Effects of tegaserod on Fos, substance P and calcitonin gene-related peptide expression induced by colon inflammation in lumbarsacral spinal cord

AIM: To investigate the mechanisms of tegaserod, a partial 5-HT4 agonist, in reducing visceral sensitivity by observing Fos, substance P (SP) and calcitonin gene-related peptide (CGRP) expression in the lumbarsacral spinal cord induced by colonic inflammation in rats. METHODS: Twenty-four male rats with colonic inflammation induced by intraluminal instillation of trinitrobenzenesulfonic acid (TNBS) were divided into 3 groups. Treatment group 1: intra-gastric administration of tegaserod, 2 mg/kg.d; Treatment group 2: intra-gastric administration of tegaserod, 1mg/kg.d; Control group: intra-gastric administration of saline, 2.0 mL/d. After 7 d of intra-gastric administration, lumbarsacral spinal cord was removed and processed for Fos, SP and CGRP immunohistochemistry. RESULTS: In rats of the control group, the majority of Fos labeled neurons was localized in deeper laminae of the lumbarsacral spinal cord (L5-S1). SP and CGRP were primarily expressed in the superficial laminae of the spinal cord after TNBS injection. Intra-gastric administration of tegaserod (2 mg/kg.d) resulted in a significant decrease of Fos labeled neurons (22.0±7.7) and SP density (12.5±1.4) in the dorsal horn in the lumbarsacral spinal cord compared to those of the control group (62.2±18.9, 35.9±8.9, P<0.05). However, CGRP content in dorsal horn did not significantly reduce in rats of treatment group 1 (1.2+1.1) compared to that of the control group (2.8±2.4,P>0.05). Neither Fos expression nor SP or CGRP density in the dorsal horn significantly declined in rats of treatment group 2 compared to those of the control group (P>0.05). CONCLUSION: Tegaserod can significantly reduce Fos labeled neurons in the lumbarsacral spinal cord induced by colonic inflammation. Tegaserod may reduce visceral sensitivity by inhibiting SP expression in the dorsal horn of spinal cord.

Levels of vasoactive intestinal peptide,cholecystokinin and calcitonin gene-related peptide in plasma and jejunum of rats following traumatic brain injury and underlying significance in gastrointestinal dysfunction

AIM:To study the alterations of brain-gut peptides following traumatic brain injury (TBI) and to explore the underlying significance of these peptides in the complicated gastrointestinal dysfunction.METHODS:Rat models of focal traumatic brain injury were established by impact insult method,and divided into 6 groups (6 rats each group) including control group with sham operation and TBI groups at postinjury 3,12,24,72h,and d 7.Blood and proximal jejunum samples were taken at time point of each group and gross observations of gastrointestinal pathology were recorded simultaneously.The levels of vasoactive intestinal peptide (VIP) in plasma,calcitonin gene-related peptide (CGRP) and cholecystokinin (CCK) in both plasma and jejunum were measured by enzyme immunoassay (EIA). Radioimmunoassay (RIA) was used to determine the levels of VIP in jejunum.RESULTS:Gastric distension,delayed gastric emptying and intestinal dilatation with a large amount of yellowish effusion and thin edematous wall were found in TBI rats through 12h and 72h, which peaked at postinjury 72h. As compared with that of control group (247.8±29.5ng/L), plasma VIP levels were significantly decreased at postinjury 3,12 and 24h (106.7±34.1ng/L, 148.7±22.8ng/L,132.8±21.6ng/L,respectively),but significantly increased at 72h (405.0±29.8ng/L) and markedly declined on d 7 (130.7±19.3ng/L).However,Plasma levels CCK and CGRP were significantly increased through 3h and 7d following TBI (126-691% increases),with the peak at 72 h.Compared with control (VIP, 13.6±1.4ng/g;CGRP,70.6±17.7ng/g);VIP and CGRP levels in jejunum were significantly increased at 3h after TBI (VIP,35.4±5.0ng/g;CGRP,103.8±22.1ng/g),and declined gradually at 12 h and 2d h (VIP,16.5±1.8ng/g,5.5±1.4ng/g;CGRP,34.9±9.7ng/g, 18.5±7.7ng/g),but were significantly increased again at 72 h (VIP, 48.7±9.5ng/g; CGRP,142.1±24.3ng/g),then declined in various degrees on d 7 (VIP, 3.8±1.1ng/g; CGRP, 102.5±18.1ng/g).The CCK levels in jejunum were found to change in a similar trend as that in plasma with the concentrations of CCK significantly increased following TBI (99-517% increases) and peaked at 72h.CONCLUSION:Traumatic brain injury can lead to significant changes of brain-gut peptides in both plasma and small intestine, which may be involved in the pathogenesis of complicated gastrointestinal dysfunction.

