胰岛素样生长因子-1对成骨细胞生长影响的实验研究

目的:探讨胰岛素样生长因子-1对成骨细胞生长增殖的影响.方法:采用酶消化法获取兔骨膜成骨细胞,取第三代成骨细胞与0.1ng/ml、1ng/ml、10ng/ml的胰岛素样生长因子-1共同体外培养,在第1～7d观察细胞的形态、生长特点,MTT法测定细胞增殖情况,并通过细胞计数绘制细胞生长曲线.结果:不同浓度的胰岛素样生长因子-1对成骨细胞的生长增殖与对照组比较均有显著性差异(P＜0.01),各浓度之间对成骨细胞的生长增殖亦存在显著性差异(P＜0.05).结论:胰岛素样生长因子-1对成骨细胞生长增殖具有促进作用,在0.1～10ng/ml范围内此种作用与浓度呈正相关.     

胰岛素样生长因子-1对成骨细胞生长影响的实验研究

目的探讨胰岛素样生长因子-1对成骨细胞生长增殖的影响.方法采用酶消化法获取兔骨膜成骨细胞,取第三代成骨细胞与0.1ng/ml、1ng/ml、10ng/ml的胰岛素样生长因子-1共同体外培养,在第1～7d观察细胞的形态、生长特点,MTT法测定细胞增殖情况,并通过细胞计数绘制细胞生长曲线.结果不同浓度的胰岛素样生长因子-1对成骨细胞的生长增殖与对照组比较均有显著性差异(P＜0.01),各浓度之间对成骨细胞的生长增殖亦存在显著性差异(P＜0.05).结论胰岛素样生长因子-1对成骨细胞生长增殖具有促进作用,在0.1～10ng/ml范围内此种作用与浓度呈正相关.     

强骨宝方对体外培养成骨细胞影响的实验研究

目的:探讨不同浓度强骨宝方提取液直接添加对体外培养成骨细胞增殖、分化与矿化功能的影响。方法:采用胰蛋白酶-Ⅱ型胶原酶消化法从2只1~2日龄SD乳鼠颅盖骨中分离出成骨细胞,鉴定细胞并传代培养后,分为对照组与4个实验组,实验组分别用终浓度为100、50、10、5μg/ml的强骨宝方提取液加入成骨细胞培养体系,对照组用不含强骨宝方提取液的培养基培养,应用MTT比色法、ALP含量测定、矿化结节形成等分别观察其对成骨细胞增殖、分化及矿化能力的影响。结果:100、50及10μg/ml浓度的强骨宝方提取液均具有促进体外成骨细胞增殖、分化与矿化的作用,以100μg/ml与50μg/ml促进作用最强。结论:每毫升含生药2g的强骨宝方提取液,pH=7.0,50μg/ml的添加浓度可能最适合于体外培养成骨细胞的增殖、分化及矿化成骨能力。     

酸性成纤维细胞生长因子及表皮生长因子对兔韧带细胞增殖的影响

目的：观察酸性成纤维细胞生长因子(aFGF)和表皮生长因子(EGF)对内侧副韧带(MCL)和前十字韧带(ACL)细胞增殖行为的影响。方法：培养10周龄新西兰白兔内侧副韧带和前十字韧带细胞，在培养液中分别加入aFGF和EGF，以XTT方法测定细胞的增殖行为。结果：aFGF在1ng／ml时即对两种细胞具有显著的促进增殖作用，其浓度达50ng／ml时，对MCL细胞的促进作用最大，达100ng／ml时对ACL细胞的促进作用最大。EGF在0．78ng／ml时即对MCL细胞有显著的促增殖作用，在1．56ng／ml时始对ACL细胞有显著的促增殖作用，其浓度达3．125ng／ml时对2种细胞的促进作用最大。aFGF和EGF在超过其最佳浓度后，随浓度升高促进作用均下降。结论：aFGF和EGF可以促进韧带成纤维细胞增殖。     

