氟对大鼠腰椎骨组织形态计量学的影响

Objective To study the effect of fluorine on the bone histomorphometry of humbar in rats.Methods Ninety 2-month-old SPF Sparague-Dawley rats,half male and female,were randomly divided into 9 groups:control[(childhood(CS),adult(AS),long-time(NS)]group and drug group[childhood high-fluoride and low-fluoride group(CHS,CLS),adult high-fluoride and low-fluoride(AHS,ALS),long-term high-fluoride and low-fluoride(CLHS,CLLS)].The control group was administered orally with solution of 0.9%NaCl,while the drug group was given orally with different dose of NaF at the same time. Sections of the fifth lumbar were made which was undecalicified for bone histomorphometric analysis, including the percentage of trabecular bone area (% Tb.Ar),trabecular thickness(Tb.Th), trabecular number(Tb.N), trabecular separation(Th.Sp) ; broken trabecular bone area cells (Oc.N), osteoclast perimeter percentage (% Oc.Pm), the percentage of labeled perimeter (% L.Pm), bone mineral apposition rate(MAR), osteoblast perimeter(Ob.PM), trabecular bone perimeter formation rate (BFR/BS),trabecular bone area formation rate (BFR/BV), the total area of bone formation rate (BFR/TV). Results [1]The percentage of Tb.Ar, Tb.Th, Tb.N,%L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of CHS group [(50.63 ±7.44)%, (150.26 ± 27.51 )μm, (3.44 ± 0.47)N/mm, (50.63 ± 7.44)%, (0.85 ± 0.03)μm/d, (8.45 ± 2.36)μm/d ×100, (381.16 ± 41.62)%/year, (75.07 ± 4.81)%/year] was higher than that of CS group [(29.71 + 9.32)%,(110.93 ± 28.19)μm, (2.68 ± 0.34)N/mm, (24.00 ± 1.22)%, (0.65 ± 0.03)μm/d, (5.43 ± 0.18)μm/d × 100,(141.32 ± 9.29)%/year, (58.14 ± 2.3)%/year, all P < 0.05)]. The %Tb.Ar, Tb.Th, %L.Pm, MAR, BFR/BS,BFR/BV, BFR/TV and Ob.PM of CLS group [(40.76 ± 6.43)%, (164.25 ± 45.65)μm, (42.02 ± 6.12)%, (0.85 ±0.04)μm/d, (8.95 ± 3.73)μm/d × 100, (378.73 ± 35.39)%/year, (73.52 ± 8.71)%/year, (1.41 ± 0.05)μm] were increased (all P < 0.05). [2]Compared with AS group, the %Tb.Ar,Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS,BFR/BV and BFR/TV of AHS group[ (50.62 ± 5.76)%, (0.51 ± 0.05)N/mm, (1.13 ± 0.05)%, (42.3 ± 7.02)%,(1.28 ± 0.09)μm/d, (12.91 ± 1.52)μm/d × 100, (390.12 ± 43.56)%/year, (65.21 ± 22.13)%/year] was higher than that of AS group[ (42.73 ± 5.22)%, (0.41 ± 0.17)N/ram, (0.77 ± 0.52)%, (28.43 ± 6.93)%, (0.80 ± 0.03)μm/d, (9.83 ± 1.44)μm/d × 100, (324.43±53.44)%/year and(48.35 ± 9.36)%/year, all P < 0.05)] . The %Tb.At, Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of ALS group [(51.14 ± 6.22)%, (0.49 ±0.61)N/mm, (1.17 ± 0.11)%, (45.06 ± 6.92)%, (1.39 ± 0.08)μm/d, (12.87 ± 1.35)μm/d × 100, (394.6 ±50.23)%/year and(66.31 ± 18.93)%/year] were higher than that of AS group(P < 0.05) .[3] The Ob.PM ,Oc.N and %Oc.Pm of CLHS group[ (1.47 ± 0.27)μm, (0.58 ± 0.13)N/mm, (1.14 ± 0.07)%] were obviously increased(P <0.05), as compared with NS group [ (0.82 ± 1.20)μm, (0.42 ± 0.25)N/mm and (0.75 ± 0.64)%, all P < 0.05].Conclusions The short-term administration of NaF on rats in the growing period increases the bone formation and osteoblast activities of young rats and adult rats. The long-term administration of NaF on rats does not increase the bone formation of rats in growth period. The osteoblast activities as well as the bone absorption of lumbar vertebra were strengthened. The likelihood of bone fracture became larger. The negative effects on bone metabolism and bone quality of rats were gradually displayed along with the prolongation of sodium fluoride usage.     

