Skeletal unloading induces resistance to insulin-like growth factor-I (IGF-I) by inhibiting activation of the IGF-I signaling pathways.

We showed that unloading markedly diminished the effects of IGF-I to activate its signaling pathways, and the disintegrin echistatin showed a similar block in osteoprogenitor cells. Furthermore, unloading decreased alphaVbeta3 integrin expression. These results show that skeletal unloading induces resistance to IGF-I by inhibiting activation of the IGF-I signaling pathways at least in part through downregulation of integrin signaling. INTRODUCTION: We have previously reported that skeletal unloading induces resistance to insulin-like growth factor-I (IGF-I) with respect to bone formation. However, the underlying mechanism remains unclear. The aim of this study was to clarify how skeletal unloading induces resistance to the effects of IGF-I administration in vivo and in vitro with respect to bone formation. MATERIALS AND METHODS: We first determined the response of bone to IGF-I administration in vivo during skeletal unloading. We then evaluated the response of osteoprogenitor cells isolated from unloaded bones to IGF-I treatment in vitro with respect to activation of the IGF-I signaling pathways. Finally we examined the potential role of integrins in mediating the responsiveness of osteoprogenitor cells to IGF-I. RESULTS: IGF-I administration in vivo significantly increased proliferation of osteoblasts. Unloading markedly decreased proliferation and blocked the ability of IGF-I to increase proliferation. On a cellular level, IGF-I treatment in vitro stimulated the activation of its receptor, Ras, ERK1/2 (p44/42 MAPK), and Akt in cultured osteoprogenitor cells from normally loaded bones, but these effects were markedly diminished in cells from unloaded bones. These results were not caused by altered phosphatase activity or changes in receptor binding to IGF-I. Inhibition of the Ras/MAPK pathway was more impacted by unloading than that of Akt. The disintegrin echistatin (an antagonist of the alphaVbeta3 integrin) blocked the ability of IGF-I to stimulate its receptor phosphorylation and osteoblast proliferation, similar to that seen in cells from unloaded bone. Furthermore, unloading significantly decreased the mRNA levels both of alphaV and beta3 integrin subunits in osteoprogenitor cells. CONCLUSION: These results indicate that skeletal unloading induces resistance to IGF-I by inhibiting the activation of IGF-I signaling pathways, at least in part, through downregulation of integrin signaling, resulting in decreased proliferation of osteoblasts and their precursors.     

Experimental Study on Allogenic Decalcified Bone Matrix as Carrier for Bone Tissue Engineering

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rab-bits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as con-trol. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts,trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the sur-face of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good bio-compatibility and ectopic induced osteogentic activty.     

碘杂环化合物对离体豚鼠心乳头肌动作电位和收缩力的作用

采用细胞内微电极记录技术,同步观察了3,6-[二甲氨基]-二苯并碘因甲酸盐(IHC-64)对豚鼠心乳头肌细胞动作电位和收缩力的作用。50μmol/L IHC-64抑制心肌收缩力,而不影响快反应动作电位。增加IHC-64浓度,动作电位0相最大峰值(APA)、除极速率(dp/dt_(max))和复极50％和90％时程(APD_(50)、APD_(90))被明显抑制。IHC-64抑制慢反应动作电位,提高细胞外钙浓度可拮抗这种抑制。结果提示,IHC-64主要抑制慢Ca~(2+)内流,同时也部分抑制快Na~+内流,它可能是一种新型B类钙通道阻滞剂。     

Differential antagonism of the behavioral depressant and hypothermic effects of 5'-(N-ethylcarboxamide) adenosine by theobromine

The methylxanthine, theobromine (3,7-dimethylxanthine), was tested in mice, to determine whether theobromine could function in vivo as an adenosine receptor antagonist, in keeping with its reported in vitro effects as a blocker of agonist binding to the adenosine A-1 receptor. Theobromine doses, which themselves had no direct effects on spontaneous locomotor activity, completely blocked N6-cyclohexyladenosine-induced suppression of locomotor activity but were without effect on 5'-N-ethylcarboxamide adenosine (NECA)-induced decreases in motor activity. In contrast to the specific antagonism, theobromine blocked the hypothermia induced by both of these adenosine analogs. These results demonstrate that theobromine is an active in vivo adenosine receptor antagonist and that the antagonism of N6-cyclohexyladenosine sensitive systems occurs even though theobromine does not stimulate spontaneous locomotor activity. Thus, the behavioral stimulant effects of methylxanthines may be more related to effects on NECA-sensitive systems, which are not blocked by theobromine. The use of in vivo differences in the effects xanthine may provide a useful tool in the development of compounds to probe the mechanisms of caffeine induced CNS effects.     