rhIGF-1对成骨细胞增殖、BMP-2及Cbfa1基因表达的影响

目的通过重组人胰岛素样生长因子(rhIGF-1)对体外成骨细胞增殖、骨形态蛋白-2(BMP-2)及核心结合因子1(Cbfa1)基因表达的影响,探讨rhIGF-1对骨代谢影响的可能机制。方法不同浓度的rhIGF-1(0、10、50、100ng/ml)刺激体外培养的大鼠成骨细胞,采用噻唑蓝(MTT)法检测细胞增殖能力,用半定量RT-PCR法检测成骨细胞BMP-2、Cbfa1基因的表达。结果rhIGF-1可明显促进成骨细胞增殖(P0.05),并促进成骨细胞BMP-2、Cbfa1基因的表达(P0.01),具有浓度依赖性。结论rhIGF-1可促进成骨细胞的增殖、分化及成熟,可能是通过增强BMP-2、Cbfa1基因的表达来实现的。     

rhIGF-1对成骨细胞增殖、分化及骨保护素、骨保护素配体基因mRNA表达的影响

目的 观察不同剂量rhIGF-1对成骨细胞增殖，分化及骨保护素，骨保护素配体mRNA基因表达的影响，为明确骨保护素，骨保护素配体在骨质疏松症发病的作用机理及rhIGF-1在临床中应用提供实验依据。方法 取2代培养的大鼠成骨细胞，在rhIGF-1为0ng/ml,10ng/ml,20ng/ml,50ng/ml的浓度中培养。观察细胞的生长及钙结节的形成，MTT法，碱性磷酸酶（ALP），骨钙素（OCN）测定细胞增殖和分化，RT－PCR测定rhIGF-1对成骨细胞骨保护素，骨保护素配体基因mRNA表达的影响。结果 成骨细胞在7d可铺满瓶壁，30d可形成钙结节，rhIGF-1可促进成骨细胞的增殖，ALP和OGN的分泌，促进骨保护素，骨保护素配体基因的表达，以促进骨保护素表达明显，在rhIGF-1为10ng/ml时作用明显。结论 rhIGF-1可促进大鼠成骨细胞的增殖，分化，及骨保护素，骨保护素配体基因mRNA的表达，骨保护素mRNA表达显。rhIGF-1可能通过影响骨保护素，骨保护素配体而调节成骨细胞破骨细胞的平衡，使骨重建，从而防治骨质疏松症。     

TNF-α对体外培养的人牙囊细胞增殖特性及碱性磷酸酶的影响

目的:检测不同浓度TNF-对体外培养人牙囊细胞增殖活性和碱性磷酸酶活性的影响。方法:第5代人牙囊细胞接种于96孔培养板上,分别与不同浓度的TNF-共同孵育,检测TNF-对人牙囊细胞的增殖活性和碱性磷酸酶活性的影响。结果:TNF-在浓度为10～50ng/ml,时间为3天时促进人牙囊细胞的增殖,浓度为10～200ng/ml时抑制碱性磷酸酶活性。结论:TNF-在特定的浓度和时间促进人牙囊细胞的增殖,抑制牙囊细胞的成骨特性。     

基因重组人生长激素对体外培养的肝癌细胞增殖的影响及配体调控

目的 了解基因重组人生长激素(rhGH)对体外培养的Bel-7402细胞增殖的影响,并探讨rhGH对肝癌细胞生长激素受体(GHR)的调控作用.方法 分别采用肿瘤细胞计数、MTT比色法和集落形成实验等方法 了解Bel-7402细胞株在不同浓度rhGH(0、1、10、100、1000、10 000 ng/ml)作用下的药物敏感性,计算细胞生长率;用3H-TdR掺入法研究肿瘤细胞DNA代谢情况;应用放射配体法了解Bel-7402细胞GHR的表达,以及rhGH在上述浓度下对肝癌细胞GHR的影响.结果 rhGH对体外培养的Bel-7402细胞的生长有一定程度促进作用,表现为rhGH在100 ng/ml浓度下促进肝癌细胞增殖较为显著,rhGH在其他浓度(1、10、1000、10000 ng/ml)的部分时段,对肝癌细胞的增殖虽亦有一定程度的促进作用,但总体效果较rhGH在100 ng/ml浓度时弱;放射配体法发现Bel-7402细胞表达GHR,在rhGH作用后24 h,Bel-7402细胞GHR的位点数量(103个/细胞)在10、100 ng/ml组较对照组显著增加,在10000ng/ml组较对照组显著减少.结论 一定浓度范围rhGH对Bel-7402细胞的增殖有促进作用,原因可能与Bel-7402细胞表达GHR有关;同时rhGH对Bel-7402细胞的GHR有一定调控作用.     