氟对大鼠腰椎骨组织形态计量学的影响

Objective To study the effect of fluorine on the bone histomorphometry of humbar in rats.Methods Ninety 2-month-old SPF Sparague-Dawley rats,half male and female,were randomly divided into 9 groups:control[(childhood(CS),adult(AS),long-time(NS)]group and drug group[childhood high-fluoride and low-fluoride group(CHS,CLS),adult high-fluoride and low-fluoride(AHS,ALS),long-term high-fluoride and low-fluoride(CLHS,CLLS)].The control group was administered orally with solution of 0.9%NaCl,while the drug group was given orally with different dose of NaF at the same time. Sections of the fifth lumbar were made which was undecalicified for bone histomorphometric analysis, including the percentage of trabecular bone area (% Tb.Ar),trabecular thickness(Tb.Th), trabecular number(Tb.N), trabecular separation(Th.Sp) ; broken trabecular bone area cells (Oc.N), osteoclast perimeter percentage (% Oc.Pm), the percentage of labeled perimeter (% L.Pm), bone mineral apposition rate(MAR), osteoblast perimeter(Ob.PM), trabecular bone perimeter formation rate (BFR/BS),trabecular bone area formation rate (BFR/BV), the total area of bone formation rate (BFR/TV). Results [1]The percentage of Tb.Ar, Tb.Th, Tb.N,%L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of CHS group [(50.63 ±7.44)%, (150.26 ± 27.51 )μm, (3.44 ± 0.47)N/mm, (50.63 ± 7.44)%, (0.85 ± 0.03)μm/d, (8.45 ± 2.36)μm/d ×100, (381.16 ± 41.62)%/year, (75.07 ± 4.81)%/year] was higher than that of CS group [(29.71 + 9.32)%,(110.93 ± 28.19)μm, (2.68 ± 0.34)N/mm, (24.00 ± 1.22)%, (0.65 ± 0.03)μm/d, (5.43 ± 0.18)μm/d × 100,(141.32 ± 9.29)%/year, (58.14 ± 2.3)%/year, all P < 0.05)]. The %Tb.Ar, Tb.Th, %L.Pm, MAR, BFR/BS,BFR/BV, BFR/TV and Ob.PM of CLS group [(40.76 ± 6.43)%, (164.25 ± 45.65)μm, (42.02 ± 6.12)%, (0.85 ±0.04)μm/d, (8.95 ± 3.73)μm/d × 100, (378.73 ± 35.39)%/year, (73.52 ± 8.71)%/year, (1.41 ± 0.05)μm] were increased (all P < 0.05). [2]Compared with AS group, the %Tb.Ar,Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS,BFR/BV and BFR/TV of AHS group[ (50.62 ± 5.76)%, (0.51 ± 0.05)N/mm, (1.13 ± 0.05)%, (42.3 ± 7.02)%,(1.28 ± 0.09)μm/d, (12.91 ± 1.52)μm/d × 100, (390.12 ± 43.56)%/year, (65.21 ± 22.13)%/year] was higher than that of AS group[ (42.73 ± 5.22)%, (0.41 ± 0.17)N/ram, (0.77 ± 0.52)%, (28.43 ± 6.93)%, (0.80 ± 0.03)μm/d, (9.83 ± 1.44)μm/d × 100, (324.43±53.44)%/year and(48.35 ± 9.36)%/year, all P < 0.05)] . The %Tb.At, Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of ALS group [(51.14 ± 6.22)%, (0.49 ±0.61)N/mm, (1.17 ± 0.11)%, (45.06 ± 6.92)%, (1.39 ± 0.08)μm/d, (12.87 ± 1.35)μm/d × 100, (394.6 ±50.23)%/year and(66.31 ± 18.93)%/year] were higher than that of AS group(P < 0.05) .[3] The Ob.PM ,Oc.N and %Oc.Pm of CLHS group[ (1.47 ± 0.27)μm, (0.58 ± 0.13)N/mm, (1.14 ± 0.07)%] were obviously increased(P <0.05), as compared with NS group [ (0.82 ± 1.20)μm, (0.42 ± 0.25)N/mm and (0.75 ± 0.64)%, all P < 0.05].Conclusions The short-term administration of NaF on rats in the growing period increases the bone formation and osteoblast activities of young rats and adult rats. The long-term administration of NaF on rats does not increase the bone formation of rats in growth period. The osteoblast activities as well as the bone absorption of lumbar vertebra were strengthened. The likelihood of bone fracture became larger. The negative effects on bone metabolism and bone quality of rats were gradually displayed along with the prolongation of sodium fluoride usage.     

