2型糖尿病大鼠骨髓腔内PPARγ和Cbfα1 mRNA的表达与骨折愈合障碍的关系

目的:探讨2型糖尿病大鼠骨髓腔内过氧化物酶体增殖物激活受体γ (peroxisome proliferator-activated receptor gamma, PPARγ)和核心结合因子α1(core binding factor alpha 1,Cbfα1)表达的变化与2型糖尿病骨折愈合障碍之间的关系.方法:雄性SD大鼠随机分为对照组(n=15)和实验组(n=15)分别给予普通饲料、高糖高脂饲料喂养.8周后摘眼球取血,两组分别腹腔注射枸橼酸盐缓冲液和链脲佐菌素(STZ,35 mg/kg).2周后再次摘眼球取血,并建立大鼠左胫骨牵引成骨模型,牵引14 d后处死动物,收集血液及左胫骨,无菌分离双侧股骨.X线摄片比较两组胫骨牵引间隙内骨痂的形成情况.组织学观察牵引骨痂内微骨柱(MCF)与初始基质前沿(PMF)及骨折两端髓腔内脂肪细胞的形成情况,并计算骨折两端髓腔内脂肪细胞与骨髓腔面积百分比.RT-PCR法检测两组动物股骨骨髓腔内PPARγ和Cbfα1 mRNA的表达.结果:8周后实验组大鼠血清甘油三酯较对照组明显升高(P<0.01),血清总胆固醇较正常组升高(P<0.05),空腹血糖、血胰岛素水平无明显差异(P>0.05).10周后实验组大鼠空腹血糖、血清甘油三酯较对照组明显升高(P<0.01),血清总胆固醇较正常组升高(P<0.05),血胰岛素水平无明显差异(P>0.05),2型糖尿病大鼠模型建立.与对照组比较,2型糖尿病组大鼠X线摄片显示骨折断端之间牵引骨痂形成明显减少,骨痂组织学检查表现为MCF排列紊乱,PMF浅染;骨折两端髓腔内脂肪细胞及脂肪细胞占骨髓腔面积百分比明显增加(P<0.01);RT-PCR法检测显示双侧股骨骨髓腔内PPARγ mRNA表达上调(P<0.05), 而Cbfα1 mRNA的表达下调(P<0.05),而对照组则相反.2型糖尿病对早期骨折愈合表现为明显的抑制作用.结论:2型糖尿病大鼠骨折愈合障碍可能与2型糖尿病条件下骨髓腔内PPARγ mRNA表达增加而Cbfα1 mRNA的表达受抑制有关.     

神经生长因子促进大鼠胫骨骨折愈合的实验研究

目的 观察神经生长因子对大鼠胫骨骨折愈合以及胰岛素样生长因子表达的影响.方法 建立标准大鼠胫骨骨折模型,分别给予生理盐水和神经生长因子,术后1、2、4周分别进行X线骨痂评定、胫骨湿重称量以及半定量RT-PCR检测胰岛素样生长因子-1 mRNA的表达.结果 术后4周时,神经生长因子组骨痂量(132.63±5.98)mm3和胫骨湿重(0.98±0.12)g,对照组骨痂量(85.22±6.23)mm3和胫骨湿重(0.84±0.04)g,NGF组显著多于对照组(P<0.05);神经生长因子组的胰岛素样生长因子-1 mRNA的表达术后1周时为(0.82±0.17),术后2周时为(0.54±0.08),对照组分别为(0.55±0.20)、(0.41±0.09).NGF组显著高于对照组(P<0.05).结论 应用神经生长因子可以促进大鼠胫骨各组愈合,同时促进骨折愈合过程中IGF-1表达.     

神经生长因子对失神经状态下大鼠骨折愈合的影响?