骨组织工程中促细胞粘附因子bFGF的研究

目的 针对骨组织工程中成骨细胞与基质材料间促粘附这一重要问题,研究碱性成纤维细胞生长因子(bFGF)对成骨细胞粘附特性的影响。方法 取兔骨髓基质干细胞来源的成骨细胞体外培养,分别用Ong/ml、10ng/ml、100ng/ml浓度的bFGF诱导培养,观察接种后各时间点成骨细胞粘附情况,体视学计量粘附细胞数量。结果 10ng/ml bFGF诱导成骨细胞24小时后,细胞粘附数量增加最明显,显著高于对照组及100ng/ml组(P<0.01)。实验还发现,成骨细胞粘附最快发生在接种后4小时内。结论 一定浓度碱性成纤维细胞生长因子具有促进成骨细胞粘附特性的作用。     

骨组织工程中促细胞粘附因子bFGF的研究

目的：针对骨组织工程中成骨细胞与基质材料间促粘附这一重要问题,研究碱性成纤维细胞生长因子（bFGF)对成骨细胞粘附特性的影响。方法：取兔骨髓基质干细胞来源的成骨细胞体外培养,分别用0ng/ml、10ng/ml、100ng/ml浓度的bFGF诱导培养,观察接种后各时间点成骨细胞粘附情况,体视学计量粘附细胞数量。结果：10ng/mlbFGF诱导成骨细胞24小时后,细胞粘附数量增加最明显,显高于对照组及100ng/ml组（P＜0．01）。实验还发现,成骨细胞粘附最快发生在接种后4小时内。结论：一定浓度碱性成纤维细胞生长因子具有促进成骨细胞粘附特性的作用。     

生长激素在体外对人胃癌细胞的作用

目的　研究重组人生长激素 (rhGH)在体外对人胃癌细胞生长的影响。方法　实验分组 :对照组、rhGH组、奥沙利铂 (L OHP)组和rhGH L OHP组 ,利用体外细胞培养、MTT比色技术及流式细胞仪等方法 ,测定不同浓度的rhGH对人胃癌细胞株BGC82 3生长曲线、细胞抑制率、细胞周期和增殖指数 (PI)的影响。结果　rhGH在体外无明显促进BGC82 3细胞分裂增殖 ,rhGH组与对照组、rhGH L OHP组与L OHP组比较差异均无显著性 (P >0 .0 5 ) ,且生长曲线没有升高 ;rhGH L OHP组与对照组比较或rhGH L OHP组与对应的rhGH组配对比较 ,细胞抑制率增加 ,阻滞于G0 ～G1期的细胞数增加 ,S期细胞明显减少 ,PI明显降低 (P <0 .0 1)。rhGH L OHP组与L OHP组比较 ,细胞抑制率呈现升高趋势 ,而PI呈下降趋势 ,提示rhGH与抗肿瘤药合用可以增强抗肿瘤药对肿瘤细胞的杀伤作用。结论　rhGH在体外无明显促进胃癌细胞的分裂增殖 ,与抗肿瘤药合用可提高抗肿瘤药的疗效。     

