目的:探讨护患互动达标模式干预对妊娠期糖尿病(GDM)患者自护能力、血脂、血糖及妊娠结局的影响。方法:选取上海交通大学医学院附属国际和平妇幼保健院产科2018年5月至2019年2月收治的GDM患者100例，随机分为对照组和试验组，每组50例。对照组患者进行常规护理干预，试验组患者采用护患互动达标模式进行护理干预，比较两组患者自护能力、血脂、血糖水平和妊娠结局的差异。结果:干预前两组患者血糖水平比较差异无统计学意义( P>0.05)；干预后2 w，试验组患者的空腹血糖(FPG)、餐后2 h血糖(2 hPG)和糖化血红蛋白(HbA1c)水平均明显低于对照组，差异有统计学意义( P<0.05)。干预前两组患者自护能力评分差异无统计学意义( P>0.05)；干预后2 w，试验组患者自护能力各维度评分均明显高于对照组，差异有统计学意义( P<0.05)。试验组中孕妇不良结局发生率和胎儿不良结局发生率明显低于对照组，差异有统计学意义( P<0.05)。 结论:运用护患互动达标模式对GDM患者进行护理干预，可有效提高患者的疾病自护能力，改善其血糖控制水平，从而减少不良妊娠结局的发生。

目的 探讨长链非编码RNA(long noncoding RNA,lncRNA)-AF289591对间充质干细胞(hu-man mesenchymal stem cells,hMSC)成骨分化的影响及其分子机制.方法 收集骨质疏松症(osteoporosis,OP)患者以及健康人群的骨髓标本,通过lncRNA芯片分析hMSC中lncRNAs的表达,并通过实时荧光定量聚合酶链式反应(real-time polymerase chain reaction,qPCR)验证lncRNA-AF289591在两类人群中的表达,ELISA检测骨髓组织中硬骨抑素(sclerostin)水平.分离hMSC并通过流式细胞技术进行细胞鉴定,qPCR检测hMSC骨形成相关基因COL、OPN、OCN表达以及SOST基因表达水平,试剂盒检测碱性磷酸酶(alkaline phosphatase,ALP)活性,免疫印迹法(Western blot)检测硬骨抑素和RUNX2蛋白表达水平,荧光素酶报告基因检测Wnt-1启动子活性.结果 相比健康人群,OP患者骨髓组织中lncRNA-AF289591表达显著升高,伴随着SOST基因表达水平增加(P<0.05).分离hMSC并通过流式细胞技术确定为阳性细胞(CD105、CD44和CD29表达百分率分别为99.9％、100％和99.8％),过表达lncRNA-AF289591后,抑制骨形成相关分子COL、OPN、OCN基因表达(P<0.05),降低ALP活性(P<0.05),促进SOST基因表达(P<0.05),抑制Wnt-1启动子活性以及减少RUNX2蛋白表达水平(P<0.05).干扰lncRNA-AF289591表达后,结果与上述情况相反(P<0.05).结论 lncRNA-AF289591表达通过调控SOST基因表达而抑制hMSC成骨分化.

目的 分析X连锁显性低血磷性佝偻病(X-linked dominant hypophosphatemia,XLH)患者的临床表型、 生化特点及X染色体上内肽酶同源磷调节基因(phosphate-regulating gene with homologies to endopepti-dase on the X chromosome,PHEX)突变特征.方法 收集2016至2018年就诊于上海交通大学附属第六人民医院骨质疏松和骨病专科的19例XLH患者的临床资料,利用sanger测序技术进行PHEX基因突变检测,酶联免疫吸附试验检测血清成纤维生长因子23(fibroblast growth factor 23,FGF23)水平.结果 确诊19例PHEX突变导致XLH患者,其中男性7例,女性12例;9例为散发;共鉴定出13个不同PHEX突变位点,5个新发突变.未成年患者身高均低于同年龄组,4例低于同年龄组1个标准差,4例低于同年龄组2个标准差.成年男性患者和女性患者身高分别为130 cm(n=1)和142(134～146)cm(n=6).不同程度的骨痛5/11例,牙齿疾患10/11例,下肢弯曲畸形16/16例.XLH患者血清全段FGF23水平[80.8(45.0～141.9)pg/mL]存在不同程度升高,与正常对照组[40.6(33.9～51.8)pg/mL]比较,差异有统计学意义(P=0.002).结论 本研究丰富了PHEX基因致病的表型谱及突变谱;散发病例高发,应避免散发患者的漏诊、 误诊;需重视基因鉴定及产前诊断对该病预防及诊治的重要性.