目的:探讨神经生长因子(NGF)对脊髓完全横断伴胫骨骨折大鼠骨折愈合的影响.方法:120只SD大鼠随机分成3组:胫骨骨折组(F组,n=40),T10 脊髓完全性损伤合并胫骨骨折组(FS组,n=40),T10 脊髓完全性损伤合并胫骨骨折应用神经生长因子治疗组(FSN组,n=40).FS组及FSN组大鼠建立T10脊髓完全横断伴胫骨骨折模型,F组制备单纯胫骨骨折,伤后第4周对3组大鼠骨折断端行断层CT扫描,测量骨折断端最大横截面并计算骨痂灰度值;行生物力学三点弯曲试验;行骨组织形态计量学检查;骨密度测定;骨痂组织形态学观察;免疫组化法测定骨痂组织中骨钙素(OCN)的表达;观察3组大鼠骨痂中成骨细胞的超微结构变化;Western印迹检测Ⅰ、Ⅱ型胶原的表达.结果:F组骨痂断面最大直径明显小于FS组(P＜0.01)和FSN组(P＜0.05),FSN组小于FS组(P＜0.05);骨痂灰度值结果类似.三点折弯试验、骨计量学观察均发现F组各项生物力学参数优于其他2组(P＜0.05);FSN组优于FS组(P＜0.05).F组大鼠胫骨骨痂中OCN及Ⅰ型胶原蛋白表达明显高于其他2组(P＜0.05),FSN组高于FS 组(P＜0.05);而F组骨痂Ⅱ型胶原蛋白的表达明显低于FS组及FSN(P＜0.05),FSN组亦高于FS组(P＜0.05).形态学及超微结构观察见F组骨折愈合最佳,FSN组优于FS组.结论:失神经营养会影响骨折断端的骨化;神经生长因子对失神经状态下骨折断端的骨化有促进作用.     

神经生长因子促进大鼠胫骨骨折愈合的实验研究

目的:观察神经生长因子对大鼠胫骨骨折愈合以及胰岛素样生长因子表达的影响。方法:建立标准大鼠胫骨骨折模型,分别给予生理盐水和神经生长因子,术后1、2、4周分别进行X线骨痂评定、胫骨湿重称量以及半定量RT-PCR检测胰岛素样生长因子-1mRNA的表达。结果:术后4周时,神经生长因子组骨痂量和胫骨湿重明显多于对照组;术后1周、2周时神经生长因子组的胰岛素样生长因子-1mRNA的表达明显高于对照组。结论:应用神经生长因子可以促进大鼠胫骨各组愈合,同时促进骨折愈合过程中IGF-1表达。     

骨髓间充质干细胞对骨质疏松性骨折大鼠愈合的影响

目的探讨骨髓间充质干细胞(BMSCs)对骨质疏松性骨折大鼠愈合的影响。方法选取60只3月龄雌性SD大鼠,随机分为假手术组、模型组和MSCs组,每组20只,均采用切除卵巢去势法建立骨质疏松模型组,模型组和MSCs组大鼠全部制作胫骨骨折模型。分别于术后第1、14和21天给予MSCs组大鼠MSCs移植,假手术组和模型组大鼠给予同等体积的生理盐水注射。于细胞移植后2、4和6周,取骨痂,采用PCR法测定骨形成保护蛋白-2(BMP-2)、血管内皮生长因子(VEGF)、骨保护素(OPG)和核因子κB受体激活物配体(RANKL)的mRNA表达水平;评估移植8周后骨折愈合情况、骨折侧胫骨骨密度及生物力学参数。结果与假手术组比较,模型组大鼠BMP-2、VEGF、OPG和RANKL的mRNA表达水平显著下降,OPG/RANKL比值降低(P0.05);与模型组比较,MSCs组大鼠BMP-2、VEGF、OPG和RANKL的mRNA表达水平显著升高,OPG/RANKL比值增高(P0.05)。与模型组比较,MSCs组大鼠骨折愈合明显增加(P0.05),骨密度显著提高(P0.05),生物力学参数(最大载荷、弹性载荷、刚度和最大挠度)显著改善(P0.05)。结论间充质干细胞能够增加骨质疏松性骨折大鼠骨密度,促进骨组织再生,加快骨折愈伤组织的生长,对大鼠骨折的愈合具有促进作用,为临床治疗骨质疏松性骨折提供了指导作用。     

