重组2型腺相关病毒-转化生长因子β3对实验性肝纤维化大鼠肝脏组织病理学和Ⅰ型胶原表达的影响

Objective To investgate the effects of TGF β 3 on rat hepatic fibrosis. Methods The TGF β 3 cDNA was cloned into rAAV2 vector. Rats were randomly divided into four groups: normal control group, model group, negative control group and TGF β 3 group. Hepatic fibrosis was induced by hypodermic injection of 40% CCI4. Recombinant AAV2-TGF β 3 viral particles were injected via the vena caudalis one week before CCh treatment. Rats were executed 8 weeks after CCI4 treatment, global histological change was observed after HE staining, the distribution of the collagen fibers was observed after masson staining, his-tochemistry was done to observe the expression of collagen Ⅰ; The positive area rate of the collagen fibers and the average optical rate of collagen Ⅰ were quantified. Results HE staining indicated that collagen fibers were reduced in the TGF β 3 group. Masson staining shown that the collagen fibers were distributed around the blood vessel, in the portal area and disse space. Compared to the model group (13.2%±2.2%) and negative control group (12.3%±1.5%), the collagen fibers in liver tissues of TGF β3 group (7.7% ± 1.5%) were significantly decreased (q = 9.456, P < 0.01; q = 8.217, P < 0.01). Histochemistry indicated that the collagen fibers of liver tissues of TGF 15 3 group (0.185±0.033) were significantly higher than those in the model group (0.252±0.042) and the negative control group (0.230±0.029), (q = 6.228, P < 0.01; q = 4.346, P < 0.01). Conclusion TGF β 3 alleviates the damage to hepatic cell and the level offibrosis in CCI4 treated rats and inhibits the expression of collagen Ⅰ.     

重组2型腺相关病毒-转化生长因子β3对实验性肝纤维化大鼠肝脏组织病理学和Ⅰ型胶原表达的影响

目的 通过重组2型腺相关病毒(rAAV2)和基因转导,探讨转化生长因子(TGF)β 3对人鼠肝纤维化的影响. 方法构建并验证rAAV2-TGF 3.实验人鼠随机分为正常对照组、模型组、阴性对照组和TGF β 3干预组.40%的四氯化碳复制大鼠肝纤维化模型.在四氯化碳诱导前l周,将rAAV2 TGF β 3 注入大鼠尾静脉,造模8周后处死人鼠,取肝脏组织做HE染色观察肝脏组织学改变,Masson染色观察胶原纤维分布,免疫组织化学染色观察Ⅰ型胶原的表达,对胶原纤维的阳性面积比和Ⅰ型胶原的平均吸光度值进行半定量分析.多组数据问比较采用单因素的方差分析,若各组总体方差齐同,采用SNK-q检验,若方差不齐,采用Durmett'S T3检验. 结果 HE染色结果显示,rAAV2-TGF β3干预后,肝组织纤维间隔增生减少;Masson染色结果显示,人鼠胶原纤维主要分布在血管和汇管区及狄氏间隙,TGF β 3干预组大鼠肝内的胶原纤维阳性面积比为7.7%±1.5%,明显低于模型组的13.2%±2.2%和阴性对照组的12.3%±1.5%(q值分别为9.456和8.217,P值均<0.01).免疫组织化学染色结果显示,Ⅰ型胶原主要表达在血管和汇管区及狄氏间隙,TGF β 3干预组大鼠肝内的Ⅰ型胶原的表达为0.185±0.033,明显低于模型对照组的0.252±0.042和阴性对照组的0.230±0.029(口值分别为6.228和4.346,P值均<0.01).结论 TGF β 3可明显减轻实验人鼠的肝纤维化程度和组织学损伤,降低Ⅰ型胶原的表达.     