rhbFGF和rhEGF对成纤维细胞的促增殖作用

目的：掌握人皮肤成纤维细胞的培养技术，了解该细胞的生长特性；研究rhbFGF和rhEGF在不同浓度下对人皮肤成纤维细胞增殖的影响，为临床准确定量应用这两种因子修复创面、加速创面愈合提供理论依据。方法：体外培养人皮肤成纤维细胞，用不同浓度的rhbFGF和rhEGF干预细胞，四甲基偶氮唑盐比色法（MTT）检测细胞的活性，选择最适浓度生长因子干预的细胞，用流式细胞仪检测细胞的DNA倍体情况。结果：不同浓度的rhbFGF和rhEGF分别对成纤维细胞干预48h后，显示rhbFGF在50μg／L浓度时MTT值最大，rhEGF在80μg／L浓度时MTT值最大；并检测出在最适浓度下细胞倍体的含量。结论：应用最佳浓度的rhbFGF和rhEGF可以更快的促进创面愈合，减少瘢痕的形成。     

Postnatal human lung growth.

Standard morphometric methods were applied to the lungs of 36 boys and 20 girls aged from 6 weeks to 14 years, dying as a result of trauma or after short illnesses. Individual lung units, alveolar dimensions, and number of alveoli per unit area and volume did not differ between boys and girls, but boys had bigger lungs than girls for the same stature. This resulted in a larger total number of alveoli and a larger aveolar surface area in boys than in girls for a given age and stature. There may be more respiratory bronchioles in boys than girls. There was rapid alveolar multiplication during the first two years of life and alveolar dimensions and number of alveoli per unit area and volume did not change much during this period. There was little or no increase in the total number of alveoli after the age of 2 years but the data are hard to interpret. There is a wide scatter of the total number of alveoli in the growing lung, in keeping with the observation that the total number of alveoli is very variable in adults. Prediction data are given for the various morphometric variables studied.     

基因重组人生长激素对人肝癌细胞系生长的影响

目的 研究基因重组人生长激素(rhGH)作用人肝癌细胞系后细胞周期动力学的改变，了解其对人肝癌细胞系生长的影响。方法 不同浓度rhGH(0、2、5、25、50、150、500μg／L)作用于人肝癌细胞系HepG2，采用流式细胞仪检测作用24、48、72h后的细胞周期动力学，分析其增殖指数、S期细胞百分比及细胞凋亡率的变化。结果 rhGH作用后的HepG2细胞周期动力学改变与作用时间无显著相关；与对照组比较，在生理浓度(25μg/L)rhGH作用下，PI及S％显著上升(P＜0．05)，但在超生理浓度(500μg／L)rhGH作用下，PI及S％无显著改变(P＞0．05)；另2、5、25、50、150、500μg/L rhGH均诱导人肝癌细胞凋亡，且呈浓度—效应关系。结论 在体外培养体系中，rhGH在低于或等于生理浓度时，促进人肝癌细胞增殖；在高于生理浓度时，抑制人肝癌细胞增殖。     

Role of transforming growth factor-alpha in human prostate cancer cell growth

Because of the influence of transforming growth factor-alpha (TGF alpha) on the cell growth in other cancer cell systems, we investigated the growth-regulatory role of TGF alpha in human prostate cancer cells. TGF alpha (5 ng/ml) stimulated LNCaP cell growth in monolayer to 60% of the level seen with dihydrotestosterone (DHT). Both DHT and TGF alpha increased cloning in soft agar twofold above that in controls. Metabolism of thymidine and uridine was also increased as evidenced by increased uptake of these macromolecule precursors. In addition, intracellular signalling as indicated by phosphatidyl inositol turnover was also increased by TGF alpha and DHT. Conditioned media contained TGF alpha by radioimmunoassay (RIA), transforming activity by rat kidney fibroblast (NRK) colony formation, and epidermal growth factor (EGF) receptor competable activity by radioreceptor assay. EGF receptors were present by binding assay and immunoprecipitation. These data demonstrate the presence of an autostimulatory growth loop in hormone-responsive human prostate cancer cells.     