盐酸吡格列酮对SHR心脏成纤维细胞增殖的影响及其与一氧化氮的关系

目的　研究胰岛素增敏剂盐酸吡格列酮 (PIO)对培养的自发性高血压大鼠 (SHR)和试验大鼠 (Wistar)心脏成纤维细胞 (CFs)增殖的影响及其与一氧化氮 (NO)的关系。方法　采用胶原酶消化法培养SHR和Wistar的CFs ,四氮唑蓝比色法 (MTT)测定CFs的增殖状况 ,硝酸还原酶法测定CFs培养上清NO浓度。结果　 (1) 1× 10 - 7、1× 10 - 6 、5× 10 - 6 、1× 10 - 5 mol/L的PIO作用 2 4h ,SHR和Wistar的吸收值 (A4 90 值 )分别为SHR 0 .19± 0 .0 1、0 .18± 0 .0 1、0 .16± 0 .0 1、0 .16± 0 .0 2 ;Wistar 0 .2 1± 0 .0 1、0 .19± 0 .0 1、0 .18± 0 .0 1、0 .17± 0 .0 1,与各自对照组 (SHR 0 .2 2± 0 .0 1,Wistar 0 .2 1± 0 .0 2 )相比 ,差异非常显著 (P <0 .0 1)。 (2 ) 5× 10 - 6 mol/L的PIO作用 12h、2 4h、3 6h、48h、60h、72h后 ,SHR的A490 值与同一时间点相比 ,差异非常显著 (P <0 .0 0 1) ;Wistar的A4 90 值与同一时间点的对照组相比 ,差异非常显著 (P <0 .0 0 1)。 (3 ) 5×10 - 6 mol/L的PIO干预 48小时后 ,SHR和Wistar的CFs培养上清NO浓度分别为 111.2± 12 .4μmol/L、2 2 1.7± 3 5.3μmol/L ,与各自对照组 (76.8± 2 .4μmol/L、112 .1± 8.9μmol/L)相比 ,差异非常显著 (P <0 .0 0 1)。 结论     

电应激对十二指肠组织形态及氧化反应体系的影响

目的 观察电应激对十二指肠组织形态及功能代谢的影响，旨在探讨多种疾病过程中肠病理学及病理生理学改变的一般机制。方法 用电刺激复制应激模型，图象计量分析的方法进行组织学观察，化学方法检测组织内SOD，MDA和NO含量。结果 应激组平滑肌及肠腺细胞核的面积、周长、最大直径、最小直径分别是(56．1±98．4)μm2，(115，4±69.6)μm2；(25．1±50.4)μm2，(47.1±60．8)μm2；(6．0±7．50)μm2，(5．5±7．0)μm2；(1．3±2．5)μm2，(1.0±1．8)μm2，与对照组相比均有显著性改变；组织内SOD，MDA和NO含量分别是(113．5±3．6)NU/mg蛋白，(1．93±O．15)mol/g蛋白和(0，313±0．086)μmol/g蛋白，较对照组明显增高(P     