目的 从细胞水平探索氯沙坦钾对骨骼肌损伤的保护作用及其机制.方法 用2％马血清诱导分化L6成肌细胞;CCK-8试剂盒及Western Blot确定合适的氯沙坦细胞处理浓度.实验分为4组:对照组,二甲基亚砜组,棕榈酸(1 mmol/L)组以及棕榈酸(1 mmol/L)+氯沙坦钾(50μmol/L)组.Western Blot检测凋亡相关蛋白(cleaved caspase-3、cleaved PARP、Bcl-2、Bax)、自噬相关蛋白(LC3、Beclin 1),实时荧光定量PCR(quantitative real-time PCR,qPCR)检测P53和P21 mRNA表达水平.结果 显微镜下观察及Western Blot检测myogenin蛋白表达增加,证明骨骼肌细胞诱导分化成功.CCK-8试剂盒检测示氯沙坦钾浓度为50、100、200、400μmol/L时L6成肌细胞活力分别为0.87±0.087、1.11±0.037、1.21±0.099、1.30±0.068,其中50μmol/L时细胞活力最大,且对cleaved caspase-3蛋白表达抑制最明显.与未作处理的对照组比较,qPCR显示棕榈酸处理组骨骼肌细胞P53、P21 mRNA表达量分别从0.36±0.03、0.50±0.05上调至0.68±0.08、1.07±0.02(P<0.05),cleaved caspase3、cleaved PARP蛋白表达均增加,LC3Ⅱ/LC3Ⅰ及Beclin 1蛋白表达减低;与棕榈酸单独处理组比较,氯沙坦钾与棕榈酸共处理组P53、P21 mRNA表达量分别从0.68±0.08、1.07±0.02下调至0.44±0.01、0.67±0.03(P<0.05),cleaved caspase-3、cleaved PARP蛋白表达减低,LC3Ⅱ/LC3Ⅰ及Beclin1蛋白无显著改变.结论 氯沙坦钾通过抑制cleaved caspase3及其底物cleaved PARP表达抑制细胞凋亡,但这种保护效应并不是通过自噬途径实现的.

A system of offline hydrophilic/reversed-phase two-dimensional liquid chromatography coupled with mass spectrometry (HILIC-RPLC-MS) was developed to analyze phospholipid in serum samples.A HILIC column was used in the first dimensional chromatography with gradient elution using ammonium acetate as a buffer and acetonitrile as mobile phase.A C18 column was used in the second dimensional chromatography with gradient elution using ammonium formate as a buffer and methanol as mobile phase.The effluent was detected by mass spectrometry with electrospray ionization(ESI) under positive and negative scan modes.Results showed that,compared with 1D LC-MS system,the 2D LC-MS provided higher peak capacity and better separation efficiency,especially for low abundance phospholipid.And this method reduced coelution and ion suppression effect.It can be used to separate and analyze serum and other complex biological systems,and lays a foundation for phospholipidomic research.

