± 8.22 nmol/L vs. 29.24 ± 9.62 nmol/L, t=5.336), CGRP (51.04 ± 6.76 pmol/L vs. 67.41 ± 7.25 pmol/L, t=12.195), ET (45.16 ± 6.36 pmol/L vs. 52.39 ± 8.44 pmol/L, t=5.057) in the treatment group were lower than those in the control group (P<0.05). There was no significantly difference of the adverse reaction rates of the two groups during treatment (χ2=0.229, P=0.632). Conclusions Modified Baizhu-Shaoyao powder combined with western medicine routine treatmentcan improve the patients’ blood sugar level with type 2 diabetes with intestinal dysfunctional diarrhea, reduce the serum level of 5-HT, CGRP and ET and improve the clinical efficacy. Key words: Diabetes mellitus, type 2; Gastrointestinal tract; Diarrhea; Modified Baishu-Shaoyao powder; Integrated Chinese traditional and western medicine therapy

Anti-inflammatory role of methotrexate in adjuvant arthritis: effect on substance p and calcitonin gene-related Peptide in thymus and spleen.

To investigate the anti-inflammatory effect of methotrexate (MTX) in rats with adjuvant arthritis through its influence on the expression of proinflammatory neuropeptides, substance P (SP) and calcitonin gene-related peptide (CGRP) in immune organs, thymus and spleen. Phase-I pre-clinical trial. The Aga Khan University, Karachi, from July to December 2003. Adjuvant arthritis was induced in rats by inoculation with heat-killed mycobacteria. One group of arthritic rats (n=6) was treated with MTX (0.2 mg/kg body weight, subcutaneously) on every 4th day for a period of 18 weeks, while another group of arthritic rats (n=6) was treated with physiological saline served as control. At the end of experiment, animals were sacrificed and thymus and spleen were dissected and prepared for immunohistochemical analysis. The neuronal density of SP and CGRP immunoreactivity in thymus and spleen was assessed by semi-quantitative analysis. There was a marked reduction in hind paw swelling and inflammation in the MTX-treated rats after 18 weeks of treatment. Restoration of joint spaces (tibiotalar and subtalar) was seen after 9 weeks of MTX treatment. CGRP-positive nerve fibres were significantly reduced (p=0.0001) in thymus of rats treated with MTX compared to control rats. SP-positive nerve fibers were also found to be decreased in thymus of rats treated with MTX compared to controls, however, the decrease was not statistically significant. The neuronal density of SP and CGRP-immunoreactivity in spleen was not significantly different in MTX-treated and placebo-treated rats. In arthritic rats, MTX significantly reduced CGRP expression in thymus. Suppression of pro-inflammatory neuropeptides, such as CGRP and probably SP could be another mechanism by which MTX produces its anti-inflammatory effect in adjuvant arthritis.

Uso della procalcitonina come marker del carcinoma midollare tiroideo

Here we reviewed the literature on the use of procalcitonin (PCT), a precursor of calcitonin (CT), as diagnostic marker of medullary thyroid carcinoma (MTC) in both preoperative and postsurgical setting. To date, about forty articles have been published on this topic. Some of these evaluated the reliability of PCT to detect MTC in thyroid nodule patients, and the other ones investigated the performance of PCT to identify recurrent MTC during postoperative follow-up. Based on published data, PCT measurement appears a promising and reliable serum marker for the diagnosis of MTC. The sample handling of PCT is less laborious and in specific conditions its assay could have significant impact and higher accuracy than CT. Some potential cut-offs have been reported but it would be worthwhile to establish a fixed threshold to use PCT during clinical practice.

The correlation between calcitonin and c-reactive protein and anastomotic fistula after colorectal cancer

The correlation between calcitonin and c-reactive protein and anastomotic fistula after colorectal cancer

Mining and Functional Verification of Calcitonin Genes from the Genome Database.

