结缔组织病患者骨代谢的变化及云克对类风湿关节炎骨代谢的影响

目的：通过测定骨密度和骨代谢指标了解结缔组织病患者骨质疏松的发生情况和骨代谢的变化情况,并评价云克治疗类风湿关节炎（rheumatoid arthritis,RA）对骨代谢的影响。方法：选取2016年2月—2017年2月于江苏大学附属医院风湿免疫科住院患者共280例,其中系统性红斑狼疮（systemic lupus erythematosus,SLE）75例,原发性干燥综合征（primary Sjogren syndrome,pSS）45例,RA 160例。采用双能X线法测定患者腰椎和髋部骨密度,采用电化学发光免疫分析法检测骨钙素（osteocalcin,OC）、Ⅰ型原胶原N端前肽（N?terminal propeptide of typeⅠ procollagen,PINP）和Ⅰ型胶原C端肽交联（C?terminal cross?linking telopeptide of type Ⅰ collagen,CTX）。分别分析骨代谢指标OC、PINP和CTX与患者年龄、病程、体重指数（body mass index,BMI）、C?反应蛋白（C?reactive protein,CRP）、红细胞沉降率（erythrocyte sedimentation rate,ESR）、补体C3、类风湿因子（rheumatoid factor,RF）、骨密度以及激素使用程度等因素的相关性。有70例RA患者给予云克治疗4个月,统计分析云克治疗前后骨代谢指标的差异。结果：280例结缔组织病患者骨量减少的发生率为27.7%,骨质疏松的发生率为21.5%。与SLE组比较,OC、PINP以及CTX水平在pSS组和RA组呈升高趋势;同时,pSS和RA患者的OC、PINP以及CTX水平在骨量正常组、骨量减少组和骨质疏松组呈逐渐升高的趋势。相关性分析发现,pSS患者骨代谢指标与RF呈显著正相关（OC：r=0.570,P=0.002;PINP：r=0.752,P < 0.001;CTX：r=0.660,P < 0.001）;SLE患者OC、PINP与髋部骨密度呈显著负相关（OC：r=-0.382,P=0.028;PINP：r=-0.527,P=0.002）;pSS患者OC、CTX与髋部骨密度呈显著负相关（OC：r=-0.471,P=0.013;CTX：r=-0.422,P=0.028）。与治疗前比较,云克治疗4个月后RA患者OC水平上升（P < 0.05）,ESR水平下降（P < 0.05）;而PINP、CTX、CRP和骨密度无明显变化。结论：结缔组织病患者存在不同程度的骨代谢异常,骨质疏松发生率较高。骨代谢指标可早期预测骨量丢失情况。云克治疗有助于改善RA患者骨代谢。     

课堂教学语言艺术

本文针对中等医学教育的特点，对课堂教学语言艺术的概念、意义、要求和特征以及它对教学信息传递所起的作用、效果作了论述，说明教师要创造教学语言艺术，用之于优化教学目标。     

全反式维甲酸对肝癌细胞连接蛋白基因表达及细胞间隙连接通讯功能的影响

目的 观察肝癌细胞株SMMC-7721和BEL-7404经全反式维甲酸(ATRA)诱导前后CX26、CX32、CX43基因在转录水平的改变以及对细胞间隙连接功能的影响。方法 分别用常规培养基和含ATRA浓度为10～mol／L的培养基培养SMMC-7721和BEL-7404两种肝癌细胞株细胞,于药物诱导细胞24、48、72h时,收集各组细胞并提取总mRNA,RT-PCR法检测CX26、CX32、CX43基因表达的情况。通过染料传输实验,观察细胞间隙连接通讯功能的变化。结果 SMMC-7721细胞和BEL-7404细胞在ATRA处理前没有CX26 mRNA和CX32 mRNA的表达,但均有CX43 mRNA的表达。经ATRA处理后出现CX26 mRNA和CX32 mRNA的表达。经ATRA处理后SMMC-7721细胞间隙连接通讯功能增强,而且随ATRA作用时间延长,细胞间隙连接通讯功能增强越明显。BEL-7404细胞经ATRA处理24、48、72h后细胞间隙连接通讯功能没有明显的改变。结论 全反式维甲酸能够在转录水平上调CX26、CX32基因在肝癌细胞SMMC-7721和BEL-7404中的表达。肝癌细胞SMMC-7721在CX26、CX32基因转录水平上调的同时伴有细胞间隙连接通讯功能的增强。肝癌细胞BEL-7404在CX26、CX32基因转录水平上调的同时不伴有细胞间隙连接通讯功能的增强。这说明两种细胞的间隙连接通讯功能调节机制不同。     

