福辛普利对大鼠肾小球系膜细胞转化生长因子-β1蛋白及其mRNA表达的影响

目的观察血管紧张素酶抑制剂（ACEI）-福辛普利（FOS）对脂多糖（LPS）诱导大鼠肾小球系膜细胞（GMC）转化生长因子-β（TGF-β1）蛋白及mRNA表达的影响，探讨FOS延缓肾小球硬化机制。方法按经典方法体外培养大鼠GMC，转种培养后分为对照组、LPS组、LPS＋FOS组。采用ELISA法测定GMC培养上清6、12、24hTGF-β1蛋白水平，荧光半定量RT-PCR法检测TGF-β1mRNA表达。结果LPS诱导组GMC分泌TGF-β1蛋白及mRNA表达均高于对照组，LPS＋FOS组GMC分泌TGF-β1及mRNA表达较LPS组明显下降。结论FOS从蛋白和mRNA两个水平均对体外培养大鼠GMC产生TGF-β1有明显抑制作用，提示FOS抑制GMC产生TGF-β1可能是延缓肾小球硬化的重要作用机制之一。     

转化生长因子β1在大鼠心肌缺氧缺血再灌注损伤修复及心肌重塑中的作用

目的 探讨内源性转化生长因子β1（TGF-β1）在心肌缺氧缺血-再灌注损伤修复及心肌重塑中的作用。方法 根据新生鼠常压窒息模型，制作缺氧缺血-再灌注损伤大鼠动物模型，将60只新生大鼠随机分成缺氧缺血-再灌注损伤1d组（A组）、7d组（B组）、14d组（C组）以及正常对照组（D组）4组。每组15只用免疫组化方法对TGF-β1在心肌的分布及表达进行定性及半定量分析，用Masson染色方法测定心肌胶原容积（CVF），同时利用HE染色观察心肌组织的病理学改变。统计分析应用SPSS11.0统计软件进行。多组均数比较采用单因素方差分析，两变量之间的关系采用相关分析。结果 TGF-β1表达：B组及C组分别与对照组相比，均有统计学意义（P均〈0．01），胶原含量（CVF）：B组及C组较正常对照组差异有统计学意义（P均〈0．01），相关分析表明，CVF与TGF-β1之间呈正相关（r=0．574，P〈0．01）。结论 大鼠缺氧缺血-再灌注损伤后TGF-β1表达量随损伤时间不同而变化，其可能参与缺氧缺血-再灌注所致的内脏损伤后的自我修复。缺氧缺血-再灌注损伤后TGF-β1的表达与心肌胶原含量的变化呈正相关，提示TGF-β1可能参与损伤后心肌重塑。     

HIF-1α特异性RNA干扰表达载体的构建及体外效应研究

目的：己知缺氧诱导因子-1α（HIF-1α）在缺氧状态下可通过调节内皮素-1（ET-1）的升高而参与肺血管内皮细胞损伤及肺动脉高压,最终导致肺出血。RNA干扰(RNAi)是双链RNA介导的序列特异性转录后同源靶基因沉默效应,目前RNAi技术已成为一种研究基因功能的有力工具,故拟通过构建人HIF-1α基因的特异性小干扰RNA（siRNA）真核表达载体,观测该载体在缺氧状态下对HIF-1α基因的干扰作用及对ET-1的抑制作用。方法：:选择6个（a～f）可能的人HIF-1α siRNA干扰位点,设计合成相应的特异性互补寡聚核苷酸链（A～F）,采用基因克隆技术,将合成的（A～F）链序列定向插入真核表达载体中,构建成重组质粒HIF-1α siRNA真核表达载体（A′～F′）,通过脂质体法转染至体外人脐静脉内皮细胞（HUVECs）48 h,再以CoCl2（100μM）模拟缺氧刺激3 h后,观测siRNA对HIF-1α mRNA和HIF-1 α蛋白表达的抑制效应及ET-1水平。结果：①成功构建了分别针对6个HIF-1 α siRNA干扰位点（a～f）的重组质粒HIF-1α siRNA真核表达载体（A′～F′）;②该6组真核表达载体转染HUVECs并经CoCl2模拟缺氧刺激后,与未转染组比较,结果显示B′ 及D′ 组明显抑制HIF-1α mRNA及HIF-1α蛋白的表达,并部分抑制ET-1表达,而其余4组与未转染组比较并无差异。结论：针对b及d干扰位点的B′及D′特异性siRNA真核表达载体,能明显干扰HIF-1α基因的表达,进而抑制ET-1的释放。该实验为下一步用B′、D′特异性siRNA真核表达载体在动物体内研究HIF-1α与新生儿缺氧性肺出血关系奠定基础。     