雷尼酸锶对骨质疏松股骨折愈合影响的实验研究

目的探讨雷尼酸锶对骨质疏松股骨折愈合的影响。方法选用成年雌性SD大鼠40只,采用双侧卵巢切除和骨折手术方法建立骨质疏松股骨折(OPF)模型。将大鼠分为对照组和实验组,实验组给予雷尼酸锶(Strontium ranelate,SR),对照组给予同体积生理盐水。X线观察骨折断端愈合情况,检测二组大鼠股骨骨密度(BMD),HE染色观察骨痂组织,免疫组织化学方法检测骨形态发生蛋白-2(BMP-2),抗酒石酸酸性磷酸酶染色检测破骨细胞数。结果①OPF术后6周实验组骨折断端骨痂较大,有连续骨痂通过骨折断端;对照组骨折断端骨痂较小;②OPF术后2周,实验组BMP-2表达阳性细胞数高于对照组(P<0.05)。术后6周,实验组较对照组骨折愈合明显,BMP-2表达高于对照组(P<0.05);③OPF术后6周时,实验组骨小梁面积比和骨小梁平均宽度高于对照组,骨小梁平均间隔小于对照组(P<0.05)。④实验组破骨细胞数量减少(P<0.05)。⑤OPF术后2、6周,实验组BMD高于对照组(P<0.05)。结论研究结果提示雷尼酸锶可促进骨质疏松股骨折愈合,增加骨密度,抑制破骨细胞。     

降钙素基因相关肽对大鼠胫骨骨折早期愈合的影响

目的 建立大鼠单纯胫骨骨折模型,分别给予大鼠腹腔注射不同浓度的外源性降钙素基因相关肽(calcitonin gene-related peptide,CGRP)及其特异性受体拮抗剂,评价并探讨CGRP对骨折早期愈合的影响及其对骨形成蛋白-2(bone morphogenetic protein-2,BMP-2)的表达.方法 制作单纯胫骨骨折模型,将SD大鼠应用随机数字表法随机分为3组,每组10只,分别给予腹腔注射生理盐水(对照组)、CGRP及CGRP受体拮抗剂.术后1、2周和4周时取材,进行骨折部骨痂测量,HE染色观察骨折局部形态变化;利用免疫组织化学检测骨组织中BMP-2的表达情况.结果 大鼠腹腔内注射CGRP1周时,各组均可见到少许骨痂,组间未见明显差别;术后2周时,各组外观有差别,CGRP组大鼠胫骨骨折部呈膨大样,大鼠骨痂量多于盐水对照组和拮抗剂组;术后4周CGRP组骨折部膨大明显缩小,骨皮质连续性好,各组外形已接近正常胫骨形态及大小.免疫组织化学结果显示:CGRP组胫骨骨折大鼠在骨折断端附近BMP-2的表达信号强于对照组和拮抗剂组.用药1周时,BMP-2免疫活性细胞从对照组的(18.52±10.08)增加为(36.80±11.18),增加了近2.2倍,拮抗剂组则减少为(14.50±8.80);用药2周时,BMP-2免疫活性细胞从对照组的(20.26±6.78)增加为(52.18±10.42),拮抗剂组减少为(11.21±8.40),变化幅度最大;用药4周时,BMP-2免疫活性细胞的增加从对照组的(21.25±8.65)增加为(31.41±9.18),拮抗剂组则减少为(18.32±10.28).结论 CGRP可能具有促进大鼠胫骨骨折早期愈合的作用,CGRP可以诱导骨组织中BMP-2的表达,推测CGRP的成骨作用与BMP-2的表达具有相关性.     

三七总皂甙对实验性骨折愈合过程中低氧诱导因子-1αmRNA表达的影响

目的:探讨低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)在骨折愈合过程中的表达及三七总皂甙(total saponins of panax notoginseng,PNS)的干预作用。方法:在36只雄性SD大鼠左桡骨形成骨折模型,后随机分成对照组和PNS组,予以PNS100mg/kg/d腹腔内注射,对照组予以等量盐水。在骨折后7,14,21天X线观察骨折愈合及骨痂形成情况;HE染色了解骨痂形成情况;RT-PCR方法检测骨痂组织HIF-1αmRNA的表达。结果:X线显示PNS组骨痂形成明显加快,HE染色显示骨折断端成骨细胞数目及骨痂增多;对照组HIF-1αmRNA7天表达较低、14天达峰值、21天下降;PNS组7天、14天时点表达高于对照组(P0.05),21天低于对照组P0.05)。结论:PNS可上调骨折愈合过程中骨痂组织HIF-1α的表达,改善骨折部位的血供,促进骨折愈合。     