目的 研究白藜芦醇(RSV)对脂多糖(LPS)诱导的H9c2细胞损伤的保护作用及其机制.方法 用LPS 10 μg/mL处理大鼠H9c2心肌细胞0、5、10、20、30、60 min,评估LPS在不同时间点对H9c2细胞核因子κB抑制蛋白(IκBα)、p-p65、p65和p-肌动蛋白(β-actin)表达的诱导作用;用不同浓度RSV预处理H9c2细胞后再用LPS刺激,观察RSV对LPS诱导的H9c2细胞损伤的影响.MTT法检测H9c2细胞增殖能力;荧光法检测细胞内活性氧(ROS)水平;Western blotting检测IκBα、p-p65、p65和β-actin的表达.结果 LPS抑制H9c2细胞增殖,使细胞内ROS产生量明显增加;LPS使IκBα表达水平显著降低,使p65磷酸化水平显著升高.RSV预处理能明显降低LPS对H9c2细胞的损伤,使细胞内ROS产生明显减少;RSV可上调IκBα表达,下调p-p65的表达.结论 RSV可能通过下调p-p65及上调IκBα抑制LPS诱导的H9c2细胞损伤.

目的:评价中西医结合方法在改善OME抑制及分解MUC黏蛋白的作用.方法:SD大鼠50只,随机分为五组,四组实验组(模型组、中药组、西药组、中西医组),建立肺炎链球菌大鼠分泌性中耳炎模型,采用RT-PCR技术检测MUC2、MUC5A mRNA在各组中耳黏膜中的表达.结果:与正常组比较,模型组MUC2、MUC5A含量显著增高;与模型组比较,各药物组合量显著下降,中西医组下降更为明显.结论:加味二陈汤联合厄多司坦可以更好地改善大鼠中耳黏膜蛋白MUC2、MUC5A分泌情况.

Objective To investigate whether resveratrol ( RSV) protects H9c2 cells against lipopolysaccharide ( LPS) induced oxi-dative injury partly through ERK1/2 signaling pathway.Methods H9c2 cells were divided into six groups:control, LPS (10μg/ml li-popolysaccharide),L+R5(10μg/ml lipopolysaccharide +5 μmol/L resveratrol),L+R10(10μg/ml lipopolysaccharide +10μmol/L res-veratrol),L+R20(10μg/ml lipopolysaccharide +20 μmol/L resveratrol),L +R50(10μg/ml lipopolysaccharide +50μmol/L resvera-trol).H9c2 cells from group L+R5 ,L+R10,L+R20,L+R50 were pretreated with resveratrol .Then, cells from group LPS ,L+R5,L+R10,L+R20,L+R50 were incubated with LPS for 20 min and 12 hours.MTT were used to detect cell proliferation .The protein level of ERK1/2, and phosphorylation of ERK 1/2 was measured by Western blot respectively .Results Compared with control , LPS signifi-cantly reduced cell proliferation (P<0.05), increased(P<0.01)cell death, and up-regulated the level of phosphorylation of ERK 1/2(P<0.01).Pretreatment of resveratrol attenuated the inhibition of LPS on cell viability and down -regulated the level of phosphoryla-tion of ERK1/2.Conclusion Resveratrol may exert its cytoprotection effects on LPS -induced H9c2 cells via down regulating the activa-tion of p-ERK1/2

目的 建立2型糖尿病患者胱抑素C肾小球滤过率(GFR)估算公式,并对其预测肾功能状态的适用性进行评价.方法 收集856例2型糖尿病患者相关资料,检测血清肌酐(SCr)、半胱氨酸蛋白酶抑制剂C(Cys C)及糖化血红蛋白(HbA1c)浓度,同时行同位素99m锝-二乙三胺五乙酸(99mTc-DTPA)肾动态显像,得出同位素GFR(iGFR),并以iGFR作为GFR参考标准.从856例患者中随机选取660例用于建立糖尿病人群估算公式,其余196例作为验证组,采用配对t检验、Bland-Altman一致性检验、x2检验、Spearman相关分析、ROC曲线等统计学方法对自建公式与简化肾脏病膳食改良试验(MDRD)公式、慢性肾脏病流行病学协助组Cys C公式和SCr/Cys C联合公式(CKD-EPIcysc、CKD-EPIscr/Cysc)估算的GFR (eGFR)进行比较.结果 糖尿病患者基于Cys C建立的公式为87.2×Cys C-1-4.6.在验证组中,自建公式估算eGFR与iGFR比较,其偏差、一致性及符合率均显著优于MDRD和CKD-EPISCr/CysC公式,偏差和符合率优于CKD-EPICysC公式.自建公式和CKD-EPICysC公式对肾功能轻中度下降的糖尿病患者进行慢性肾脏病正确分期的百分比显著高于MDRD和CKD-EPISCr/CysC公式.自建公式和CKD-EPICysC公式在不同血糖控制水平患者间的偏差和符合率比较,差异无统计学意义(P＞0.05).结论 自建Cys C公式在估算不同肾功能水平和不同血糖控制水平的糖尿病患者GFR时均表现出较好适用性.