Study of Calcitonin and Calcitonin Gene-related Peptide in Patients with Medullary Thyroid Carcinoma

OBJECTIVE To provide a basis for treatment and prognosis for MTC patients by investigating CT and CGRP.METHODS Fifty-eight cases of MTC were selected and the relationship between the CT levels and metastasis was investigated. An immunohistochemical method was used to detect the expression of CT and CGRP in the 58 samples of MTC tissues. The CT and CGRP of new MTC inpatients were measured before operation and in the first few days after operation using a radioimmunoassy.RESULTS 1) The metastatic rate of cervical lymph nodes was a significantly different between the normal serum CT group and the elevated group (P<0.01). 2) The rate of residual tumor was significantly different between the normal serum CT group one month after operation and the elevated group at the same period (P<0.01). 3) In the immunohistochemical study we found the positive rate of CT was about 98%, and the CGRP was 87.8%. 4) Part of the patients had an elevated CGRP level while CT levels were normal. 5) The serum CT levels dropped to a stable range one week after operation.CONCLUSION The CT levels are a useful index to evaluate the probability of cervical lymph node metastasis and the efficacy of surgical treatment.Measurement of the serum CGRP is an aid for the diagnosis of MTC,especially for those patients whose preoperative CT levels are normal.

Investigation of Calcitonin and Calcitonin Gene-related Peptide in 88 Cases of Medullary Thyroid Carcinoma

Objective： To investigate the changes of calcitonin （CT） and calcitonin gene-related peptide （CGRP） in patients with medullary thyroid carcinoma （MTC）. Methods： Fifty-eight cases of MTC were selected and the relationship between the CT levels and metastasis was investigated. The immunohistochemical method was used to detect the expression of CT and CGRP in the 58 samples of MTC tissues. The CT and CGRP in 30 newly diagnosed MTC inpatients were measured before operation and in the first few days after operation using a radioimmunoassy. Results： （1） The rate of residual tumor had a significant difference between the normal serum CT group one month after operation and the elevated group at the same period （P〈0.01）. （2） Immunohistochemical study revealed the positive rate of CT was about 98%, and that of the CGRP was 87.8%. （3） Part of the patients had an elevated CGRP levels while CT levels was normal. （4） The serum CT levels were decreased to a stable range one week after operation. Conclusion： CT is a useful index to evaluate the efficacy of surgical treatment. The measurement of serum CGRP is helpful in the diagnosis of MTC, especially for those whose preoperative CT levels are normal.

Effect of Acupuncture on Calcitonin in Ovariectomized Rats

Objective To Explore the mechanism of the acupuncture on posanenopausal osteoporo-sis.Method One hundred Wistar female rats(6 month- old)were randomly divided into two groups: O-variectomized group and sham - operated group. After 3 months of operation, three rats were chosen tomeasure the bone histomorphometric indexes. Then the ovariectomized rats were distributed randomly to acupuncture group and yigaining group and normal saline group and control 2 group, the rats were respec-tively treated by acupuncture and yigaining and normal saline. After the treatment for three course, all rats were collected blood from the ateria femoralis, the levels of calcitonin were measured by IRMA. the bone histomorphometric indexes and C - cell in thyroid follicle were detected. Results The levels of calcitonin of acupuncture group were increasing significantly comparing with the yigaining group and the normal saline group and the control 2 group ( p < 0.05), but there were no evident difference in the levels of cal-citonin between the acupuncture group and the sham - operated group. The mean trabecular bone width and the tmbecuhar bone volume % of acupuncture group were increasing more than that of the normal saline group and the control 2 group(p < 0.05) ;the mean marrow cavity width and the marrow cavity vol-ume % were derreasing more than that of the nomal saline group and the control 2 group( P < 0.05) ; the number of C - cell in thyroid follicle of the acupuncture group were increasing more than that of the other groups except the sham - operated group. Conclusion Acupuncture can effectively treat the post-menopausal osteoporosis by strengthening the ability of the C - cell in thyroid follicle which can improve the levels of calcitonin in serum.