乳球菌分泌的白介素-12对哮喘小鼠的疗效观察

目的将小鼠白介素-12基因(mIL-12)通过质粒载体pSEC导入乳酸乳球菌(NZ9000)中,观察mIL-12在细菌中的表达;通过鼻腔免疫来观察mIL-12对小鼠哮喘模型气道炎症及辅助T细胞亚群的影响。方法将mIL-12连接到pSEC载体上并将其转化到NZ9000中,用乳链菌肽(Nisin)诱导其表达IL-12蛋白质并做鉴定,将菌液离心浓缩后给小鼠滴鼻。BALB/c小鼠50只,随机分为5组,即对照组(A组);PBS组(B组);野生菌组(C组);rIL-12组(D组);工程菌组(E组)。除A组外,余各组每鼠在1、7、14 d腹腔注射卵白蛋白OVA(100μg/0.5ml),在20~26 d雾化吸入2%OVA,每天1次。B、C、E组在1、2、3、14、19、20、21 d分别给予10μl的PBS,不含质粒载体的野生乳酸菌(约5×108CFU),含IL-12的质粒载体的工程菌(约5×108CFU)。D组20、21、22天给3 ng/只重组白介素-12,第28天杀小鼠测支气管肺泡灌洗液(BALF)中的嗜酸粒细胞(EOS)个数及白介素-4(IL-4)、干扰素(IFN-γ)水平,肺组织HE染色切片观察病变。结果mIL-12能够在NZ9000中高效表达并释放到细胞外。A-E组BALF中EOS的数量分别为(0.51±0.23)×104cell/ml(、21.32±5.71)×104cell/ml(、22.84±6.16)×104cell/ml(、2.15±0.63)×104cell/ml、(2.62±0.73)×104cell/ml;IL-4的含量分别为68.43±9.08 pg/ml、332.13±61.36 pg/ml、324.65±57.27 pg/ml、121.25±35.55 pg/ml、114.22±15.70 pg/ml;IFN-γ的含量分别为110.28±12.19pg/ml、56.25±12.08 pg/ml、70.36±8.27 pg/ml、166.11±57.12 pg/ml、171.75±60.90 pg/ml。B和C组的EOS数I、L-4与IFN-γ含量与A组相比有统计学差异(P<0.01);D、E组与B、C组比较有统计学意义(P<0.01)。工程菌组小鼠肺组织炎症细胞浸润明显减少,气道上皮损害减轻。结论mIL-12在乳酸乳球菌中能高效表达;鼻腔应用mIL-12能减少全身应用的副作用,且经济实用并较好地调节Thl/Th2平衡,对小鼠哮喘气道炎症的保护较为理想。     

支气管哮喘大鼠气道重塑中肺泡巨噬细胞的作用

目的研究支气管哮喘(简称哮喘)大鼠气道重塑过程中肺泡巨噬细胞(AM)的变化及其作用。方法48只清洁级雄性幼年SD大鼠按随机数字表法分为正常对照组(A组)、哮喘3d组(B组)、哮喘14d组(C组)和哮喘30d组(D组),每组12只。应用鸡卵白蛋白(OVA)建立哮喘大鼠模型,纯化支气管肺泡灌洗液(BALF)中的AM,测定AM中肿瘤坏死因子α(TNF-α)、前列腺素E2(PGE2)的含量和基质金属蛋白酶9基因(MMP-9mRNA)及其组织抑制物1基因(TIMP-1mRNA)的表达,并测量大鼠支气管壁总面积和平滑肌面积,计算单位基底膜周径(Pbm)的支气管壁厚度(WAt)和平滑肌厚度(WAm)。结果D组WAt和WAm[(85±9)μm2/μm、(28.6±4.9)μm2/μm]与A组[(67±10)μm2/μm、(16.8±2.4)μm2/μm]比较差异有统计学意义(t值分别为2.938、3.227,P均<0.01);D组AM中TNF-α和PGE2含量[(0.68±0.25)μg/L、(0.122±0.030)μg/L]与A组[(0.37±0.09)μg/L、(0.079±0.018)μg/L]比较差异有统计学意义(t值分别为2.683、3.016,P均<0.01),与B组[(0.74±0.29)μg/L、(0.120±0.028)μg/L]、C组[(0.71±0.23)μg/L、(0.117±0.028)μg/L]比较差异无统计学意义(t值分别为1.624、0.472、0.935、0.533,P均>0.05);D组AM中MMP-9mRNA及TIMP-1mRNA含量吸光度(A)值分别为0.346±0.033、0.361±0.040,与C组(0.279±0.015、0.259±0.015)比较差异有统计学意义(t值分别为2.574、2.716,P均<0.01),D组(0.183±0.025)与B组(0.136±0.014)比较差异有统计学意义(t值分别为2.913、3.017,P均<0.01),D组(0.104±0.007)与A组(0.109±0.008)比较差异有统计学意义(t值分别为3.632、3.487,P均<0.01);各组AM中MMP-9mRNA含量与WAt、WAm呈正相关(r值分别为0.693、0.738,P均<0.01),AM中TIMP-1mRNA含量与WAt、WAm呈正相关(r值分别为0.823、0.876,P均<0.01)。结论哮喘大鼠AM及其分泌的一些细胞因子与气道重塑关系密切。     