Kruppel样因子15(KLF15)是Kruppel样转录因子家族中的一员.Kruppel样转录因子家族特征性结构是含有3个Kruppel样锌指结构,与DNA的CACCC元件和富含GC区连接,从而调控转录激活或抑制.KLF15在许多生物过程中起重要作用,包括细胞的增殖、分化、发展和凋亡.研究证实,KLF15参与调节三大物质代谢:糖代谢、脂肪酸代谢、氨基酸代谢.在循环系统方面,KLF15过表达能够抑制心肌肥厚,而KLF15的缺乏会引起心力衰竭、主动脉瘤的产生.此外,KLF15在肾病、肾纤维化、骨骼肌脂质利用、昼夜节律等诸多方面起重要作用.随着对KLF15功能的认识越来越深入,KLF15有望成为有效治疗致死性疾病的新靶点.

目的:研究白藜芦醇是否通过调节p38MAPK活性来抑制脂多糖诱导的H9c2细胞死亡.方法:将H9c2细胞分为对照组(Con)组,脂多糖(LPS)组,白藜芦醇(Res)组,脂多糖+白藜芦醇(RL)组.其中Res组和RL组分别予5 μmol/L白藜芦醇预先处理,然后给予LPS组和RL组10 μg/mL脂多糖共同孵育30 min和12 h,MTT法检测细胞增殖,流式细胞术检测细胞死亡,Western blot检测p38MAPK磷酸化水平.结果:LPS使细胞增殖明显减少(P ＜ 0.05),死亡数明显增加(P ＜ 0.01),p38MAPK磷酸化水平显著升高(P ＜ 0.01).预先给予白藜芦醇处理后,不仅细胞增殖明显增加(P ＜ 0.05),细胞死亡数显著降低(P ＜ 0.01),而且p38MAPK磷酸化水平明显下降(P ＜ 0.01).结论:白藜芦醇可能通过下调p38MAPK磷酸化抑制脂多糖诱导的H9C2细胞死亡,从而发挥其保护细胞的作用.

Objective To compare blood glucose levels between patients with type 1A diabetes treated with autologous hematopoietic stem cell transplantation (AHSCT) or insulin therapy by continuous glucose monitoring system (CGMS). Methods Patients with newly-diagnosed type 1A diabetes were divided into AHSCT group (n=22) and insulin group (n=22). C-peptide area under curve (AUCC), glycosylated hemoglobin (HbA1c) and daily dose of insulin were recorded. CGMS was used to measure mean blood glucose (MBG), standard deviation of blood glucose (SDBG), largest amplitude of glycemic excursions (LAGE), mean amplitude of glycemic excursions (MAGE), mean of daily difference (MODD), low blood glucose index (LBGI) and the percentage of time at hyperglycaemia at each follow-up visit. Results There were no significant differences in HbA1c, AUCC or blood glucose fluctuation coefficient (BGFC) levels between two groups before treatment (all P>0.05). The level of HbA1c was significantly decreased 24 months after treatment in all patients, but HbA1c, LAGE and LBGI were not significantly different between groups (all P>0.05); AUCC, MBG, SDBG, MODD and MAGE in the AHSCT group were significantly superior to that in the insulin group (P<0.01, and 0.05). Daily dose of insulin in the AHSCT group was significantly less than that in the insulin group (P<0.01). The percentage of time at hyperglycaemia (blood glucose >11.1 mmol/L) in the AHSCT group was significanlty lower than that in the insulin group (P<0.05). The percentage of time of blood glucose between 3.9 and 11.1 mmol/L in the AHSCT group was significanlty higher than that in the insulin group (P<0.05). There was no statistical difference in hypoglycemia (blood glucose<3.9 mmol/L) between the two groups (P>0.05). Conclusion AHSCT can significantly improve glucose control and reduce dosage of insulin and overall glucose excursions as compared with insulin therapy.