With more countries in the world entering elderly society, osteoporosis is a common disease among the elderly, especially middle-aged and elderly women. Although calcitonin is an effective drug used to treat osteoporosis in clinical practice, it also exists such problems as high cost, short half-life, and high immunogenicity. Therefore, to explore more efficient calcitonin has important clinical significance. Given the emergence of new-generation gene sequencing, numerous genome sequences of marine species have been revealed. This study aimed to identify new, highly active Calcitonins (CTs) from the gene database. Candidate CT sequences were obtained by BLAST and analyzed. The evolutionary tree of these sequences was constructed using the Neighbor-Joining method of MEGA 7 software. Secondary structures were analyzed by Circular Dichroism (CD). The biological activities of CTs were estimated using the standard of the rat hypocalcemic activity assay in vivo. The half-life and immunogenicity of CT sequences were determined by ELISA. The physicochemical properties of peptides were analyzed with ProtParam and HeliQuest. A total of 64 candidate CT gene and amino acid sequences from different species were obtained by BLAST using the salmon CT (sCT) sequence as the query sequence. These sequences were clustered to 27 different CT polypeptide sequences, and then the evolutionary tree was constructed. A total of 13 sequences were selected for chemical synthesis and activity assay. Results showed that although their secondary structures were similar, four types of candidate CTs had 30% higher activities than sCT, three other types had similar activities to sCT, and the remaining four types had much lower activities than sCT. Among the three designed CTs, the activities of CT-01 and CT-02 were at least 50% higher than those of sCT. Furthermore, all three CT sequences had a similar half-life to sCT and lower immunogenicity. CTs from Monodelphis domestica, Gallus gallus, Ornithorhynchus anatinus, and Carassius auratus had high activities. The exploration and mining of the marine-life genome database can be extremely valuable considering broad application prospect.

Protective effect of myocardial calcitonin gene-related peptide gene transduction on myocardial ischemia/reperfusion injury in diabetic rats

Objective To evaluate the feasibility of lentivirus (Lv) mediated calcitonin gene-related peptide(CGRP) gene transduction in diabetic rats by myocardial injection and whether this method has a protective effect on diabetic myocardium during ischemia/reperfusion(I/R) injury. Methods Sixty healthy male SD rats, weighing 200-220 g, were randomly divided into control group (group C), diabetes mellitus group (group DM), Sham transduction group (group Sham), myocardial injection control group (group MC) and myocardial injection experiment group(group ME), in accordance with a random number table. Rats in group Sham, group MC and group ME were given with normal saline, at a titer of 2×107 virus genomes(vg)/ml Lv-enhanced green fluorescent protein (eGFP) and Lv-CGRP-eGFP 50 μl respectively by myocardial injection. One week later, rats in group DM, group Sham, group MC and group ME received streptozocin(STZ) 50 mg/kg intraperitoneal injection to induce diabetes. Eight weeks after the injection of STZ, myocardial I/R was induced by occlusion and release of left anterior descending branch of coronary artery. The latency of tail flick reflection, the expression of eGFP, the transcription level of CGRP, the expression of CGRP in myocardium, spinal cord and dorsal root ganglion(DRG) and myocardial ischemia area, infarct size(IS) area were measured. Results Compared with group C, the latency of tail flick reflection in group DM, group Sham, group MC and group ME were significantly prolonged in the 4th week after diabetic model successfully manufactured(P<0.05). The transcription levels of CGRP in the ischemic myocardium, spinal cord T1-T5 and DRG T1-T5 in group DM, group Sham and group MC were lower than those in group C and group ME (P<0.05). The level of CGRP in ischemic myocardium and DRG T1-T5 of group ME was higher than that in group C(P<0.05). The content of CGRP in cord T1-T5, DRG T1-T5, ischemic myocardium and serum in group C and group ME were higher than those in group DM, group Sham and group MC (P<0.05). The content of CGRP in cord T1-T5, ischemic myocardium in group ME was higher than that in group C (P<0.05). The infarct size in group DM, group Sham, group MC were higher than that in group C and group ME(P<0.05), the infarct size of group ME was higher than that in group C(P<0.05). Conclusions Intramyocardial injection of CGRP gene can effectively enhance the expression of CGRP at I/R of diabetic rat′ heart. Increased CGRP may reduce cardiac I/R injury in diabetic rats. Key words: Calcitonin gene-related peptide; Diabetes mellitus; Myocardium; Ischemia-reperfusion

Investigation of the relationship between MTHFR, IRS and CALCA gene polymorphisms and development of diabetic nephropathy in patients with type 2 diabetes mellitus

