降钙素基因相关肽对卵母细胞体外成熟的影响

目的:探讨降钙素基因相关肽(calcitonin gene-related peptide, CGRP) 对小鼠卵母细胞体外成熟的影响。方法:6 周龄雌性昆明小鼠腹腔注射孕马血清促性腺激素(pregnant mare serum gonadotropin, PMSG)10 U 48 h 后处死, 取出卵巢收集卵母细胞以30~40 枚/ 孔的密度接种于培养板内, 以不同浓度CGRP(0, 10^-10, 10^-9, 10^-8 mol/L) 处理24 h后检测生发泡破裂(germinal vesicle breakdown, GVBD) 率和第一极体(polarbody I, PBI) 排出率;收集、培养人卵巢颗粒细胞, 以不同浓度CGRP(0, 10^-10, 10^-9, 10^-8 mol/L) 处理24 h 后测定颗粒细胞内cAMP 浓度。结果:外源性CGRP处理小鼠卵母细胞可浓度依赖性地降低GVBD 率及PBI 排出率;外源性CGRP 处理人颗粒细胞, 可浓度依赖性地增高细胞内cAMP 浓度。结论:CGRP 抑制小鼠卵母细胞成熟可能与其增加颗粒细胞内cAMP 浓度有关。     

肿瘤坏死因子对体外培养三叉神经节细胞的影响

目的 体外培养完整三叉神经节(TG)组织,建立一种能够易于控制的类似于TG的体内活性体系.探讨肿瘤坏死因子(TNF)-α对体外培养TG细胞中降钙素基因相关肽(CGRP)的影响.方法 将SD大鼠麻醉后取双侧TG,将一部分样本用4%多聚甲醛固定,然后通过HE染色观察其与体外培养TG的神经元之间形态学差异.其余的TG分为常规培养组和添加TNF-α培养组,再通过免疫组化和Western blot观察常规培养组和添加TNF-α培养组TG细胞中CGRP的水平变化,分析TNF-α对体外培养TG细胞的影响.结果 与新鲜分离组相比,体外培养的TG神经元细胞未发生明显的组织细胞水肿、变性、坏死和细胞凋亡等病理学改变.TNF-α培养组与常规培养组相比,TNF-α可使CGRP表达水平上调,且CGRP阳性反应细胞数表达水平显著增高,差异有统计学意义(P<0.05).结论 细胞形态学分析结果显示离体培养的TG组织方法可行,且易于实施形态学和分子生物学研究;TNF-α可使离体培养的TG细胞CGRP表达水平显著增加.     

Effect of pilocarpine on substance P and calcitonin gene‐related peptide releases correlate with salivary secretion in human saliva and plasma

What is known and Objective:  Pilocarpine, a muscarinic receptor agonist, has been used for the treatment of dry mouth. Salivary glands are supplied with nerve fibres that contain neuropeptides, such as substance P, calcitonin gene‐related peptide (CGRP) and vasoactive intestinal polypeptide (VIP), which are important modulators of salivation. It is known that measurement of salivary and plasma levels of neuropeptides is useful for assessing the dose–pharmacological effect relationship of drugs. The relationship between the action of pilocarpine and neuropeptides in humans has not been studied. Moreover, studies evaluate the usefulness of drug salivary levels in the pharmacological evaluation of drugs are scarce. The aim of this study was to examine the effects of pilocarpine on the levels of substance P‐, CGRP‐ and VIP‐like immunoreactive substances (IS) in saliva and plasma taken in healthy humans. Methods:  Five healthy male subjects participated in this study. Pilocarpine tablet (10 mg) or placebo tablet was orally administered with 100 mL of water. Each subject was administered placebo and drug with an interval of 4 weeks in between. Saliva was sampled before and at 20, 40, 60, 90, 120, 180 and 240 min after administration of the test substances. Venous blood samples (10 mL) were also taken from a forearm vein at each time interval. The samples were then enzyme immunoassayed using a highly sensitive system for substance P‐, CGRP‐ and VIP‐IS. The amount of saliva was measured by the Saxon test. Results:  A single oral administration of pilocarpine increased the release of salivary substance P‐IS (the area under the concentration‐time curve: AUC_{0→240 min}) compared with the placebo. Pilocarpine also significantly increased the release of salivary CGRP‐IS (AUC_{0→240 min}). Pilocarpine significantly increased the release of plasma CGRP‐IS. The salivary volume correlated with the salivary level of substance P and CGRP‐IS (r = 0·84, P < 0·05 and r = 0·59, P < 0·05, respectively). AUC_{0→240 min} for substance P‐IS in saliva correlated with that for plasma (r = 0·78, P < 0·05). What is new and Conclusion:  Pilocarpine increases the release of salivary substance P and CGRP‐IS. This suggests that one mechanism by which pilocarpine improves dry mouth is by local stimulation of neuropeptidergic nerves. Moreover, saliva levels of substance P showed good correlation with the plasma levels. The substance P levels in saliva and plasma may be good indicators of the effects of drugs used in dry mouth/xerostomic patients.     

