BMP信号通路在华法林诱导的大鼠血管平滑肌细胞钙化中的作用

目的探讨骨形态发生蛋白(BMP)信号通路在华法林诱导的大鼠血管平滑肌细胞(VSMC)钙化中的作用。方法体外分离培养大鼠胸主动脉VSMC,将其随机分为正常对照组、高磷组、华法林(10μmol/L)干预组及华法林(10μmol/L)+维生素K(10μmol/L)干预组。对细胞进行钙含量和碱性磷酸酶(ALP)活性检测,茜素红染色检测钙结节,Western blot检测细胞中Runx2蛋白的表达变化,RT-PCR检测细胞中骨形态发生蛋白2(BMP-2)、Smad1及Runx2的表达变化。结果茜素红染色结果显示,与正常对照组和高磷组相比,华法林干预组钙化结节明显增多;钙含量测定结果与茜素红染色结果基本一致。与正常对照组和高磷组相比,华法林干预组ALP活性明显增加(P0.05),Runx2 mRNA和蛋白表达水平及BMP-2、Smad1 mRNA表达明显增高(P0.05);与华法林干预组相比,华法林+维生素K干预组细胞钙含量、ALP活性及BMP-2、Smad1、Runx2的表达均明显降低(P0.05)。结论BMP信号通路参与了华法林促进的大鼠VSMC钙化,其可能的作用机制是介导了VSMC的表型转化。     

氟骨症骨转换过程中转化生长因子-β超家族成员的表达

目的探讨氟中毒大鼠的骨转换调控过程,分析转化生长因子-β(transforming growth factor-β, TGF-β)超家族成员骨形态发生蛋白(BMP)在氟骨症骨转换过程中的作用。方法4周龄雄性Wistar大鼠随机分为对照组和氟化钠组,氟化钠组饮用含氟(F-)150 mg／L的水溶液,对照组饮用自来水。每4周处死一批动物,共计6次,采用RT-PCR方法、免疫组织化学方法以及Western blot等方法,分析氟中毒大鼠长骨组织TGF-β超家族成员BMP-2、BMP-7、TGF-β1 mRNA及蛋白的表达情况。结果氟中毒大鼠骨组织中BMP-2、BMP-7、TGF-β1mRNA表达持续高于对照组,蛋白主要表达在骺板软骨增生期和肥大期细胞、关节软骨细胞、成骨细胞以及韧带细胞等部位。结论氟中毒大鼠骨组织中TGF-β超家族成员通过调控成骨过程而参与了骨转换过程     

骨形成蛋白-7及其受体mRNA在大鼠肾间质纤维化模型中的表达

骨形成蛋白-7(bone morphogenetic pmtein-7，BMP-7)属于转化生长因子β(TGF-β)超家族，是一个重要的肾脏保护细胞因子，目前有关BMP-7及其受体在肾间质纤维化中的表达国内尚未见报道。本实验观察单侧输尿管梗阻(UUO)大鼠肾脏BMP-7及其受体的动态表达，为以后研究提供实验依据。     

内质网应激介导了高糖引起的血管平滑肌细胞钙化

目的探讨内质网应激是否介导了高糖引起的血管平滑肌细胞(VSMC)钙化。方法体外高糖(35μmol/L D-葡萄糖)处理VSMC模拟糖尿病环境,观察高糖是否引起VSMC内质网应激反应和凋亡,探索高糖是否引起VSMC表型转化(收缩型转变为成骨样细胞),观察内质网应激诱导剂和抑制剂对VSMC钙化的影响,碱性磷酸酶(ALP)活性、钙沉积和骨分化转录因子(Runx2和Osterix)通过比色法、O-cresolphthalein法和Western blot测定。结果应用35μmol/L D-葡萄糖分别处理VSMC不同时间,可以上调VSMC内质网应激标志蛋白表达、ALP活性、钙沉积和骨分化标志蛋白;然而,4-PBA预处理抑制VSMC内质网应激反应的同时,也能阻断高糖引起的VSMC钙化,表现为ALP活性、钙沉积和骨分化标志蛋白下降。结论高糖可以激活VSMC的内质网应激和凋亡,进而促VSMC钙化的发生,提示可能内质网应激介导了此激活过程。     

骨形态发生蛋白-4在乳腺癌侵袭及转移中作用的研究进展

骨形态发生蛋白4（BMP-4），又名BMP-2B，是转化生长因子-β（TGF—β）超家族成员之一，不但有促进人体胚胎发育及细胞增殖、分化、凋亡的能力，而且在肿瘤增殖、侵袭及转移中起有重要作用。该文结合最新研究进展，分析BMP4和BMP4受体在乳腺癌中的表达及其信号通路的影响因素和作用，为了解乳腺癌的发生发展，提供可能的分子机制。     

骨形态发生蛋白与血管钙化研究进展

近年血管钙化的细胞分子机制研究结果表明,血管钙化形成过程是一个与骨发育相似的,主动的,可预防和可逆转的高度可调控的生物学过程。1965年Urist发现骨形态发生蛋白（bonemorphogeneticproteins,BMPs）．到目前为止,已发现30余种成员,其中以BMP-2的作用最为重要。它们是一类唯一能够单独诱导骨组织形成的局部生长因子,     