骨基质蛋白在小鼠胫骨牵引成骨过程中的表达

目的探讨小鼠胫骨牵引成骨动物模型,并检测骨基质蛋白在牵引成骨过程中的表达和作用.方法8周龄雄性CD-1小鼠36只,接受左胫骨中上段骨干横行截骨,安置特制延长外固定架,胫骨牵引过程包括5 d静止期,12 d牵引期和70 d固塑期,牵引速率为0.1 mm/Bid,共0.2 mm/d.术后于不同时间点分组采集左胫骨标本,分别作组织学检查和骨基质蛋白mRNA检测.结果组织学检查显示静止期其修复过程基本与骨折愈合相似.牵引期,被牵引骨痂显示三个典型的生物学功能区:纤维间区,初始骨基质前沿和微脊柱形成区.固塑期早期,牵引骨痂骨性愈合,第10周骨髓腔再通,新生骨再塑基本上完成.mRNA分析显示在牵引成骨早期,软骨内成骨及膜内成骨同时发生.牵引中期以后软骨内成骨逐渐减少,而转为以膜内成骨为主的成骨过程.结论通过对骨基质蛋白及相关蛋白的分析,结果表明牵引成骨与骨折愈合过程不同.早期软骨内成骨和膜内成骨同时存在,但在机械牵张力作用下转为以膜内成骨为主的成骨过程.该研究亦证实小鼠胫骨牵引成骨过程与人体及其他动物模型基本相同,可作为一种非常有意义的研究骨再生和修复的在体动物模型.     

基于VEGF/VEGFR-2信号通路研究淫羊藿苷促进大鼠胫骨干骨折愈合的作用机制

[目的]基于血管内皮生长因子/血管内皮生长因子受体-2(vascular endothelial growth factor/vascular endothelial growth factor receptor-2,VEGF/VEGFR-2)信号通路研究淫羊藿苷(icariin,ICA)对大鼠胫骨干骨折愈合的影响。[方法]将41只胫骨干骨折模型SD大鼠随机分为对照组,ICA低、高剂量组,每组各10只,另设阳性药物组,大鼠11只。ICA低、高剂量组以40、80mg/(kg·d)的ICA腹腔注射,阳性药物组以0.4mL复方骨肽注射液腹腔注射。对照组以等量0.9%氯化钠溶液腹腔注射,各组均为1次/d,连续21d。干预第7、14、21天进行微计算机断层扫描技术(micro-computed tomography,Mirco-CT)活体扫描,计算样本骨体积分数(bone volume/total volume,BV/TV),骨小梁数量(trabecular number,Tb.N)。脱颈处死大鼠,进行下肢生物力学测试,采用苏木精-伊红(hematoxylin-eosin,HE)染色观察骨痂组织血管新生情况,实时荧光定量聚合酶链式反应(quantitative Real-time polymerase chain reaction,QRT-PCR)检测骨痂组织VEGF、VEGFR-2 mRNA相对表达量,Western blot检测骨痂组织VEGF、VEGFR-2蛋白相对表达量。[结果]与对照组比较,ICA低、高剂量组、阳性药物组BV/TV更高,Tb.N更多,最大载荷更大,新生血管数量增多,面积也更大,VEGF、VEGFR-2 mRNA及蛋白相对表达量较高(P<0.05);与ICA低剂量组比较,ICA高剂量组、阳性药物组BV/TV更高,Tb.N更多,最大载荷更大,新生血管数量增多,面积也更大,VEGF、VEGFR-2 mRNA及蛋白相对表达量较高(P<0.05);与ICA高剂量组比较,阳性药物组BV/TV更高,Tb.N更多,最大载荷更大,新生血管数量增多,面积也更大,VEGF、VEGFR-2 m RNA及蛋白相对表达量较高(P<0.05)。[结论]ICA可加快胫骨干骨折大鼠骨量新生,促进血管形成,增强抵抗外力冲击的能力,且该效应呈剂量依赖性,推测可能与上调VEGF、VEGFR-2表达有关。     