自身免疫病患者外周血中B细胞激活因子基因表达水平的测定

目的　建立测定B细胞激活因子 (B lymphocytestimulator ,BlyS)mRNA含量的荧光定量反转录 聚合酶链反应 (RT PCR)法 ,并用来检测自身免疫病 [系统性红斑狼疮 (SLE)、类风湿关节炎 (RA) ]患者外周血单个核细胞 (PBMCs)中BlyS的基因表达水平 ,探讨BlyS基因表达水平与自身免疫性疾病发病机制的关系。方法　构建克隆载体 pMD18 T BlyS作为定量模板 ,基于TaqMan荧光探针技术 ,建立实时荧光RT PCR方法在GeneAmp 5 70 0型检测仪上定量检测 19例自身免疫病(SLE、RA)确诊病人、2 0例亚临床病人 (主要是抗核抗体阳性 )、8例其他对照性疾病患者 (自身抗体阴性 ,免疫球蛋白升高 )、2 0名正常健康献血者的外周血BlySmRNA表达含量。结果　 19例自身免疫病确诊病人的PBMCs中均有BlySmRNA的表达 ,范围从 9 7× 10 5～ 3 2× 10 8拷贝 /μgRNA ,均值为 (8 4± 7 9)× 10 7拷贝 /μgRNA ;2 0例亚临床病人的强度为 8 6× 10 4～ 3 8× 10 6拷贝 /μgRNA ,均值为 (1 3± 1 2 )× 10 6拷贝 /μgRNA ;2 0名正常健康人的强度为 5 5× 10 4～ 4 9× 10 5拷贝 /μgRNA ,均值为 (1 7± 1 4 )× 10 5拷贝 /μgRNA ;8例自身抗体阴性而免疫球蛋白升高的其他疾病患者的强度为 5 8× 10 5～ 3 5× 10 7拷贝 /μgRNA ,均值为 (1 2±     

维拉帕米对实验兔血管成形术后内膜增殖和血管重塑的影响

目的 :探讨维拉帕米对血管成形术后新生内膜增殖和血管重塑的影响及其机制。方法 :家兔 2 4只 ,随机分为假手术组、对照组和干预组。以球囊导管损伤家兔腹主动脉内膜 ,喂养 4周后 ,对后两组家兔腹主动脉狭窄部位行球囊成形术。其中对照组仅行血管成形术 ,干预组于术前 30min及术后每天给予维拉帕米肌注 [剂量 0 .5mg/ (kg·d) ]。分别于术前 ,术后 4 8h、1周及 4周采静脉血测定 3组家兔血清一氧化氮 (NO)含量。术后 4周处死动物 ,对成形术部位腹主动脉标本作病理形态学检查和免疫组化检查 ,并利用计算机图像分析系统分别测量血管管腔面积、新生内膜面积、内弹力板围绕面积、内膜 /中膜面积 ,计算增殖细胞核抗原 (PCNA)增殖指数(PI)。结果 :血管成形术后血清NO浓度降低 ,但术后 1周和 4周维拉帕米干预组血清NO浓度高于对照组同期水平 ,差异有非常显著性意义 (均 P <0 .0 1)。病理形态学检查示对照组和干预组管腔面积分别为 (6 .4 18±0 .6 5 8)和 (13.4 82± 1.2 0 5 )× 10 4μm2 ;内弹力板围绕面积分别为 (19.719± 1.0 87)和 (2 4 .32 6± 1.4 2 7)× 10 4μm2 。上述指标两组之间比较差异均有非常显著性意义 (均P <0 .0 1)。对照组和干预组新生内膜面积分别为 (13.176± 0 .6 34)和 (10 .195± 0 .5     