Metabolic effects of recombinant human insulin-like growth factor-I in humans: comparison with recombinant human growth hormone

Many of the metabolic actions of growth hormone (GH( are mediated through insulin-like growth factors or somatomedins. Recombinant human insulin-like growth factor-I (rhIGF-I) has a dichotomous insulin-like and GH-like action when used in different clinical situations in humans. Its effets on carbohydrate metabolism show a prominent increase in total insulin sensitivity, causing hypoglycemia in higher doses and maintaining normal glucose homeostasis in lower doses. This polypeptide selectively stimulates whole body protein synthesis with no effect on proteolysis when given in doses of 100 g/kg subcutaneously twice daily for at least 5–7 days, effects which are indistinguishable from those of GH. This contrasts with the marked suppression of proteolysis observed when higher doses are given, similar to the effects observed with insulin. When used in combination with rhGH, rhIGF-I has a synergistic effect, improving total nitrogen retention in calorically deprived subjects, yet it does not cause any greater enhancement of whole body protein anabolism in normally fed volunteers than giving rhGH and rhIGF-I individually. This suggests a common pathway for IGF-I and GH enhancing protein anabolism in the normally fed state. rhIGF-I also stimulates linear growth in children with defects in the GH receptor. Recent data show that this potent growth factor has a potential advantage over GH in the treatment of severe protein catabolic states, particularly the glucocorticosteroid-dependent model, as it ameliorates the marked increase in protein catabolism caused by the steroids, but without a diabetogenic effect. Here, a brief overview is provided of available human data on the actions of this peptide on carbohydrate, lipid, and protein metabolism, linear growth, and its anabolic effects. rhIGF-I offers promise in the treatment of selective growth disorders and in protein catabolic and insulin-resistant states.     

Bombesin stimulates growth of human gastrinoma.

BACKGROUND. We have previously reported the first establishment and characterization of a functioning human gastrinoma (PT) xenograft. Bombesin, the equivalent of the mammalian gastrin-releasing peptide, has trophic effects on normal and neoplastic tissues of the gastrointestinal tract; the effects of gut hormones on the growth of gastrinoma are not known. The purpose of this study was twofold: (1) to determine the presence of various gut peptides in PT and (2) to determine the effect of bombesin on the growth of PT xenografts. METHODS. PT tumors were examined for expression (mRNA and protein) of various gut peptides by Northern hybridization and immunohistochemistry. In addition, PT xenografts were implanted as 3 mm2 pieces bilaterally subcutaneously in athymic nude mice. Mice were divided into two groups to receive either bombesin (5 micrograms/kg) or saline administered as intraperitoneal injections every 8 hours. Tumor area was measured twice weekly until mice were sacrificed (day 28), when tumor and normal pancreas were removed, weighed, and assayed for DNA and protein content. RESULTS. Both mRNAs and peptides of gastrin and chromogranin A were present in PT tumors. Bombesin significantly stimulated growth of PT tumors from day 18 until mice were sacrificed (day 28). As expected, bombesin stimulated pancreatic growth. CONCLUSIONS. We have demonstrated for the first time that bombesin is a trophic hormone for gastrinoma. The unique cell line PT contains gastrin and chromogranin A and will be a useful model to define the biologic mechanisms controlling the growth of human gastrinomas.     

Characteristic pattern of human prostatic growth with age

Aim: To study the characteristic pattern of the age-related growth of the human prostate gland. Methods: The volume (weight) of the prostate in 1,601 males, aged from newborn to 92 years, was determined by Bultrasonography. Results: Prostatic volume determination by B-ultrasonography in 1601 males (1301 normal subjects and 300 BPH patients) pointed out that the age-stratified growth of human prostate could be categorized into 4 life stages: (1) the first slow growing phase (from newborn to 9 years): the prostate grows slowly at a rate of 0.14g per year; (2) the first rapid growing phase (from 10 to 30 years): the prostate grows at a rate of 0.84 g per year; (3) the second slow growing phase (from 30 to 50 years), the prostate grows at a rate of 0.21 g per year; (4) the second rapid growing phase (from 50 to 90 years): the prostate grows at one of the following rates: in one group the growth rate is of 0.50 g per year and in the other 1.20 g per year, leading to benign prostatic hyperplasia (BPH). Conclu