Bone tumor

＂T＂ shape autologous bone transplantation in treating giant cell tumor of bone around the knce; Methods and opportunity of operation for the malignant tumor intercurrent fracture of extremity;Limb salvage methods of bone tumors around the knee joint;Prevention and treatment of the aseptic loosening of prosthesis for the rabbits of osteoporosis by the salmon calcitonin： an experimental study;……

Others

Prevention and treatment of the aseptic loosening of prosthesis for the rabbits of osteoporosis by the salmon calcitonin： an experimental study; Preliminary observation of stem cells and olfactory ensheathing cells garaft for treatment of spinal cord injury;The study of tissue-engineering bone for repair of segmental bone defects; Immunologic properties of undifferentiated messenchymal stem cells;Construction of recombinant adenovirus vector containing human vascular endothelial growth factor and its expression in mesenchymal stem cells; Mechanism of alendronate preventing the periprostetical osteolysis by the wear particles;The study on the biodegradation mechanism of the HA/PLGA composite made by high-pressure for internal fixation of bone fractures;An experimental study on protective effect of extracellular ATP to motor neurons in anterior horn of spinal cord after sciatic nerve injury;……

Others——Single nudeotide polymorphism of calcitonin receptor gene and idiopathic hypercalciuria

Objective To explore the association of the calcitonin receptor （CTR） allelic polymorphism in the 1 377 bp region with the risk of idiopathic hypercalciuria （IH） in the Han nationality in Hubei area, and to study the pathogenesis of IH. Methods The CTR genotypes were determined by polymerase chain reaction-restriction flagment length polymorphism in 76 patients with IH and 126 healthy controls from the Han nationality in Hubei area, using restriction endonuclease AluI. Results The distri bution frequencies of AluI alleles in the 2 groups followed the Hardy-Weinberg equilibrium. The distribution frequencies of the CC, TC and TT genotypes were 73.7%, 17.1% and 9.2% in IH patient group, and 89.7%, 9.5 % and 0.8 % in control group; the distribution frequencies of C and T alleles in the 2 groups were 84.2 %, 15.8 % and 94.4 %, 5.6 %, respectively. The distribution frequencies of T and TT alleles were higher, while those of C and CC alleles were lower, compared with control group;the differences between the 2 groups were significant （P 〈 0.05）. Conclusion The results indicate that the C/T single nucleotide polymorphism in the CTR gene play a significant role in the mechanism of IH in the Han nationality in Hubei area in China. 7 refs,2 figs,2 tabs.

Effects of oral administration recombinant Saccharomyces cerevisiae expressing human-salmon calcitonin on hypercalcemia rats~

Recombinant human-salmon calcxtonin （hsCT） was synthesized and expressea on the cell surtaces ot recombinant Saccharomyces cerevisiae （yAGA2-hsCT） using the pAGA2-hsCT expression vector. Expression of recombinant hsCT was verified by flow cytometer.The resorption activity of osteoclasts was inhibited by recombinant hsCT protein which released from the cell surface of yAGA2-hsCT. Blood calcium determination in rats showed the activity of yAGA2-hsCT following oral administration of 5 mg/kg lyophilized transformant yeast to hypercalcemia rats decreases the serum calcium from 2.82 ±0.023 mM to 2.662 ± 0.012 mM.

EFFECT OF ACUPUNCTURE OF "ZUSANLI" AND "WEISHU" ACUPOINTS ON EXPERIMENTAL GASTRIC ULCER

Objective: To observe the therapeutic effect of electroacupuncture (EA) of "Zusanli"(ST 36) and "Weishu"(BL 21)on the experimental gastric ulcer in Xu period of a day for analyzing the best opportunity of acupuncture treatment of gastric ulcer. Methods:49 Wistar rats were randomly divided into ① control group, ② ST 36 Xu group, ③ BL 21 Xu group, ④ ST 36 Chen group, ⑤ BL 21 Chen group, ⑥ ST 36 IT group, and ⑦ BL 21 IT group, with 7 rats in each group. Rat gastric ulcer model was established using intra gastric administration of glacial acetic acid. Gastric mucosal thickness,mucosal muscular defect width,superficial mucosal mucus index (MI),intramucosal MI ,Total MI,plasma 6 keto PGF1α ,serum NO and gastric tissue CGRP concentrations were used as the indexes. The EA treatment was conducted once daily at Xu period (7-9 o’clock in the evening), Chen period (8-10 o’clock in the morning) and irregular time (IT) with 6 days being a therapeutic course, two courses altogether. Results:After EA treatment,①the regenerated mucosal thickness values of the aforementioned 6 EA groups, particularly ST 36 Xu group and BL 21 Xu group, were all apparently higher than that of control group (P<0.05-0.01), while the mucosal muscle layer defect width values of different EA groups, particularly ST 36 Xu group and BL 21 Xu group, were strikingly lower than that of control group (P<0.05-0.01); ②the 3 MI of the 6 EA groups, particularly those of ST 36 Xu group were all significantly higher than those of control group (P<0.05-0.01); ③plasma 6 keto PGF1α contents of the 6 EA groups, particularly those of ST 36 Xu and BL 21 Xu groups, were all significantly higher than that of control group (P<0.05-0.01); ④serum NO and gastric tissue CGRP concentrations of the 6 EA groups, particularly those of ST 36 Xu and BL 21 Xu groups and CGRP concentration of ST 36 Chen group, were all significantly higher than those of control group (P<0.05-0.01);and ⑤there were positive correlation between superficial MI or total MI and NO level (r=0.858, 0.987, P<0.05,and 0.01),and between the superficial mucus index and CGRP concentration (r=0.9051, t=4.68, P<0.01), suggesting that the increase of the secretion function of the regenerated mucosal mucus is related to acupuncture induced increase of release of both NO and CGRP.Conclusion:The therapeutic effect of EA of ST 36 and BL 21 in Xu period is better in the treatment of gastric ulcer,and that of EA of ST 36 is the best.