大鼠海马HIF-1&agr;和EPO在衰老过程中的表达

目的:探讨大鼠海马组织中低氧诱导因子-1&agr;（HIF-1&agr;)和促红细胞生长素（EPO)在衰老过程中的表达规律及二者在神经系统衰老过程中的作用。方法:应用尼氏染色和免疫组织化学技术观察不同月龄组（3,18,24,30月)大鼠海马CA1区神经细胞尼氏体及HIF-1&agr;和EPO的表达情况。结果:随大鼠月龄的增大,海马CA1区神经细胞体积变大,排列稀疏,胞浆内尼氏体减少。随月龄增加海马CA1区HIF-1&agr;阳性细胞数增加,差异有统计学意义（P<0.05);大鼠海马CA1区EPO表达随月龄增加呈抛物线变化,由3月龄到18月龄,阳性细胞个数的增加有统计学意义（P<0.05),由18月龄到30月龄,阳性细胞个数减少有统计学意义（P<0.05)。结论: HIF-1&agr;与EPO在大鼠海马的表达在中年期之前,呈平行递增变化,在中年期之后,二者呈分离变化,提示HIF-1&agr;活性下降和蛋白质合成功能的减退是衰老过程中EPO表达减少的主要原因,通过加强内源性HIF-1&agr;的活性和补充外源性EPO有可能减缓神经系统的衰老。     

Effects of fatty acid regulation on visfatin gene expression in adipocytes

Background The levels of long-term elevated serum or intracellular free fatty acid （FFA） induce insulin resistance associated with central obesity. The insulin-mimetic protein visfatin is preferentially produced by visceral adipose tissues and has been implicated in obesity and insulin resistance. To identify that FFA is capable of inducing insulin resistance and to clarify the role of FFA on visfatin, we examined the effect of monounsaturated FFA oleate （C 18： 1） and saturated FFA palmitate （C 16：0） on glucose transport and visfatin gene expression in cultured 3T3-L1 adipocytes or preadipocytes. Methods FFA-free DMEM/F12, 0.125 mmol/L, 0.5 mmol/1 and 1.0 mmol/L oleate or palmitate was added to cultured 3T3-L1 adipocytes or preadipocytes and incubated overnight. Glucose transport was assessed as 3H- 2-deoxy-glucose uptake. Total RNA was extracted and subjected to RT-PCR for the measurement of visfatin mRNA levels. Statistical comparisons between control group and other groups were performed with the two-tailed paired t test, and one-way ANOVA was used to compare the mean values among the groups. Results Insulin increased specific membrane glucose transport in 3T3-L1 preadipocytes. Upregulation was evident from 15 minutes to 1 hour exposure to insulin. However, after 6-hour exposure to insulin, there was a downregulation in the response to insulin. Dose response studies demonstrated that 2-deoxy glucose transport was increased by 336% at 50 nmol/L insulin （P〈0.01）, and reached a maximal effect at 100 nmol/L insulin （P〈0.01）. Oleate and palmitate treatment did not influence basal glucose transport （without insulin stimulation）, whereas insulin-stimulated glucose transport was inhibited after overnight oleate and palmitate treatment in preadipocytes and adipocytes. In 3T3-L1 preadipocytes, insulin resistance could be achieved at 0.125 mmol/L oleate or palmitate （P〈0.05, respectively）, and the inhibition was dose dependent. In adipocytes, the inhibition was noted at 0.5 mmol/L oleate or 1.0 mmol/L palmitate. Visfatin mRNA expression increased during differentiation more than 1.5-fold. Bovine serum albumin （BSA） did not influence visfatin mRNA expression compared with the control group. Dose-response studies demonstrated that addition of 0.125 mmol/L oleate and palmitate to 3T3-L1 adipocytes decreased visfatin mRNA expression significantly （78%, 77%, respectively, relative to untreated control, P〈0.05）, and further to 65% （relative to untreated control, P〈0.05） and 55% （relative to untreated control, P〈0.01） at 1.0 mmol/L FFA. Furthermore, the suppression on preadipocytes was similar to that of adipocytes, which reached a maximal reduction of 44% （oleate, P〈0.05） and 47% （palmitate, P〈0.05） at 1.0 mmol/L FFA. Conclusions Oleic acid and palmitic acid may induce insulin resistance in 3T3-L1 adipocytes and preadipocytes. Downregulation of visfatin mRNA may contribute to impair insulin sensitivity caused by oleate and palmitate.     