转化生长因子β1在大鼠窒息复氧心肌损伤后心肌重塑中的作用

目的观察大鼠窒息复氧心肌损伤后心肌内源性转化生长因子（TGF）-β1表达，探讨TGF-β1在心肌缺氧缺血再灌注损伤修复及心肌重塑中的作用。方法制作窒息复氧心肌损伤大鼠动物模型，将60只新生大鼠随机分成窒息复氧心肌损伤1d组（A组）、7d组（B组）、14d组（C组）以及正常对照组（D组）。每组15只，快速定量检测血清心肌肌钙蛋白（cTnI）水平，心肌TGF-β1表达采用免疫组化方法进行定性及半定量分析，用Masson染色方法测定心肌总胶原容积（CVF），同时利用HE染色观察心肌组织的病理学改变。结果血清cTnI呈一过性增高，A组明显高于D组，差异非常显著（P〈0．01），B、C组下降，与D组比较差异无显著性（P〉0．05）。TGF-β1表达及CVF随复氧时间延长而增加，A组轻度增高，但较D组差异无显著性（P〉0．05），B组明显增高，C组达高峰，与D组比较差异均非常显著（P〈0．01）。CVF与TGF-β1表达呈正相关（r=0．574，P〈0．01）。结论大鼠窒息复氧心肌损伤后，TGF-β1表达随复氧时间的不同而变化，可能参与了心肌损伤后的自我修复及心肌重塑。     

氯沙坦片对糖尿病肾病大鼠转化生长因子-β1及结缔组织生长因子的影响

目的 观察糖尿病肾病（DN）大鼠转化生长因子-β1（TGF-β1）、结缔组织生长因子（CTGF）在肾组织中的表达，检测尿液中两者的水平。观察氯沙坦片对肾组织TGF-β1、CTGF蛋白的表达及尿液中水平的影响。方法 Wistar大鼠摘除右肾后，注射链脲菌素制造DN大鼠模型。随机分为糖尿病肾病实验组和氯沙坦治疗组。于第6、12周用免疫组织化学法检测肾组织TGF-β1、CTGF蛋白质的表达，ELISA法检测尿液中两者水平。结果 与治疗组相比，实验组12周时尿液TGF-β1、CTGF水平、肾间质纤维组织相对面积显著增加，肾组织TGF-β1、CTGF蛋白质的表达显著升高（P均〈0．01）；肾小球系膜基底膜相对面积扩大（P〈0．05）。与6周相比，实验组12周时尿液TGF-β1、CTGF水平，肾间质纤维组织相对面积显著增加。肾组织内TGF-β1、CTGF蛋白质的表达亦明显升高（P均〈0．01）；治疗组12周肾小球TGF-β1蛋白质的表达升高（P〈0．05），肾小管TGF-β1蛋白质的表达明显升高（P〈0．01）。肾组织内TGF-β1蛋白与CTGF蛋白的表达呈显著正相关（r=0．85P〈0．01），尿液中两者排泄量亦呈明显正相关（r=0．76P〈0．01）。结论 TGF-β1、CTGF蛋白在DN大鼠肾组织表达及尿液中的排泄显著升高。氯沙坦可减轻肾组织TGF-β1、CTGF蛋白的表达，减少两者尿液中的排泄，具有肾保护作用。     