5G技术在原发性肝癌患者肝纤维化剪切波超声远程教学中的应用

目的 探讨基于5G技术的原发性肝癌患者肝纤维化超声剪切波成像技术(shear wave elastography,SWE)诊断远程教学效果。方法 选择2022年1月~2023年1月接受原发性肝癌患者肝纤维化SWE诊断教学超声专业的74名学生作为研究对象,根据班级分为甲、乙两组各37名。甲组采用传统教学模式,乙组给予基于5G技术的远程教学。观察两组学习问卷、理论知识、操作技术等得分及有效阅片量。结果 乙组学生学习问卷、理论知识、操作技术等条目得分均高于甲组,差异有统计学意义(P＜0.05)。乙组学生有效阅片量高于甲组,差异有统计学意义(P＜0.05)。结论 基于5G技术的远程教学在SWE诊断原发性肝癌患者肝纤维化的教学中显示了良好的效果。     

重组转化生长因子β3基因对大鼠肝纤维化细胞模型胶原合成及沉积的影响

目的 观察重组转化生长因子β3(TGF-β3)基因对大鼠肝纤维化细胞模型胶原合成及沉积的影响.方法 (1)构建质粒pcDNA3.1(+)-TGF-β3和pcDNA3.1(+)-TGF-β1.(2)将pcDNA3.1(+)-TGF-β1转染大鼠肝星状细胞(HSC)-T6,经G418筛选建立稳定高表达TGF-β1的HSC-T6细胞阳性克隆.(3)pcDNA3.1(+)-TGF-B3转染HSC-T6阳性细胞克隆,荧光实时定量PCR法检测TGF-β3mRNA的表达;荧光实时定量PCR法及Western印迹法分别检测TGF-β3、Ⅰ型胶原、基质金属蛋白酶(MMP)-2、MMP-9及TIMP-1的mRNA和蛋白表达情况.结果 (1)pcDNA3.1(+)-TGF-β3和pcDNA3.1(+)-TGF-β3质粒均可转染HSC-T6细胞,以转染48 h时转染效率最高,达28.2%.(2)pcDNA3.1(+)-TGF-β3转染HSC-T6细胞阳性克隆后,与单纯阳性克隆组相比,TGF-β1mRNA表达无明显改变,而蛋白表达明显低(0.48±0.07 vs 0.66±0.14,P=0.023);Ⅰ型胶原mRNA及蛋白的表达明显低(mRNA:7.2±1.0 vs 13.7±0.4,P=0.010,蛋白:0.45±0.21 vs 0.82±0.13,P=0.041),MMP-2 mRNA及蛋白较低,但差异无统计学意义(均P>0.05),MMP-9 mRNA及蛋白表达明显多(均P<0.05),TIMP-1 mRNA及蛋白表达明显低(均P<0.05).结论 (1)重组TGF-β3真核表达载体构建正确,在转染阳性克隆细胞48 h后获得高效表达;(2)稳定高表达TGF-B1的HSC-T6细胞阳性克隆可作为肝纤维化细胞模型;(3)pcDNA3.1(+)-TGF-β3转染阳性克隆能减少胶原合成,并通过调节基质金属蛋白酶及其抑制剂的表达对胶原沉积起抑制作用.     

重组2型腺相关病毒-转化生长因子β3对实验性肝纤维化大鼠肝脏组织病理学和Ⅰ型胶原表达的影响

Objective To investgate the effects of TGF β 3 on rat hepatic fibrosis. Methods The TGF β 3 cDNA was cloned into rAAV2 vector. Rats were randomly divided into four groups: normal control group, model group, negative control group and TGF β 3 group. Hepatic fibrosis was induced by hypodermic injection of 40% CCI4. Recombinant AAV2-TGF β 3 viral particles were injected via the vena caudalis one week before CCh treatment. Rats were executed 8 weeks after CCI4 treatment, global histological change was observed after HE staining, the distribution of the collagen fibers was observed after masson staining, his-tochemistry was done to observe the expression of collagen Ⅰ; The positive area rate of the collagen fibers and the average optical rate of collagen Ⅰ were quantified. Results HE staining indicated that collagen fibers were reduced in the TGF β 3 group. Masson staining shown that the collagen fibers were distributed around the blood vessel, in the portal area and disse space. Compared to the model group (13.2%±2.2%) and negative control group (12.3%±1.5%), the collagen fibers in liver tissues of TGF β3 group (7.7% ± 1.5%) were significantly decreased (q = 9.456, P < 0.01; q = 8.217, P < 0.01). Histochemistry indicated that the collagen fibers of liver tissues of TGF 15 3 group (0.185±0.033) were significantly higher than those in the model group (0.252±0.042) and the negative control group (0.230±0.029), (q = 6.228, P < 0.01; q = 4.346, P < 0.01). Conclusion TGF β 3 alleviates the damage to hepatic cell and the level offibrosis in CCI4 treated rats and inhibits the expression of collagen Ⅰ.     