Alleviation of hypoxic pulmonary vascular structural remodeling by L-arginine

目的探讨L-精氨酸(L-Arg)对低氧性肺血管结构重建的干预作用及其可能机制.方法将18只Wistar大鼠配伍后随机分为对照组、低氧组和低氧+L-Arg组(共6个配伍组).以右心导管法测定肺动脉压力,并对大鼠肺组织标本进行显微结构观测和超微结构观察,同时以分光光度法测定血浆一氧化氮(NO)含量,并对肺组织以内皮素-1(ET-1) cRNA探针进行原位杂交,研究肺动脉内皮细胞ET-1 mRNA的表达.结果低氧组大鼠肺动脉平均压明显高于对照组(20.33±2.18?mm?Hg vs 15.38±1.05?mm?Hg, P<0.05).低氧后大鼠肺血管显微及超微结构发生明显改变,肺血管结构重建形成.同时低氧组大鼠血浆NO间接含量明显低于对照组 (P<0.05 ).低氧后肺动脉内皮细胞ET-1 mRNA表达明显增强.然而,低氧+L-Arg组大鼠PAMP较低氧组明显降低(16.73±1.35?mm?Hg vs 20.33±2.18?mm?Hg, P<0.05).L-Arg缓解了低氧性肺血管结构重建的形成.同时低氧+L-Arg组大鼠血浆NO间接含量明显高于低氧组(P<0.05).L-Arg使低氧大鼠肺动脉内皮细胞ET-1 mRNA表达明显受抑制.结论 L-Arg对低氧性肺血管结构重建以及低氧性肺动脉高压的形成有重要的调节作用,其机制可能与通过促进低氧大鼠体内NO生成,从而抑制肺动脉内皮细胞ET-1 mRNA表达有一定的关系.     

PPARγ1基因转染对心肌缺血再灌注后的影响和意义

目的  探讨心肌细胞过表达过氧化物酶体增殖物激活受体γ1（PPARγ1）基因后对缺血再灌注损伤导致的血流动力学、心肌梗死（心梗）面积、血清肌钙蛋白I（cTnI）及心肌基质金属蛋白酶-9（MMP-9）浓度的变化及意义。方法  30只SD大鼠随机分成3组（n =10）：SHAM组、MIRI组及PPARγ1组。SHAM组和MIRI组开胸经冠状动脉（冠脉）转染携带绿色荧光蛋白的腺病毒载体（Ad-EGFP）,PPARγ1组转染携带PPARγ1基因的腺病毒载体（Ad-PPARγ1）至心肌组织。稳定3 d后重新开胸,SHAM组只过线,不接扎;MIRI组及PPARγ1组结扎冠脉左前降支30 min,再灌注120 min。观察缺血前（T0）、缺血30 min（T1）、再灌注30 min（T2）、再灌注120 min（T3）时的心率（HR）、平均动脉压（MAP）、左室收缩压（LVSP）、左室舒张末压（LVEDP）,左室压最大上升和下降速率（±dp/dt max）;再灌注120 min时检测心梗面积、血清心肌肌钙蛋白I（cTnI）和组织MMP-9浓度的变化。结果  与T0时比较,T1～T3时MIRI组、PPARγ1组LVEDP升高,HR减慢,LVSP和（±dp/dt max）降低,MAP除PPARγ1组在T3时差异无统计学意义,也明显降低（P <0.05）;与SHAM组比较,T1～T3时MIRI组、PPARγ1组LVEDP升高,HR减慢,LVSP和（±dp/dt max）降低,MAP除PPARγ1组在T3时差异无统计学意义,也明显降低（P <0.05）;与MIRI组比较,PPARγ1组±dp/dt max升高,LVSP在T2、T3时升高,MAP在T3时升高（P <0.05）;SHAM组无心肌梗死,MIRI组和PPARγ1组的缺血面积差异无统计学意义（P >0.05）,而MIRI组和PPARγ1组心肌梗死面积、cTnI和MMP-9均高于SHAM组,PPARγ1组低于MIRI组（P < 0.05）。结论  过表达PPARγ1基因能通过减轻缺血再灌注过程中血流动力学紊乱、减少心梗面积、降低血清cTnI及组织MMP-9的浓度,从而起到保护心肌的作用。