过氧化体增殖物激活型受体γ抑制转化生长因子β1诱导的血管平滑肌细胞钙化

目的探讨过氧化体增殖物激活型受体γ(PPARγ)对转化生长因子β1(TGF-β1)诱导的大鼠主动脉血管平滑肌细胞(VSMC)钙化的作用。方法体外培养大鼠主动脉VSMC,先分为正常对照组、不同浓度TGF-β1组(1、2、4、8μg/L),观察TGF-β1对VSMC的影响;再分为正常对照组、钙化组(TGF-β1 4μg/L)、罗格列酮(RSG,20μmol/L)组、钙化+罗格列酮(RSG,20μmol/L)组,观察PPARγ激动剂罗格列酮对VSMC钙化后的作用,对细胞进行钙含量和碱性磷酸酶(ALP)活性检测,茜素红S染色检测钙化结节的形成情况,Western blot检测VSMC标志物α平滑肌肌动蛋白(α-SMA)、PPARγ、成骨样细胞标志物Runt相关转录因子2(Runx2)的蛋白表达情况。结果与正常对照组相比,TGF-β1处理后的VSMC钙盐沉积和ALP活性明显升高(P0.05),且TGF-β1浓度为4μg/L时作用最明显,成骨样细胞标志物Runx2表达明显升高(P0.05),同时平滑肌细胞标志物α-SMA表达减少(P0.05)。而加入罗格列酮后,VSMC的钙盐沉积和ALP活性明显降低(P0.05),α-SMA、PPARγ表达明显上升(P0.05),相反,Runx2的表达则明显受到抑制(P0.05)。结论 TGF-β1可以诱导VSMC向成骨样细胞分化和钙化,而PPARγ激动剂罗格列酮可以抑制TGF-β1诱导下VSMC钙化的发生。     

高磷对大鼠胸主动脉血管环中骨形成蛋白2表达的影响

目的:探讨骨形成蛋白-2(Bone morphogenetic protein 2,BMP-2)在高磷诱导的大鼠胸主动脉血管环钙化中的作用。方法:选取8~10周龄健康雄性SD大鼠,体外培养大鼠胸主动脉血管环,血管环按随机数字表法随机分为2组:正常对照组和高磷组。血管环培养7d和14d后,采用von Kossa染色及邻甲酚酞络合酮比色法检测大鼠胸主动脉血管环钙化情况;免疫组织化学方法检测血管环BMP-2的表达。体外采用组织块贴壁法培养原代大鼠胸主动脉平滑肌细胞,利用10mmol/Lβ-甘油磷酸制备血管平滑肌细胞钙化模型,细胞随机分为2组:正常对照组、高磷组(10mmol/Lβ-甘油磷酸)。采用茜素红染色及邻甲酚酞络合酮比色法检测细胞钙化情况,RT-PCR和Western blot方法检测细胞培养7d和14d的BMP-2mRNA及蛋白表达,并观察短时间内不同时间点BMP-2蛋白表达情况。结果:体外血管环培养7d、14d后,与正常对照组相比,高磷组钙含量明显增加(P0.05);与7d高磷组血管环钙含量相比,14d高磷组血管环钙含量明显增加(P0.05)。免疫组织化学结果显示,与正常对照组相比,高磷组BMP-2表达增加(P0.05);与7d高磷组相比,14d高磷组血管环钙含量明显增加(P0.05)。细胞培养7d、14d后,与正常对照组相比,高磷组钙盐沉积及钙含量明显增高(P0.05);RTPCR和Western Blot显示,与正常对照组相比,高磷组BMP-2mRNA及蛋白表达增加。进一步动态观察BMP-2蛋白的表达变化,结果显示正常对照组及高磷组BMP-2蛋白表达随时间延长呈增强趋势(P0.05)。结论:BMP-2可能参与了高磷诱导的血管钙化的发生发展。     

血清BMP-2、BMP-4检测在慢性肾脏病大鼠主动脉钙化中的意义

目的建立慢性肾脏病(CKD)大鼠主动脉钙化模型,探讨血清骨形态发生蛋白2(BMP-2)、骨形态发生蛋白4(BMP-4)与主动脉钙化程度的相关性。方法 55只SD大鼠随机分为对照组(20只)和CKD组(35只,由于预实验中CKD组大鼠后期存在死亡现象,为保证实验过程每个时间的实验样本量充足,因此CKD组样本量较对照组多),分别于第2、4、6、8周收集血液、尿液、主动脉、肾脏;肾脏HE染色评估肾脏病理改变;茜素红染色法、钙含量测定评估主动脉钙化程度; ELISA测定大鼠血清BMP-2、BMP-4水平;免疫组织化学检测主动脉BMP-2、BMP-4蛋白表达。结果与对照组各时间点相比,CKD组大鼠24 h尿蛋白定量、血清尿素氮、肌酐、胱抑素C水平明显增加(P0.05)。肾脏HE染色显示,自第2周起,肾小管出现扩张,小管内棕黄色物质沉积,随时间进展,逐渐出现间质纤维化,肾小球部分萎缩,血管减少,炎性细胞浸润。茜素红染色随时间进展,第8周出现大量橘红色颗粒物沉积伴钙结节形成。CKD组钙含量明显增加且与血清BMP-2 (r=0.929,P0.01)及BMP-4 (r=0.702,P0.01)明显正相关。同时,自模型成功建立第4周,主动脉BMP-2、BMP-4蛋白表达即明显增加。结论血清BMP-2、BMP-4表达增加可作为CKD血管钙化的血清标志物。     