microRNA-145对TGF-β1诱导的人肾小管上皮细胞上皮间充质转化的影响

目的研究micro RNA-145(miR-145)对转化生长因子(TGF)-β1诱导人肾小管上皮细胞(HK-2)上皮间充质转化(EMT)的影响。方法人工合成miR-145基因序列,构建真核重组质粒p CMVmiR-145。以未处理HK-2细胞为对照组;以TGF-β1处理HK-2细胞为TGF-β1组;以p CMV-myc空白质粒转染后经TGF-β1处理HK-2细胞为TGF-β1空白质粒组;以p CMV-miR-145质粒转染后经TGF-β1处理HK-2细胞为TGF-β1+miR-145组。采用实时荧光定量PCR检测miR-145表达。采用Western blot检测TGF-β/Smad信号传导蛋白TGF-β1、Smad3、Smad2/3、p-Smad2/3,以及EMT生物标记物蛋白α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)和I型胶原蛋白(ColⅠ)的表达水平。采用ELISA法检测培养细胞上清液中FN和Col I的含量。结果miR-145表达质粒构建成功,重组质粒有效转染TGF-β1诱导HK-2细胞。与对照组比较,TGF-β1+miR-145组细胞中miR-145表达上调(P0.01);与对照组和TGF-β1+miR-145组比较,TGF-β1组和TGF-β1空白质粒组细胞中miR-145表达均下降(P0.01)。与TGF-β1组和TGF-β1空白质粒组比较,TGF-β1+miR-145组细胞内TGF-β1、Smad3、Smad2/3和p-Smad2/3蛋白表达量减少(P0.05);α-SMA、FN、ColⅠ蛋白表达亦明显减少(P0.05);TGF-β1+miR-145组培养液上清中FN、ColⅠ含量减少(P0.05)。结论 miR-145参与TGF-β1处理HK-2细胞EMT的调控。miR-145可能通过抑制TGF-β依赖的Smad信号通路活性,从而抑制肾小管EMT。     

Apollon siRNA联合川芎嗪对白血病细胞增殖及凋亡的影响

目的 观察Apollon siRNA 靶向沉默Apollon 基因联合川芎嗪（TMP）对人慢性粒细胞白血病细胞株K562 增殖及凋亡的影响。方法 根据前期实验筛选出的干扰效果最佳的Apollon siRNA 片段,构建pGPHIGFP-Neo-Apollon 真核表达载体,并将构建成功的载体转染至K562 细胞。将实验分为细胞对照组（未行任何处理）、阴性对照组（转染阴性质粒载体）和RNA 干扰组（转染pGPHI-GFP-Neo-Apollon 质粒载体）,利用RT-PCR 法和细胞免疫荧光法分别检测各组细胞Apollon mRNA 及蛋白的表达情况;再于上述分组基础上新增TMP 组（施加320 μg/mL TMP）、TMP+ 阴性对照组和TMP+RNA 干扰组,应用MTT 法和流式细胞术分别检测各组K562 细胞的增殖能力和细胞凋亡率。结果 构建的pGPHI-GFP-Neo-Apollon 载体能在K562 细胞内稳定表达;RNA 干扰组Apollon mRNA 及其蛋白的相对表达水平明显低于细胞对照组和阴性对照组（均P<0.05）;RNA 干扰组K562细胞的增殖抑制率和凋亡率高于细胞对照组（P<0.05）,与RNA 干扰组及TMP 组比较,siRNA 转染联合TMP能显著提高K562 细胞的增殖抑制率和凋亡率（均P<0.05）。结论 Apollon siRNA 转染能显著抑制K562 细胞增殖并促进其凋亡,且与TMP 联合使用对提高K562 细胞的增殖抑制率和凋亡率具有显著协同作用,提示siRNA技术联合药物在白血病治疗中具有重要的潜在价值。     