大鼠脾脏形态学的年龄变化

本文用组织学和组织化学方法对SD大鼠脾脏的年龄变化进行了观察。大鼠脾脏中白髓随增龄而逐渐减少,结构紊乱,淋巴细胞密度下降。白髓和红髓分界不清。巨噬细胞及中性粒细胞数量增多。ANAE、ACP、AlP活性随增龄而加强。ATP酶无明显变化,SDH、LDH和5-Nase活性下降,说明老龄鼠脾脏细胞中的代谢活动发生了一定的变化。     

献血员HBV携带状况的筛检方法研究

将应用常规RPHA法检查HBsAg阴性的1074份献血员血清再用ELISA和SPRIA法检测。其中又查出HBsAg、HBeAg和抗-HBc-IgM标志,其阳性率分别达5.96％,2.14％和3.35％,HBV携带率竟高达9.68％。说明用常规RPHA法筛选出的HBsAg阴性献血员仍有传播乙型肝炎的危险。用ELISA法并联检测HBsAg和抗-HBc-IgM,在RPHA法检查HBsAg阴性的献血员中、其阳性率为8.29％,敏感度达85.58％,因而这种并联筛检方法可成为一种切实可行的筛检方法。     

乳腺癌发生模型MCF10各阶段细胞系中TIMP3基因启动子区甲基化分析

目的探讨抑癌基因TIMP3失活与乳腺癌发生和进展的关系。方法用甲基化特异性PCR技术和亚硫酸盐测序技术检测乳腺癌发生模型MCF10的增生细胞系MCF10A、癌前细胞系MCF10AT、导管内癌细胞系MCF10DCIS.com、浸润癌细胞系MCF10CA1a和转移癌细胞系MCF10CA1d、MCF10CA1h中TIMP3启动子区甲基化状态。结果甲基化特异性PCR分析显示,在上述各细胞系中,TIMP3启动子区均呈高度甲基化状态。亚硫酸盐测序显示,在上述各细胞系中,测序区内的68个CG位点几乎全部发生了甲基化,且甲基化累及了绝大部分等位基因。结论TIMP3基因启动子区甲基化在乳腺癌的发生和进展中起重要作用,可能成为早期诊断乳腺癌和判断乳腺癌预后的分子生物学标记。     

p38丝裂素活化蛋白激酶信号转录对IL-1β诱导肾小管细胞转分化的影响

目的　探讨 p38MAPK信号转导途径对IL 1β诱导肾小管上皮细胞转分化的影响及其导致的功能改变。方法　以培养的人近端肾小管上皮细胞 (HK 2 )为研究对象 ,给予IL 1β(10ng/ml)刺激 2 4h ,在阻断实验中加入SB2 0 35 80阻断 p38通路。采用蛋白印记杂交法检测 p38MAPK的磷酸化程度 ;细胞表型特征检测包括细胞形态变化、标志蛋白 (角蛋白、α SMA)的表达及分布、细胞增殖、黏附和移行能力。结果　 (1)IL 1β刺激 6～ 4 8h可使α SMA表达上调 ,刺激 2 4h可使角蛋白表达减弱、α SMA分布紊乱 ,但细胞形态变化不明显 ;(2 )IL 1β在 5min时可使p38激活达高峰 ,较基础水平升高 2～ 3倍 (P <0 0 5 ) ;至 12 0min时 p38则呈持续激活状态 ;(3)p38阻断剂 (SB2 0 35 80 )对HK 2细胞α SMA的基础表达有抑制作用 ,抑制率 37 13% (P <0 0 1) ;同时可明显抑制IL 1β刺激的α SMA表达 ,抑制率 4 7 0 7% (P <0 0 0 1) ;(4)IL 1β及 p38阻断剂并不影响HK 2细胞之间的黏附能力 ,但IL 1β可使细胞的移行能力增强 (P <0 0 1) ,而 p38阻断剂可明显抑制其移行能力。IL 1β并不影响HK 2细胞增殖。结论　IL 1β可以激活肾小管上皮细胞p38/MAPK ,并可使其发生表型转化 ,这一作用部分通过 p38信号通路所介导。     