骨碎补总黄酮对大鼠骨髓间充质干细胞成骨分化过程中TGF-β1、BMP-2分泌的影响

将大鼠分为空白对照组、经典组和骨碎补组,采用全骨髓贴壁法分离纯化大鼠骨髓间充质干细胞(BMSCs),检测BMSCs成骨分化过程碱性磷酸酶活性、矿化结节、转化生长因子-β1(TGF-β1)和骨形态发生蛋白2(BMP-2)的分泌.结果显示,骨碎补总黄酮促进成骨分化同时增加TGF-β1、BMP-2分泌(均P＜0.05).骨碎补总黄酮可能通过上调二者的表达促进成骨,对骨质疏松症可能有积极的治疗意义.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

N-acetyl-L-cysteine combined with mesalamine in the treatment of ulcerative colitis： Randomized,placebo-controlled pilot study

AIM： To evaluate the effectiveness and safety of oral N-acetyl-L-cysteine （NAC） co-administration with mesalamine in ulcerative colitis （UC） patients.
METHODS： Thirty seven patients with mild to moderate UC were randomized to receive a four-wk course of oral mesalamine （2.4 g/d） plus N-acetyl-L-cysteine （0.8 g/d） （group A） or mesalamine plus placebo （group B）. Patients were monitored using the Modified Truelove-Witts Severity Index （MTWSI）. The primary endpoint was clinical remission （MTWSI ≤ 2） at 4 wk. Secondary endpoints were clinical response （defined as a reduction from baseline in the MTWSI of ≥ 2 points） and drug safety. The serum TNF-α, interleukin-6, interleukin-8 and MCP-1 were evaluated at baseline and at 4 wk of treatment. RESULTS： Analysis per-protocol criteria showed clinical remission rates of 63% and 50% after 4 wk treatment with mesalamine plus N-acetyl-L-cysteine （group A） and mesalamine plus placebo （group B） respectively （OR = 1.71; 95% CI： 0.46 to 6.36; P = 0.19; NNT = 7.7）. Analysis of variance （ANOVA） of data indicated a significant reduction of MTWSI in group A （P = 0.046） with respect to basal condition without significant changes in the group B （P = 0.735） during treatment. Clinical responses were 66% （group A） vs 44% （group B） after 4 wk of treatment （OR = 2.5; 95% CI： 0.64 to 9.65; P = 0.11; NNT = 4.5）. Clinical improvement in group A correlated with a decrease of IL-8 and MCP-1. Rates of adverse events did not differ significantly between both groups.
CONCLUSION： In group A （oral NAC combined with mesalamine） contrarily to group B （mesalamine alone）, the clinical improvement correlates with a decrease of chemokines such as MCP-1 and IL-8. NAC addition not produced any side effects.     

Delorme's transrectal excision for internal rectal prolapse

Surgical therapy of functional outlet obstruction in patients with internal rectal intussusception may include abdominal, perineal, or transrectal procedures. Because abdominal procedures often result in significant physiologic impact but unrelieved constipation, the authors have elected Delorme's transrectal excision for management of these patients. Since a short-term placebo effect attends many therapies, this report describes results of transrectal excision only after a threeyear postoperative period. Delorme's transrectal excision of internal intussusception accomplished sustained symptomatic relief in over 70 percent of otherwise refractory constipated patients. The association of internal intussusception with other abnormalities underscores the importance of defining both anatomic and functional components when selecting patients whose constipation may require surgical therapy. Critical technical elements, surgical pitfalls, and potential complications of the procedure are discussed.Poster presentation at the meeting of The American Society of Colon and Rectal Surgeons, Toronto, Canada, June 11 to 16, 1989.     

The oral glucose tolerance test: A comparison of the time points on the basis of limit values,normal dispersion,and reproducibility

Summary Time points in the glucose tolerance test (GTT) are compared on the basis of limit values, dispersion within a reference population, and reproducibility. We suggest using the distance between a limit value and the median reference value as a measure of the magnitude of abnormality. The distance between 140 mg/100 ml and the median fasting plasma glucose value is chosen as a standard distance and limits for other points in the GTT are calculated to equal this standard distance of abnormality. We suggest that the probability of correctly interpreting an inividual result is directly related to the reproducibility of the test and inversely related to the percentage of the total range of values which is dispersed among the normal population. The ratio of reproducibility to percentage normal dispersion is proposed as an index of the probability of correctly interpreting an individual result. According to this index, the probability of correct interpretation varies in order: fasting plasma glucose concentration>3-h>2-h>0.5-h>1-h plasma glucose concentration.