罗格列酮对大鼠系膜增生性肾炎的治疗作用

目的 研究罗格列酮对大鼠系膜增生性肾炎(MsPGN)的治疗作用并探讨其可能的作用机制,寻找治疗系膜增生性肾炎的新途径.方法 大鼠随机分为3组:李白对照组(A组)、模型对照组(B组)、罗格列酮治疗组(C组).B、C组以四氯化碳及牛血清白蛋白灌胃制备系膜增生性肾炎模型,制模成功后.A、B组予生理盐水,治疗组予罗格列酮分别灌胃均8周,每周测各组大鼠24 h尿蛋白排泄量及尿红细胞计数.实验结束时,测血清白蛋白、血糖、肝肾功能、肾脏系膜细胞及系膜基质增生情况(光镜、PAS染色)及各组大鼠肾脏转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)表达水平.结果 治疗组大鼠24 h尿蛋白排泄量、尿红细胞汁数、TGF-β1及α-SMA表达水平、肾小球PAS染色灰度值较模型组均明显减少,血清白蛋白明显升高,差异均有统计学意义.3组之间血精及肝肾功能比较差异无统计学意义.结论 罗格列酮对大鼠系膜增生性肾炎具有治疗作用,其作用机制之一可能是通过抑制TGF-β1及其下游因子表达.     

应用RNA干扰技术抑制骨肉瘤细胞中基质金属蛋白酶-1基因表达的研究

目的构建针对人基质金属蛋白酶-1（MMP-1）基因的特异性短发卡RNA（shRNA）真核表达载体，并观察其在人骨肉瘤细胞MG63中对MMP-1基因表达的抑制作用。方法培养骨肉瘤细胞MG63，根据MMP-1基因cDNA编码序列，设计并合成针对MMP-1基因的特异性RNA干扰片断，将其克隆入pSilencer 3．0-H1 neo质粒，构建MMP-1基因shRNA真核表达载体siRNA MMP-1。与阴性对照质粒分别转染MG63细胞，G418筛选稳定转染的细胞系。采用RT—PCR、Western blot检测MMP-1 mRNA和蛋白的表达水平，同时进行体外侵袭试验。结果成功构建了MMP-1基因shRNA真核表达载体siRNA MMP-1。获得了稳定转染siRNA MMP-1载体和阴性对照载体siRNA neg的细胞系，并发现MG63／siRNA MMP-1细胞MMP-1 mRNA和蛋白表达均受到明显抑制，细胞侵袭能力也显著下降。结论特异性shRNA能够明显抑制MMP-1基因在MG63细胞中的表达，这为进一步研究MMP-1在MG63细胞中的生物学功能和作用机制奠定了实验基础。     

UCP2 基因siRNA 转染对脓毒症血清诱导心肌细胞炎症反应的影响

目的 研究解偶联蛋白2 基因（uncoupling protein 2, UCP2）siRNA 干扰大鼠H9C2 心肌细胞株后对脓毒症血清诱导炎症反应的影响,探讨UCP2 在脓毒症心肌病可能的机制。方法 制备正常大鼠血清和脓毒症大鼠血清。体外培养大鼠心肌细胞株（H9C2）,随机分为空白对照组、正常大鼠血清、10%脓毒症大鼠血清刺激12 h 组（脓毒症血清组）、UCP2-siRNA 干扰+10%脓毒症大鼠血清刺激12 h 组（UCP2-siRNA+ 脓毒症血清组）、阴性siRNA 转染+10%脓毒症大鼠血清刺激12 h 组（阴性siRNA+ 脓毒症血清组）。RT-PCR 检测各组TNF-α mRNA,IL-1βmRNA 表达;免疫印迹法检测磷酸化p38MAPK（p-p38MAPK）表达和核转录因子（NF-κB）的表达。结果 UCP2-siRNA+ 脓毒症血清组 p-p38、NF-κB 表达量均较脓毒症血清组明显增高（PP结论 UCP2 参与脓毒症血清诱导的H9C2 心肌细胞p38MAPK 活性和NF-κB 与下游炎症介质的表达调控。     