潘生丁增强氨甲蝶呤的抗肿瘤作用

外源性核苷能抵消抗代谢药对肿瘤细胞的杀伤作用;核苷转运抑制剂潘生丁则能阻断核苷的这种抵消作用,从而增强抗代谢药的细胞毒性。本研究证明,胸苷和次黄嘌呤可明显抵消氨甲蝶呤对L1210细胞的杀伤作用,潘生丁则能有效地阻断核苷的抵消作用;潘生丁和两性霉素B合用可明显增强氨甲蝶呤对小鼠S180肉瘤的抑制作用,但不增强氨甲蝶呤对动物的毒性。提示潘生丁有可能应用于肿瘤联合化疗。     

尿流改道及膀胱重建术的现状与展望

膀胱全切术后尿液的贮存与排出一直是未能满意解决的问题。自从1852年Simon报道输尿管乙状结肠吻合以来，经过一个多世纪的不断改进与创新，特别是1982年Kock用去管重建法制作贮尿囊的可控膀胱以来，尿流改道与膀胱重建术有了跨时代的进步和发展，显地提高了患术后生活质量。     

The correlation between TNF-α, IL-6 in cervical mucous during follicular development and ovulation stimulation in different protocols

Objective To investigate the correlation between TNF-α and IL-6 levels in cervical mucous during follicular development and ovulation stimulation in different protocols.Methods 36 infertile women were set up as experimental groups,divided into CC, HMG, IVF-ET group,each group consisted of 12 infertile women and 15 women with normal menstrual cycles were choiced as control group.Cervical mucous during follicular phase, luteal phase and ovulation phase were collected.TNF-α, IL-6 levels in cervical mucous were measured by radioimmunology assay (RIA).Follicular development were monitored by transvaginal ultrasonagraphy.Results (1) TNF-α levels in cervical mucous of experimental groups and control group were periodically various among the reproductive cycle.It increased during follicular phase, reached to peak during ovulation phase, and decreased during luteal phase (P＜0.05).IL-6 levels had no obvious periodical changes.(2) Compared with CC and control group, levels of TNF-α,IL-6 in HMG and IVF-ET group were significantly higher (P＜0.05).(3) Levels of TNF-α and IL-6 in cervical mucous were positively correlated with the dominant follicle diameter (r=0.261, r=0.192 respectively,P＜0.05).(4) TNF-α and IL-6 showed positive correlation in the reproductive cycle (r=0.782,P＜0.05).Conclusions (1) TNF-α level shows a cyclic change in the reproductive cycle and peaks during ovulation,whereas IL-6 level does not.(2) TNF-α and IL-6 may play a certain role in the process of follicular development and ovulation.(3)The levels of TNF-α and IL-6 are up-regulated by gonadotrophic hormone.(4) TNF-α and IL-6 may have coordination properties and participate in the same biological effects.     

大白鼠皮肤Ⅱ度、Ⅲ度碱烧伤模型复制

目的 根据大白鼠皮肤碱烧伤创面早期病理组织学观察复制Ⅱ、Ⅲ度碱烧伤模型,研究碱烧伤的临床治疗.方法 Wistar纯种健康大白鼠20只,以不同浓度2mol/l、2.5mol/l、5mol/lNaOH,作用时间分别为30秒、45秒、60秒、75秒,涂抹于脱毛后的表皮.结果 2mol/L(60s)、2.5mol/L(45s),5mol/L(30s)即可直接造成Ⅱ度～深Ⅱ度烧伤;2mol/L(75s)、2.5mol/L(60s)、5 mol/L(45s)即可造成Ⅲ度烧伤,其烧伤深度与NaOH溶液浓度和作用时间呈正比.结论 Ⅱ度、Ⅲ度碱烧伤均为渐进性烧伤,与文献中介绍的潜拙样损伤似乎不同.另外,不同浓度NaOH溶液在相同时间对皮肤组织的损伤及同一浓度NaOH溶液在不同时间对大白鼠皮肤的损伤病理学变化均有差异.     

中药外治法改善类风湿性关节炎患者关节功能的作用

目的:观察自拟中药汽疗薰蒸方治疗类风湿性关节炎的临床疗效,以及对患者免疫功能的调节。方法:共选入150例类风湿性关节炎患者,为2002-01/2005-01哈尔滨医科大学附属第一医院中西医结合科住院患者,随机分为治疗组100例和对照组50例。治疗组患者进行中药汽疗(方药组成:制川乌15kg,制草乌15kg,乌梢蛇1条,全蝎5g,蜈蚣2条,威灵仙20g,海风藤20g,伸筋草20g,透骨草20g,羌活15g,独活15g,防已15g,地龙15g,鸡血藤25g),1次/d,30min/次;对照组患者给予常规抗类风湿性关节炎药物治疗:青霉素静脉滴注,1次/d,同时口服西乐葆片,200mg/次,2次/d,治疗1个疗程(10d)后,观察两组患者的临床症状中关节肿胀、疼痛、压痛、晨僵和功能障碍的改善情况,并对主要指标血沉、类风湿因子、C反应蛋白、免疫球蛋白(IgA,IgG)等进行评估和监测。结果:参选的150人全部完成数据采集,无脱落者。①两组疗效比较:治疗组总有效率优于对照组(94%,66%,P<0.01)。②治疗后症状体征各运动功能评分:治疗组关节疼痛度、关节压痛度、活动度、肿胀度、晨僵时间均低于对照组[(0.92±0.56),(1.19±0.60)分;(3.13±1.89),(4.21±2.42)分;(0.54±0.43),(0.98±1.44)分;(4.46±2.80),(8.05±3.28)分;(18.52±10.23),(24.78±11.21)min,P<0.01]。③治疗后实验室指标变化:治疗组血沉、类风湿因子、C反应蛋白、IgA、IgG均低于对照组[(25.15±14.28),(46.23±14.59)mm/h;(29.43±15.98),(46.44±19.67)滴度;(14.57±8.90),(19.43±10.12)mg/L;(2.16±1.40),(2.98±1.95)g/L;(14.82±4.37),(18.94±5.51)g/L,P<0.01]。结论:中药汽疗治疗类风湿性关节炎疗效显著,可明显改善人体的免疫功能,且方法简便易行。