健康公告栏

法研究人员:加工的面包和谷类食品是造成近视眼主因

Electroacupuncture for functional dyspepsia and the influence on serum Ghrelin, CGRP and GLP-1 levels

Objective： To observe the clinical efficacy of electroacupuncture for functional dyspepsia（FD）, and explore the corresponding mechanism.Methods： Sixty-four FD patients were randomly divided into electroacupuncture group and western medicine group, with 32 cases in each group. In electroacupuncture group, electroacupuncture at Zusanli（足三里ST 36）,Sanyinjiao（三阴交SP 6）,Gongsun（公孙SP 4） and Neiguan（内关PC 6） was performed for once a day, and the needles were retained for 30 min. In western medicine group, oral administration of mosapride citrate dispersible tablets in a dosage of 5 mg/time was carried out for 3 times a day. Treatment was conducted for 30 consecutive days in both groups. The scores of Leeds dyspepsia questionnaire（LDQ） and functional digestive disorder quality of life（FDDQL） of patients in both groups were recorded before and after treatment. Serum Ghrelin, CGRP and GLP-1 levels of patients were tested before and after treatment respectively, and the clinical efficacy of patients in both groups was evaluated after treatment.Results： In western medicine group, LDQ score after treatment was lower than that before treatment（P 0.05）, FDDQL score after treatment was higher than that before treatment, while the differences were not statistically significant（P0.05）. LDQ score in electroacupuncture group after treatment was lower than that before treatment（P0.05）, and also lower than that in western medicine group at the same time point（P 0.05）. FDDQL score in electroacupuncture group after treatment was higher than that before treatment（P0.05）, and also higher than that in western medicine group at the same time point（P0.05）. In western medicine group, Ghrelin level after treatment was higher than that before treatment（P 0.05）, CGRP level reduced, and the differences were not statistically significant（P 0.05）. GLP-1 level after treatment was also higher than that before treatment（P0.05）. In electroacupuncture group,Ghrelin level after treatment was higher than that before treatment, CGRP level reduced, and GLP-1 level after treatment was also higher than that before treatment（both P 0.05）. According to the comparison of values of each index between electroacupuncture group and western medicine group after treatment,the differences were all statistically significant（all P 0.05）. The total effective rate in electroacupuncture group was 90.63%（29/32） which was higher than that in western medicine group 68.75%（22/32）, and the differences were statistically significant（P0.05）.Conclusion： Electroacupuncture at ST 36, SP 6, SP 4 and PC 6 can effectively improve the clinical symptoms of FD patients, and the mechanism might be related with the increase of serum Ghrelin and GLP-1 levels and the decrease of serum CGRP level.

Intermedin/adrenomedullin2: an autocrine/paracrine factor in vascular homeostasis and disease

Intermedin(IMD)or adrenomedullin 2 is a novel peptide related to the calcitonin gene-related peptide(CGRP)family.Via calcitonin receptor-like receptor/receptor activity modifying proteins,the common receptor complexes of CGRP,IMD exerts a wide range of biological effects,especially regulation of cardiovascular homeostasis.Proteolytic processing of a larger IMD precursor yields a series of biologically active C-terminal fragments,IMD1–53,IMD1–47 and IMD8–47.IMD and its receptors are present in the cardiovascular system,and IMD is present at low levels in plasma.In the cardiovascular system,IMD has multiple functions such as regulation of blood pressure and cardiac function,pro-angiogenesis,endothelial barrier function protection,anti-oxidative stress,and anti-endoplasmic reticulum stress.IMD participates widely in the pathogenesis of atherosclerosis,hypertension,pulmonary arterial hypertension and vascular calcification.It is a vascular regulatory factor of homeostasis and a vital endogenous protective factor against vascular diseases.

Modification at Lys24 with fatty acid generates active calcitonin gene-related peptide with much improved plasma stability

As a 37-amino acid vasoactive neuropeptide, calcitonin gene-related peptide （CGRP） is widely distributed in nervous systems. The studies and clinical applications of CGRP are limited by its peptide nature and short half-life. A series of peptide analogues of the α-form of CGRP were synthesized. Afterwards, by using in vitro metabolic and activity studies, we prepared two high affinity analogues with significantly improved plasma stability.