ABSTRACTThe aim of this study was to investigate the relationship between MTHFR, IRS and CALCA gene polymorphisms and development of diabetic nephropathy (DN) in patients with type 2 diabetes mellitus (DM). Our study included 93 patients with type 2 DM diagnosed as having nephropathy and 95 controls diagnosed with type 2 DM without development of DN. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were used to determine the genotype distributions of MTHFR, IRS and CALCA gene polymorphisms. The results showed no statistically significant difference between DN patients and type 2 DM controls in terms of genotype distributions of MTHFR (C677T, A1298C), IRS (IRS-1 Gly972Arg, IRS-2 Gly1057Asp) and CALCA T692C gene polymorphisms (p > 0.05). However, in terms of allele frequencies for the MTHFR A1298C gene, the frequency of the C allele was significantly higher in the DN patients compared to the controls (p < 0.05). In the IRS-2 Gly1057Asp gene polymorphism, the G allele frequency was significantly higher in the DN patients than in the type 2 DM controls (p < 0.05). In the DN group, the individuals with one or less mutant alleles were significantly more than in the control group in terms of the IRS-2 Gly1057Asp gene polymorphism (p < 0.05). The C allele frequency for the MTHFR A1298C gene polymorphism and the G allele frequency for the IRS-2 Gly1057Asp gene polymorphism were indicated to be potential a genetic risk factor for the development of DN in patients with type 2 DM who developed DN.

Serum calcitonin gene-related peptide facilitates adipose tissue lipolysis during exercise via PIPLC/IP3 pathways.

Calcitonin gene-related peptide (CGRP) is formed by alternative transcription of the calcitonin/α-CGRP gene, which also gives rise to calcitonin (CT). Recently, CGRP has been the focus of research for its metabolic effects in vitro. In the present study, the in vivo effects of CGRP on epididymal fat pads lipolysis at rest and during exercise were investigated in trained male Wistar rats. Male Wistar rats were assigned to control and trained groups, which underwent endurance training for 12 weeks. The control (at rest) and trained (during acute exercise) animals were subjected to an intravenous injection of rat recombinant CGRP (2 µg kg-1) and CGRP-(8-37), a competitive CGRP receptors antagonist, to evaluate if and how CGRP can affect adipose tissue lipolysis at rest and during exercise. Intravenous injection of rat CGRP recombinant at rest upregulated major lipolysis pathways (cyclic AMP (cAMP), AMP-activated protein kinase (AMPK), and phospholipase C (PIPLC/IP3)) in fat pads, causing an elevation in plasma-free fatty acid (FFA) and a decrease in plasma triglyceride (TG). All the effects were eliminated by pretreating the animals with CGRP-(8-37), suggesting that CGRP receptors were necessary for lipolytic effects of CGRP in fat pads. In trained animals, acute exercise augmented CGRP in serum, cerebrospinal fluid (CSF), and the cortex. Pretreating the animals with CGRP-(8-37) attenuated PIPLC/IP3 pathway in fat pads and had no effect on cAMP and AMPK pathways. Epididymal fat pads is a metabolic target for CGRP during exercise and CGRP effects on adipose tissue metabolism during exercise could be related to PIPLC/IP3 pathway.

Biochemical, Anatomical, and Pharmacological Characterization of Calcitonin-Type Neuropeptides in Starfish: Discovery of an Ancient Role as Muscle Relaxants.

Calcitonin (CT) is a peptide hormone released by the thyroid gland that regulates blood Ca2+ levels in mammals. The CT gene is alternatively spliced, with one transcript encoding CT and another transcript encoding the CT-like neuropeptide calcitonin-gene related peptide (α-CGRP), which is a powerful vasodilator. Other CT-related peptides in vertebrates include adrenomedullin, amylin, and intermedin, which also act as smooth muscle relaxants. The evolutionary origin of CT-type peptides has been traced to the bilaterian common ancestor of protostomes and deuterostomes and a CT-like peptide (DH31) has been identified as a diuretic hormone in some insect species. However, little is known about the physiological roles of CT-type peptides in other invertebrates. Here we characterized a CT-type neuropeptide in a deuterostomian invertebrate—the starfish Asterias rubens (Phylum Echinodermata). A CT-type precursor cDNA (ArCTP) was sequenced and the predicted structure of the peptide (ArCT) derived from ArCTP was confirmed using mass spectrometry. The distribution of ArCTP mRNA and the ArCT peptide was investigated using in situ hybridization and immunohistochemistry, respectively, revealing stained cells/processes in the nervous system, digestive system, and muscular organs, including the apical muscle and tube feet. Investigation of the effects of synthetic ArCT on in vitro preparations of the apical muscle and tube feet revealed that it acts as a relaxant, causing dose-dependent reversal of acetylcholine-induced contraction. Furthermore, a muscle relaxant present in whole-animal extracts of another starfish species, Patiria pectinifera, was identified as an ortholog of ArCT and named PpCT. Consistent with the expression pattern of ArCTP in A. rubens, RT-qPCR revealed that in P. pectinifera the PpCT precursor transcript is more abundant in the radial nerve cords than in other tissues/organs analyzed. In conclusion, our findings indicate that the physiological action of CT-related peptides as muscle relaxants in vertebrates may reflect an evolutionarily ancient role of CT-type neuropeptides that can be traced back to the common ancestor of deuterostomes.