"血宝"对化学药物性骨髓损伤保护作用的体内实验研究

目的 :探讨“血宝”对化学药物性骨髓损伤的保护作用。方法 :昆明小鼠经大剂量环磷酰胺腹腔注射 ,约 5d后出现显著的骨髓损伤 ,外周血三系减少 ,骨髓造血功能低下。对骨髓损伤小鼠胃饲“血宝”10d后观察外周血三系细胞计数及骨髓有核细胞总数 ,并体外培养小鼠骨髓细胞 ,计数BFU E和CFU GM的集落产率。结果 :实验组外周血三系细胞数最低为WBC (3.5± 0 .9)× 10 9/L、RBC (4.7± 0 .4 )× 10 12 /L、PLT (46 7.9±4 3.3)× 10 9/L、HGB 92 .8± 4 .4 g/L ,骨髓有核细胞总数最低为 (5 .5± 0 .2 )× 10 6/股骨 ,其中RBC、PLT、HGB、骨髓有核细胞总数显著高于阴性对照组 [RBC (3.3± 0 .8)× 10 12 /L、PLT (2 39.8± 79.0 )× 10 9/L、HGB (49.2±10 .6 ) g/L、骨髓有核细胞总数 (1.3± 0 .3)× 10 6/股骨 ],P <0 .0 1;实验组BFU E和CFU GM的集落产率最低为 (11.9± 4 .6 4 ) / 3.5× 10 5BMC和 (7.9± 1.73) / 3.5× 10 5BMC ,显著高于阴性对照组 [(1.6± 1.2 ) / 3.5× 10 5BMC和 (0 .6 3± 0 .4 5 ) / 3.5× 10 5BMC],P <0 .0 0 1。结论 :“血宝”对化学药物性骨髓损伤具有保护作用 ,可明显促进骨髓损伤后造血功能的恢复     

缬沙坦涂层支架对支架术后再狭窄中胶原沉积的影响

目的评价缬沙坦涂层支架对支架置入后新生内膜中胶原沉积的影响及其预防支架内再狭窄的可行性。方法18只新西兰大白兔分为裸支架组,载体涂层支架组,缬沙坦涂层支架组3组。采用多层涂布技术制备缬沙坦涂层支架和载体涂层支架。分别将裸支架、载体涂层支架及缬沙坦涂层支架置入兔腹主动脉。术前、术后及支架置入3个月后分别行定量腹主动脉造影(QCA)测量血管直径。3个月后处死实验兔,分别测定3组支架血管段的管腔面积,内外弹力膜围绕面积,新生内膜面积及最大内膜厚度并做比较。将支架血管段进行MASSON染色,观察胶原沉积情况,天狼星红-苦味酸染色进一步观察胶原的类型。结果裸支架组(n=8),载体涂层支架组(n=8),缬沙坦涂层支架组(n=10),QCA测量的术前、术后及3个月后腹主动脉直径相似。缬沙坦组管腔面积最大,新生内膜面积最小,裸支架组、载体涂层支架组、缬沙坦涂层支架组平均管腔面积分别为(4345548±125822)μm2,(4302061±167952)μm2,(5016269±207934)μm2,平均新生内膜面积分别为(1119635±163503)μm2,(1135636±136555)μm2,(441577±74099)μm2,平均最大内膜厚度分别为(240±30)μm,(192±21)μm,(116±12)μm。MASSON染色可见新生内膜中主要是胶原沉积,天狼星红-苦味酸染色发现胶原沉积主要为Ⅲ型胶原,间或有Ⅰ型胶原。结论缬沙坦涂层支架主要是通过抑制胶原沉积抑制支架置入后血管内膜增生发挥其防治支架内再狭窄作用的。