生物标记物粪便胆汁酸的测定在过敏性紫癜患儿诊治中的意义

目的 探讨生物标记物粪便胆汁酸浓度在过敏性紫癜(HSP)患者中的变化及其在诊治中的临床意义。方法 选取2014~2016年确诊为HSP的19例患儿为HSP组,另选取27例健康儿童为健康对照组。采集HSP组患儿急性期、恢复期及健康对照组儿童粪便标本,应用液相质谱技术检测各组儿童粪便胆汁酸水平。结果 HSP组患儿恢复期胆酸水平均高于健康对照组和HSP组急性期 (P < 0.016)。HSP组患儿恢复期鹅脱氧胆酸水平高于健康对照组 (P < 0.016)。HSP组患儿急性期和恢复期脱氧胆酸、石胆酸水平均低于健康对照组 (分别P < 0.05、P < 0.016)。各组间熊去氧胆酸水平比较差异均无统计学意义 (P > 0.05)。结论 HSP患儿急性期粪便次级胆汁酸脱氧胆酸和石胆酸低于健康对照组,这可能与HSP的发病或转归有关。     

霉酚酸酯和环磷酰胺治疗儿童大量蛋白尿型过敏性紫癜性肾炎的前瞻性随机对照研究

目的 探讨霉酚酸酯(MMF)和环磷酰胺(CTX)分别治疗具有大量蛋白尿[24?h尿蛋白定量≥50?mg/kg或晨尿蛋白/肌酐(mg/mg)≥2.0]的过敏性紫癜性肾炎(HSPN)患儿的疗效和安全性.方法 前瞻性纳入2016年8月至2019年11月在首都儿科研究所附属儿童医院肾脏内科住院并诊断为有大量蛋白尿的HSPN患儿...     

EFFECT OF CYPROTERONE ACETATE (CA) ON GROWTH AND ENDOCRINE FUNCTION IN PRECOCIOUS PUBERTY

Abstract. Stahnke, N., Ilicki, A. and Willig, R. P. (Department of Paediatrics, University Hospital, Hamburg, West Germany). Effect of cyproterone acetate (CA) on growth and endocrine function in precocious puberty. Acta Paediatr Scand, Suppl. 277: 32, 1979.–16 girls with precocious puberty have been studied. Following low dosage cyproterone acetate (CA) therapy (mean daily dosage 65 mg/m² BSA) a beneficial effect on growth and skeletal maturation was observed. During high dosage therapy (150 mg/m² per day) endocrinological studies were performed in 10 of these patients. There was no significant difference in HGH levels (insulin-and arginine-test), T₃ and TSH values (TRH-test) between patients and controls, T₄ concentration was significantly increased. Basal prolactin levels and prolactin response to TRH was definitely elevated. Oral glucose load and arginine infusion resulted in a significantly enhanced insulin release. There was a significant reduction in basal LH levels and an increase in FSH response to LH-RH. Basal and diurnal plasma cortisol values were markedly reduced and the cortisol release due to corticotrophin injection, (lsinevasopressin (LVP) injection and insulin-hypoglycemia as well. A definite increase in basal ACTH levels was observed, during LVP-and insulin-hypoglycemia test ACTH concentrations were within or significantly above normal range. In our patients a primary adrenocortical insufficiency due to CA treatment was evident.     