Objective To investigate the applicability of using Cys C-based formulas for estimation of GFR in Chinese type 2 diabetic patients.Methods Firstly,the accuracy of two assays(PETIA and PENIA) for Cystatin C was evaluated.Then 158 type 2 diabetic patients were collected for determination of serum Cystatin C level and renal clearance of 99mTc-DTPA,which was used as the reference of glomerular filtration rate.Cystatin C was measured by PETIA and PENIA,and the results of eGFR calculated by Grubb formula,Stevens formula,Rule formula and Macisaac formula were compared with the isotopic GFR(iGFR) by paired t test,and bias,15%,30%,50% coincidences were analyzed.Results PETIA and PENIA had good correlation(r=0.986),the bias at medical decision level was acceptable.The 99mTc-DTPA clearance was(96.42±28.93) mL·min-1·(1.73m2)-1,and the median level of Cystatin C was 0.85 mg/L(0.46-4.32 mg/L).eGFR calculated by all the Cys C-based formulas had correlation with iGFR.The eGFR calculated by Rule formula and Macisaac formula had smaller bias with iGFR than that calculated by Grubb formula and Stevens formula,and the 15%,30% and 50% coincidences were different between the 4 formulas.When iGFR≥90 mL·min-1·(1.73m2)-1,the 15% coincidence for Macisaac formula reached 65%.Conclusions The four Cys C-based formulas have different results when used for calculating eGFR in type 2 diabetic patients.All the factors,including ethnicity,age and kind of disease should be considered for selecting the appropriate formula to calculate the eGFR.

Objective To investigate the suitable equation of glomerular filtration rate(GFR) in estimating renal function for type 2 diabetes mellitus patients,and to compare the application effects of modification of diet in renal disease(MDRD),reexpressed modification of diet in renal disease(rMDRD),chronic kidney disease epidemiology collaboration(CKD-EPI) and cystatin C(Cys C)-based equations for GFR.Methods Clinical data from 178 type 2 diabetes mellitus patients were collected for the detection of serum creatinine(SCr) level,Cys C level and renal clearance of 99mTc-diethylenetriamine pentaacetic acid-deoxyglucose(99mTc-DTPA) which was used as the reference of isotopic GFR(iGFR).According to the standard of the American Diabetes Association(ADA),the 178 patients were classified into 3 groups.The equations for estimated GFR(eGFR) were compared with iGFR by paired t test,linear analysis,Bland and Altman procedures,receiver operating characteristic(ROC) curves and 15%,30% and 50% coincidences.Results When GFR<60 mL/(min·1.73 m2),there was no significant difference between eGFR and iGFR in MDRD equation,but when GFR≥60mL/(min·1.73 m2),there was significant difference(P<0.001).There was no significant difference of eGFR for rMDRD equation in accuracy and diagnostic sensitivity.When GFR≥90 mL/(min·1.73 m2),the coincidences were higher in CKD-EPI equation than those in MDRD and rMDRD equation,but when GFR 60-89 mL/(min·1.73 m2),they were lower than those in Cys C GFR equation.The Cys C GFR equation had a better correlation with eGFR and iGFR,a less bias,a higher deviation and a higher coincidence than those in MDRD and rMDRD equations,especially when GFR≥90 mL/(min·1.73 m2),only there was no significant difference between eGFR and iGFR,and Cys C GPR equation was better than the other 3 equations.Conclusions The 4 equations can estimate GFR accurately for type 2 diabetes mellitus patients when GFR<60 mL/(min·1.73 m2),but when GFR≥90 mL/(min·1.73 m2),Cys C GFR equation shows a significant superiority than the old SCr based equations.