A protocol for angiographic embolization in exsanguinating pelvic trauma: A report on 31 patients

Background?The indication for acquiring angiographic embolization in the initial treatment of severe pelvic fractures is controversial. We describe the characteristics and outcome of 31 patients with traumatic pelvic bleeding who underwent percutaneous angiography with embolization according to a standardized protocol.

Patients and methods?During an 8.5-year period, 1,260 patients were treated for pelvic trauma. We performed a prospective registration of the 46 patients who underwent angiography, and report the 31 patients who had signs of significant arterial injury on angiography, necessitating embolization.

Results?The rate of significant arterial injury after pelvic trauma was 2.5%. All patients had been subjected to high-energy injuries and all were severely injured as measured by the Injury Severity Score: 41 (17–66). Pelvic arterial injury was observed with all types of pelvic trauma, including isolated acetabular (4/31) and sacral fractures (3/31). The internal iliac artery or its branches was injured in 28 of 31 patients. Survival rate after embolization was 84%, and correlated inversely with increasing patient age. None of the patients died of bleeding.

Interpretation?Our findings show that significant pelvic arterial injuries occur in a minority of patients after pelvic trauma, and predominantly affect patients with multiple high-energy injuries regardless of fracture type. The effect of angiographic embolization was good.     

Attenuation of cartilage degeneration by calcitonin gene‐related paptide receptor antagonist via inhibition of subchondral bone sclerosis in osteoarthritis mice

Osteoarthritis (OA) is a progressive joint disorder which affects cartilage and subchondral bone. Calcitonin gene‐related peptide (CGRP) plays a role in bone metabolism. The purpose of this study is to examine the therapeutic effect of the blocking CGRP on OA progression in mice by inhibition of subchondral bone sclerosis. OA was induced by the resection of the medial meniscotibial ligament of the knee in C57/BL6 mice. An intraperitoneal injection of the CGRP receptor antagonist (BIBN4096) was administered after OA surgery. At 1, 4, and 8 weeks after injection, histological analysis were performed. In vitro, the effect of CGRP and BIBN4096 on osteogenesis and osteoclastogenesis was analyzed. BIBN4096 could prevent cartilage degeneration and subchondral bone sclerosis. The OARSI score in the BIBN4096 group was significantly lower than that in the control. In vitro, CGRP up regulated osteocalcin expression, but its expression was down regulated by BIBN4096. CGRP inhibited osteoclastogenesis of raw 267.4 cells, but its effect was reduced by the addition of BIBN4096.The current study showed that subchondral bone sclerosis and increasing expression of CGRP occurs in the early phase of OA in relation to cartilage degeneration, and that BIBN4096 could effectively attenuate OA progression. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1177–1184, 2016.     

围手术期血浆神经肽Y、神经降压素、内皮素-1、降钙素基因相关肽水平的变化

选择胃肠癌患者,分别取术前、术中、术后第１、７天静脑海因,用放射免疫分析测定血浆神经肽Ｙ（ＮＰＹ）神经降压素（ＮＴ）、内皮素－１（ＥＴ－１）、降钙素基因相关肽（ＣＧＲＰ）浓度。结果表明：ＮＹＰ浓度,术中高于术前（Ｐ〈０．０１）,术后第１、７均低于术前（Ｐ〈０．０１）。     

Moderate Ingestion of Alcohol Is Associated With Acute Ethanol‐Induced Suppression of Circulating CTX in a PTH‐Independent Fashion