Diagnostic efficacy of inflammatory markers in maternal and umbilical cord blood in histologic chorioamnionitis

Objective To explore the clinical value of maternal and umbilical blood inflammatory markers, interleukin-6 (IL-6), neutrophil-to-lymphocyte ratio (NLR), C-reactive protein (CRP) and pro-calcitonin (PCT), in the diagnosis of histologic chorioamnionitis (HCA). Methods A total of 102 suspected chorioamnionitic cases were enrolled from January 2014 to July 2017. They were assigned into two groups based upon postpartum histopathological examination of placenta: HCA group (48 cases) and control group (54 cases). Maternal and umbilical blood samples were collected for routine blood test and tested for IL-6, NLR, CRP and PCT levels. T, Mann-Whitney U or Chi-square (or Fisher's exact) test was used for data comparison. Meaningful indicators in maternal and umbilical cord blood were analyzed by logistic regression analysis and correlation analysis. At the same time, receiver operating characteristic (ROC) curve was drawn to evaluate their diagnostic values. Results (1) IL-6 level and NLR in maternal blood in HCA group were higher than those in control group [6.95 (2.40-13.50) vs 3.90 (2.30-9.20) pg/ml, Z=-5.147; 5.03 (1.92-9.20) vs 3.94 (1.85-11.17), Z=-3.097; both P<0.05], and the levels of white blood cells, neutrophile granulocytes, CRP and IL-6 as well as NLR in umbilical cord blood were also higher [(9.4±2.0)×109/L vs (8.6±1.4)×109/L, t=-2.522; (6.87±1.62)×109/L vs (5.99±1.26)×109/L, t=-3.071; 12.30 (0.50-89.04) vs 3.18 (0.50-88.93) mg/L, Z=-4.519; (8.78±2.56) vs (4.78±1.45) pg/ml, t=-7.025; (4.45±1.36) vs (3.78±1.22), t=-3.020; all P<0.05]. (2) Logistic regression analysis showed that elevated levels of IL-6 and NLR in maternal blood and CRP and IL-6 in umbilical cord blood were independent risk factors for HCA [OR (95%CI): 1.65 (1.32-2.06), 1.34 (1.02-1.77), 1.05 (1.00-1.11) and 2.39 (1.72-3.32), all P<0.05]. Positive correlations were found between the levels of IL-6 in maternal and umbilical cord blood, and between NLR in maternal blood and CRP level in umbilical cord blood (correlation coefficient: 0.680 and 0.230, both P<0.05). (3) IL-6 level in umbilical blood was of the greatest value in the diagnosis of HCA among all single markers, followed by IL-6 in maternal blood, CRP in umbilical blood and NLR in maternal blood [area under the ROC curve (AUC): 0.904, 0.796, 0.760 and 0.678]. When two indexes were combined, NLR in maternal blood+IL-6 in umbilical cord blood showed the highest diagnostic value, followed by, IL-6 in maternal blood+CRP in umbilical cord blood, IL-6+NLR in maternal blood and NLR in maternal blood+CRP in umbilical cord blood (AUC: 0.917, 0.870, 0.823 and 0.791). When three indexes was used in combination, the diagnostic value of IL-6 in maternal and umbilical cord blood+NLR in maternal blood was higher than that of IL-6 and NLR in maternal blood+CRP in umbilical cord blood (AUC: 0.919 and 0.836). (4) There were 13 cases (27.1%) with neonatal complications in HCA group and two (3.7%) in control group (P<0.05). Conclusions Changes in NLR and IL-6 levels in maternal blood and NLR, IL-6 and CRP levels, and white blood cells and neutrophile granulocytes counts in umbilical cord blood are associated with HCA. The diagnostic efficacy of two indexes combined is superior to that of single index, while the combination of three indexes can significantly improve the diagnostic accuracy and authenticity. Key words: Chorioamnionitis; Interleukin-6; C-reactive protein; Calcitonin; Leukocyte count