TGF-β家族在肝纤维化中的不同作用及对策

转化生长因子β(TGF-β)是一组多功能多肽生长因子,参与细胞生长分化、免疫调节、伤口愈合、血管形成、胚胎形成和细胞凋亡等．迄今为止,哺乳动物类发现有3种TGF-β,即TGF-β1,TGF-β2和TGF-β3．尽管在许多实验中3种TGF-β生物行为相似,但是在胚胎形成、组织纤维化及伤口愈合过程中他们的调控机制及表达各不相同,尤其在肝纤维化形成过程中起着不同的作用,本文就TGF-β3种亚型与肝纤维化之间关系的最新研究进展作一综述．     

重组转化生长因子-β3基因对大鼠肝星状细胞内、外蛋白表达的影响

目的探讨重组转化生长因子-β3（TGF-β3）基因对大鼠肝星状细胞（HSC）内、外蛋白合成及分泌的影响。方法构建质粒pcDNA3.1（＋）-TGF-β3和pcDNA3.1（＋）-TGF-β1。稳定转染：通过脂质体介导的方法,将pcDNA3.1（＋）-TGF-β1转染HSC-T6,经G418筛选建立稳定高表达pcDNA3.1（＋）-TGF-β1的HSC-T6细胞阳性克隆。再将pcDNA3.1（＋）-TGF-β3转染HSC-T6克隆细胞,用W estern b lot法和酶联免疫吸附法（ELISA）分别检测细胞内外TGF-β1、Ⅰ型胶原、基质金属蛋白酶（MMP）-2、MMP-9及金属蛋白酶组织抑制剂（TIMP）-1的蛋白表达量。结果pcDNA3.1（＋）-TGF-β3转染HSC-T6阳性细胞克隆后,与单纯阳性克隆组相比,TGF-β1、Ⅰ型胶原、TIMP-1细胞内蛋白表达及培养上清中的分泌水平均明显降低（P〈0.05）,MMP-2的表达也降低,但两者间差异无统计学意义（P〉0.05）,而MMP-9的表达明显增高（P〈0.05）。结论重组质粒pcDNA3.1（＋）-TGF-β3能减少细胞内外Ⅰ型胶原的合成及分泌;通过调节MMP-9及TIMP-1的表达,可抑制细胞外基质的合成,促进其降解。     

对117例老年功能性便秘的综合护理干预观察

探索综合护理干预对老年功能性便秘患者的功效。117例入院前即有功能性便秘的老年患者给予综合护理干预后,随访观察4周,评估便秘改善情况。综合护理干预措施包括向患者及家属讲解便秘的危害,改正不良饮食习惯,定时排便,每天按摩腹部及适量运动,针对卧床排便患者提供舒适的环境及姿势、药物等。 综合护理干预使部分便秘患者的病症程度减轻。 采取个性化的综合护理干预能有效改善老年功能性便秘患者的便秘情况。     

普通IFNα-2b治疗慢性乙型肝炎患者外周血中性粒细胞和血小板计数的变化

目的进一步认识普通IFNα-2b治疗慢性乙型肝炎患者外周血中性粒细胞和血小板的变化规律。方法接受普通IFNα-2b治疗的24例CHB病人,观察48周。结果干扰素治疗后中性粒细胞数减少在第2周最明显,血小板减少在第4周内最明显,以后均逐渐恢复。结论干扰素所致外周血中性粒细胞和血小板数减少是可逆性的。