The “J shape” curve linking the risk of poor bone health to alcohol intake is now well recognized from epidemiological studies. Ethanol and nonethanol components of alcoholic beverages could influence bone remodeling. However, in the absence of a solid underlying mechanism, the positive association between moderate alcoholic intake and BMD remains questionable because of confounding associated social factors. The objective of this work was to characterize the short‐term effects of moderate alcohol consumption on circulating bone markers, especially those involved in bone resorption. Two sequential blood‐sampling studies were undertaken in fasted healthy volunteers (age, 20–47 yr) over a 6‐h period using beer of different alcohol levels (<0.05–4.6%), solutions of ethanol or orthosilicic acid (two major components of beer), and water ± calcium chloride (positive and negative controls, respectively). Study 1 (24 subjects) assessed the effects of the different solutions, whereas study 2 (26 subjects) focused on ethanol/beer dose. Using all data in a “mixed effect model,” we identified the contributions of the individual components of beer, namely ethanol, energy, low‐dose calcium, and high‐dose orthosilicic acid, on acute bone resorption. Markers of bone formation were unchanged throughout the study for all solutions investigated. In contrast, the bone resorption marker, serum carboxy terminal telopeptide of type I collagen (CTX), was significantly reduced after ingestion of a 0.6 liters of ethanol solution (>2% ethanol; p ≤ 0.01, RM‐ANOVA), 0.6 liters of beer (<0.05–4.6% ethanol; p < 0.02), or a solution of calcium (180 mg calcium; p < 0.001), but only after calcium ingestion was the reduction in CTX preceded by a significant fall in serum PTH (p < 0.001). Orthosilicic acid had no acute effect. Similar reductions in CTX, from baseline, were measured in urine after ingestion of the test solutions; however, the biological variability in urine CTX was greater compared with serum CTX. Modeling indicated that the major, acute suppressive effects of moderate beer ingestion (0.6 liters) on CTX were caused by energy intake in the early phase (～0–3 h) and a “nonenergy” ethanol component in the later phase (～3 to >6 h). The early effect on bone resorption is well described after the intake of energy, mediated by glucagon‐like peptide‐2, but the late effect of moderate alcohol ingestion is novel, seems to be ethanol specific, and is mediated in a non–calcitonin‐ and a non–PTH‐dependent fashion, thus providing a mechanism for the positive association between moderate alcohol ingestion and BMD.     

CD64、降钙素原检测在肝硬化自发性细菌性腹膜炎诊断中的意义

目的：检测肝硬化患者 CD64、血清降钙素原（PCT ）水平,探讨 CD64、血清 PCT 检测对自发性细菌性腹膜炎（SBP）的诊断价值。方法将肝硬化患者分为 SBP 组（45例）、腹水非 SBP 组（93例）及对照组3组（50例）,应用流式细胞术测定 CD64;电化学发光免疫法测定 PCT 。分别采集 SBP 组患者在入院后24 h 内、抗菌药物使用前及抗菌药物有效治疗7 d 空腹上肢静脉血,测定患者体内 CD64、PCT 水平,同时行血常规、肝功能、肾功能、凝血功能检查。腹水非 SBP 组及对照组一次性采血测定以上指标。结果 SBP 组 CD64、血清 PCT 水平较腹水非 SBP 组和对照组明显升高（P ＜0．01）。 ROC 曲线分析结果显示 CD64和PCT 的敏感性和特异性分别为95．5％、93．8％和96．1％、85．2％。结论 CD64、PCT 可作为肝硬化 SBP 早期诊断、疗效判定的重要指标。     

The Expression of Calcitonin Gene-related Peptide on the Neurons Associated Zusanli (ST 36) in Rats

Objective:To investigate the biochemical characteristic of the neurons associated Zusanli(ST 36)in the rat by using Alexa Fluor 594 conjugated cholera toxin subunit B(AF594-CTB)neural tracing and calcitonin gene-related peptide(CGRP)fluorescent immunohistochemical techniques.Methods:Four male Sprague Dawley rats were injected with AF594-CTB into the corresponding area of the Zusanli in the human body.After 3 surviving days,the rat's spinal cord and dorsal root ganglia(DRGs)at lumbar segments were dissected following perfusion with 4%paraformaldehyde,cut into sections,and then stained with CGRPfluorescent immunohistochemical method.Results:AF594-CTB labeled sensory neurons were detected in the L3-L6 DRGs with high concentration in L4 DRG,and the labeled motor neurons located in the dorsolateral and intermediate regions of laminaⅨfrom L3-L5 segments with high concentration at L4.Meanwhile,CGRPpositive neural labeling distributed symmetrically on both sides of DRGs,anterior and dorsal homs of spinal cord.In the AF594-CTB labeled neurons,37%sensory neurons and 100%motor neurons expressed CGRPpositive.Conclusion:These findings present the morphological evidence to demonstrate that the sensory and motor neurons associated Zusanli in the rat distributed with segmental and regional patterns,and contained CGRP-expression.