     

不同血液透析对患者矿物质骨异常影响分析

目的：探讨分析不同血液透析对患者矿物质骨异常的影响。方法：选择我院收治的慢性肾功能衰竭且维持性血液透析12个月以上患者106例,根据患者透析方式将其分为A组43例、B组3例、C组33例,A组给予低通量透析、B组给予高通量透析、C组给予高通量透析滤过,观察时间6个月,分析三组患者血磷、血钙、全段甲状旁腺素（iPTH）、I型前胶原氨基末端肽（PINP）、成纤维生长因子23（FGF-23）、骨质疏松患病率等指标变化情况。结果：A组患者治疗6个月Ca、P、iPTH、PINP、FGF-23水平均无明显变化（P&amp;gt;0.05）;B组患者治疗6个月iPTH水平显著降低（P&amp;lt;0.05）,Ca、P、PINP、FGF-23水平无明显变化（P&amp;gt;0.05）;C组患者治疗6个月iPTH、PINP、FGF-23水平显著降低（P&amp;lt;0.05）,Ca、P水平无明显变化（P&amp;gt;0.05）。组间比较,B组、C组治疗6个月iPTH水平明显低于A组（P&amp;lt;0.05）,治疗6个月C组PINP、FGF-23水平明显低于A组、B组（P&amp;lt;0.05）。治疗6个月三组患者骨质疏松患病率与入组时比较均无明显变化（P&amp;gt;0.05）,且三组骨质疏松患病率比较无明显差异（P&amp;gt;0.05）。透析6个月后,三组血清钙、磷以及钙磷乘积达标率均无显著差异（P&amp;gt;0.05）,C组iPTH达标率显著高于A组（P&amp;lt;0.05）。结论：不同血液透析方式对维持性血液透析患者矿物质骨代谢影响有所不同,高通量血液透析与低通量透析相比较,更有利于患者矿物质骨代谢指标的改善,且高通量血液透析滤过在骨代谢指标改善方面更具优势。     

Inhibitory effect of tea polyphenols on renal cell apoptosis in rat test subjects suffering from cyclosporine-induced chronic nephrotoxicity

Objective To investigate the inhibitory effect of tea polyphenols on renal cell apoptosis in rat test subjects suffering from cyclosporine A (CsA)-induced chronic nephrotoxicity.Methods Four groups of rats with CsA-induced chronic nephrotoxicity were respectively treated with vehicle olive oil, tea polyphenols, CsA and tea polyphenols plus CsA. At the end of the 28th day of treatment, 24 hours urine and blood samples were obtained, and the animals were then sacrificed. The serum and urine samples were analysed for creatinine clearance, and kidney tissue was used for pathologic analysis of renal tubular injury and interstitial fibrosis. The TUNEL assay, apoptosis-related enzyme caspase-3 mRNA detected by RT-PCR, and its enzymatic activity were analysed for the possible detections of cell apoptosis.Results CsA-treated rats displayed increased apoptosis of the tubular and interstitial cells, in comparison with vehicle-treated controls (18. 3±4. 6 vs 4. 8±1.3 cells/mm2, P < 0. 05 ) . In comparision with a     

Effects of bone marrow-derived endothelial progenitor cell transplantation on vein microenvironment in a rat model of chronic thrombosis

Background  Endothelial progenitor cells (EPCs) have been used in both experimental studies and clinical treatments of limb ischemia, as well as in the construction of engineered vascular tissue. The objective of this study was to investigate the effects of transplanted bone marrow-derived EPCs on the vein microenvironment in a rat model of chronic vein thrombosis.
Methods  Mononuclear cells were isolated from the bone marrow of immature rats by density gradient centrifugation, cultured, and then transplanted into experimentally induced thrombi into inferior vena cava through the femoral vein. Vascular endothelial growth factor (VEGF), angiopoietin-1 (ANG-1)  and monocyte chemotactic protein-1 (MCP-1) mRNA and protein expression levels were measured by real-time quantitative polymerase chain reaction and Western blotting of thrombi and adjacent caval walls 28 days post-transplantation.
Results  Levels of VEGF, ANG-1, and MCP-1 mRNA in EPC-transplanted thrombi were 100%, 230.7%, and 212.5% of levels detected in the sham-operated group (P<0.01), and 99.9%, 215.4%, and 177.8% of levels detected in the experimental control group (P< 0.01). VEGF, ANG-1 and MCP-1 protein levels exhibited a similar trend.
Conclusions  Transplanted bone marrow-derived EPCs appear to alter the vein microenvironment in experimentally induced chronic vein thrombosis by upregulating cytokines associated with thrombic organization and recanalization.