Obesogenic home food availability,diet, and BMI in Pakistani and White toddlers

Individuals of South Asian ethnicity have an increased risk for obesity and related diseases. Foods available in the home during the first 1000 days (conception to 24 months old) are an important determinant of diet, yet no study has examined the association of early‐life home food availability (HFA) with later diet and obesity risk in South Asian households. We examined whether obesogenic HFA at 18 months of age is associated with dietary intake and body mass index (BMI) at 36 months of age in low‐income Pakistani and White households in the United Kingdom. In this prospective birth cohort study (Born in Bradford 1000), follow‐up assessments occurred at 18 (n = 1032) and 36 (n = 986) months of age. Variety and quantity of snack foods and sugar‐sweetened beverages (SSBs) in the home and consumed were measured using the HFA Inventory Checklist and food frequency questionnaires, respectively. BMI was calculated using measured length/height and weight. Multinomial logistic regression models examined associations between HFA and tertiles of dietary intake, and multivariable linear regression models assessed associations between HFA and BMI. Pakistani households had a greater variety and quantity of snack foods and SSBs available compared with White households. Variety and quantity of snack foods and SSBs in the home at 18 months were positively associated with children''s intake of these items at 36 months, but associations between HFA and BMI were null. Reducing obesogenic HFA during the first 1000 days may promote the development of more healthful diets, though this may not be associated with lower obesity risk during toddlerhood.     

Prenatal dietary diversity may influence underweight in infants in a Ugandan birth‐cohort

Growth faltering in early childhood is prevalent in many low resource countries. Poor maternal dietary diversity during pregnancy has been linked with increased risk of fetal growth failure and adverse birth outcomes but may also influence subsequent infant growth. Our aim is to assess the role of prenatal maternal dietary diversity in infant growth in rural Uganda. Data from 3291 women and infant pairs enrolled in a birth cohort from 2014 to 2016 were analysed ({"type":"clinical-trial","attrs":{"text":"NCT04233944","term_id":"NCT04233944"}}NCT04233944). Maternal diets were assessed using dietary recall in the second or third trimesters of pregnancy. Maternal dietary diversity scores (DDS) were calculated using the FAO Minimum Dietary Diversity for Women (MDD‐W). Cox regression models were used to evaluate associations of the DDS with the incidence of underweight, stunting and wasting in infants from 3 to 12 months, adjusting for confounding factors. The median DDS for women was low, at 3.0 (interquartile range 3.0–4.0), relative to the threshold of consuming five or more food groups daily. Infants of women in highest quartile of DDS (diverse diets) were less likely to be underweight (adjusted hazard ratio: 0.70, 95% confidence interval: 0.61, 0.80) compared with infants of women in Quartile 1 (p for trend <0.001) in models controlling for maternal factors. There was no significant association between DDS and stunting or wasting. Our findings suggest a relationship between higher maternal dietary diversity and lower risk of underweight in infancy. These findings suggest that programmes to improve infant growth could additionally consider strengthening prenatal dietary diversity to improve child outcomes globally.     

Blurring the distinctions between on and off the job injuries: similarities and differences in circumstances.

OBJECTIVES: To compare the causes of non-fatal work and non-work injuries and the places or environments where they occur. It has been suggested that many injuries may have similar etiologies on and off the job and thus involve some common prevention strategies. However lack of comparable data on work relatedness has prevented testing this proposition. METHODS: The National Health Interview Survey (NHIS) now collects information on the cause, location, and work relatedness of all medically attended injuries. National US estimates of non-fatal work and non-work injuries were compared by cause and place/location for working age adults (18-64 years). RESULTS: Overall 28.6% of injuries to working age adults were work related (37.5% among employed people). The causes and locations of many work and non-work injuries were similar. Falls, overexertion, and struck/caught by were leading causes for work and non-work injuries. Motor vehicle injuries were less likely to be work related (3.4% at work v 19.5% non-work) and overexertion injuries more likely to be work related (27.1% v 13.8%). Assaults were less than 1% of work injuries and 1.8% of non-work injuries. Both work and non-work injuries occurred in every location examined-including the home where 3.5% of injuries were work related. CONCLUSIONS: Work and non-work injuries share many similarities suggesting opportunities to broaden injury prevention programs commonly restricted to one setting or the other. Comprehensive efforts to prevent both non-work and work injuries may result in considerable cost savings not only to society but also directly to employers, who incur much of the associated costs.