     

Expression and function of c-kit receptor in bone marrow mononuclear cells of patients with myelodysplastic syndromes

目的　了解骨髓增生异常综合征 (MDS)骨髓单个核细胞c kit受体的表达与功能。方法　采用免疫荧光方法测定c kit受体蛋白 (CD117)的表达 ;逆转录 聚合酶链反应方法测定c kitmRNA的表达 ;细胞培养检测c kit受体的功能。结果　CD117表达率正常人为 3 0 4 %± 1 4 9% ,MDS患者为 8 58%± 5 2 8% ,两者差异有显著性 (P <0 0 5) ;RA患者为 5 12 %± 2 13% ) ,RAEB RAEB t患者为 10 0 1%± 5 0 7% ,两者差异有显著性 (P <0 0 5) ;MDS继发白血病患者为 32 4 3%± 18 16 %。MDS患者c kit基因mRNA表达与CD117表达相一致。正常骨髓单个核细胞体外半固体培养 ,在加入造血干细胞因子、白细胞介素 3、红细胞生成素 (SCF IL 3 Epo)后形成的粒 巨噬细胞集落 (CFU GM)较仅加入IL 3 Epo显著增多 (P <0 0 5) ,红系爆式集落 (BFU E)数量极显著增多 ,且BFU E体积显著增大(P <0 0 1)。MDS患者在上述相同条件下CFU GM数量无明显变化 (P >0 0 5) ,与正常对照比较 ,CFU GM和BFU E数量均显著减少 (P <0 0 5)。结论　MDS患者骨髓单个核细胞CD117表达明显高于正常对照 ,且与病情进展相关 ;c kitmRNA表达与CD117表达相一致 ;MDS患者骨髓单个核细胞体外培养时 ,SCF协同IL 3、Epo促进造血干 祖细胞增殖分化能力较正常对照明     

Effects of chronic multiple stress on learning and memory and the expression of Fyn, BDNF, TrkB in the hippocampus of rats

Background The effect of chronic stress on cognitive functions has been one of the hot topics in neuroscience. But there has been much controversy over its mechanism. The aim of this study was to investigate the effects of chronic multiple stress on spatial learning and memory as well as the expression of Fyn, BDNF and TrkB in the hippocampus of rats. Methods Adult rats were randomly divided into control and chronic multiple stressed groups. Rats in the multiple stressed group were irregularly and alternatively exposed to situations of vertical revolution, sleep expropriation and restraint lasting for 6 weeks, 6 hours per day with night illumination for 6 weeks. Before and after the period of chronic multiple stresses, the performance of spatial learning and memory of all rats was measured using the Morris Water Maze （MWM）. The expression of Fyn, BDNF and TrkB proteins in the hippocampus was assayed by Western blotting and immunohistochemical methods. The levels of Fyn and TrkB mRNAs in the hippocampus of rats were detected by RT-PCR technique. Results The escape latency in the control group and the stressed group were 15.63 and 8.27 seconds respectively. The performance of spatial learning and memory of rats was increased in chronic multiple stressed group （P〈0.05）. The levels of Fyn, BDNF and TrkB proteins in the stressed group were higher than those of the control group （P〈0.05）. The results of immunoreactivity showed that Fyn was present in the CA3 region of the hippocampus and BDNF positive particles were distributed in the nuclei of CA1 and CA3 pyramidal cells as well as DG granular cells. Quantitative analysis indicated that level of Fyn mRNA was also upregulated in the hippocampus of the stressed group （P〈0.05）. Conclusions Chronic multiple stress can enhance spatial learning and memory function of rats. The expression of Fyn, BDNF and TrkB proteins and the level of Fyn mRNA are increased in the stessed rat hippocampus. These suggest that Fyn and BDNF/TrkB signal transduction pathways may participate in the process of the enhanced learning and memory durina chronic multiple stress.