体外辐射模拟P53依赖的小鼠肠干细胞损伤研究

目的 建立6 Gy小鼠肠干细胞体外辐射模型,模拟在体P53依赖的肠干细胞辐射损伤修复过程.方法 分离C57BL/6(野生型)、P53-/-小鼠小肠隐窝,分别于基质胶内接种进行3D培养,24 h后予X线6 Gy照射.比较两组隐窝克隆出芽过程和照后6d克隆存活率.对野生型隐窝克隆用RT-PCR检测照后24 h内不同时间点P53靶基因Bax、Bbc3、Mdm2、P21、Bcl2l1以及肠干细胞特征性基因Lgr5、Olfm4、Ascl2的表达.结果 C57BL/6和P53-/-小鼠小肠隐窝于接种后48 h出芽,在接种后96 ～ 120 h形成类器官样结构.6 Gy辐照可诱导C57 BL/6隐窝细胞24 h内大量死亡,完全抑制克隆出芽,仅(43.84 ±5.73)％的克隆存活;而对P53-/-小鼠隐窝出芽时间无影响,且有(83.69±5.89)％隐窝克隆存活,显著高于对照组(P＜O.01).在野生型隐窝克隆中,P53下游靶基因的表达可被辐射迅速激活,并在24 h内维持高水平,而干细胞特征性基因在12h迅速下降,24 h到达最低水平.结论 6 Gy照射小鼠小肠隐窝能较好地模拟在体P53依赖的促凋亡因素对肠干细胞的损伤.     

Trends in etiologies of chronic pancreatitis within 20 years: analysis of 636 cases

Background  The prevalence of chronic pancreatitis has increased during recent years in Asia-Pacific areas as well as in China. The etiologies vary in different regions and periods. This study aimed to investigate the changing etiologies of chronic pancreatitis within 20 years at Peking Union Medical College Hospital in China.

Methods  Retrospective analysis of the etiologies of 636 cases of chronic pancreatitis at Peking Union Medical College Hospital from 1990 to 2010 was performed. Patients were divided into two groups according to two time periods (1990–2000 and 2001–2010). Statistical analysis was performed using the chi-square test.

Results  The morbidity rate of chronic pancreatitis in China has recently increased. The main etiology changed from biliary diseases in the 1990s (decreased from 36.8% to 28.1%) to alcohol abuse after the year 2000 (increased from 26.5% to 36.8%). The main etiology of biliary diseases is stones in the cholecyst or bile duct, and the percentage of cholecystitis cases has increased. Autoimmune disease, including autoimmune pancreatitis, has increased quickly and currently accounts for 7.3% of cases because a greater number of autoimmune pancreatitis cases are being diagnosed. Approximately 9.5% of chronic pancreatitis cases are caused by multiple factors such as alcohol abuse and bile duct stones. Other factors include cholecystectomy and acute pancreatitis.

Conclusions  The main etiology of chronic pancreatitis has changed from biliary disease to alcohol abuse in recent years. Autoimmune factors have also obviously increased.

     

Chemokine-like factor 1, a novel cytokine, contributes to airway damage, remodeling and pulmonary fibrosis

Background Chemokine-like factor 1 (CKLF1) was recently identified as a novel cytokine. The fulllength CKLF1 cDNA contains 530 bp encoding 99 amino acid residues with a CC motif similar to that of other CC family chemokines. Recombinant CKLF1 exhibits chemotactic activity on leucocytes and stimulates proliferation of murine skeletal muscle cells. We questioned whether CKLF1 could be involved in the pathogenesis of inflammation and proliferation in the lung. Therefore we used efficient in vivo gene delivery method to investigate the biological effect of CKLF1 in the murine lung. Methods CKLFl-expressing plasmid, pCDI-CKLF1, was constructed and injected into the skeletal muscles followed by electroporation. Lung tissues were obtained at the end of week 1,2,3 and 4 respectively after injection. The pathological changes in the lungs were observed by light microscope. Results A single intramuscular injection of CKLF1 plasmid DNA into BALB/c mice caused dramatic pathological changes in the lungs of treated mice. These changes included peribronchial leukocyte infiltration, epithelial shedding, collagen deposition, proliferation of bronchial smooth muscle cells and fibrosis of the lung. Conclusions The sustained morphological abnormalities of the bronchial and bronchiolar wall, the acute pneumonitis and interstitial pulmonary fibrosis induced by CKLF1 were similar to phenomena observed in chronic persistent asthma, acute respiratory distress syndrome and severe acute respiratory syndrome. These data suggest that CKLF1 may play an important role in the pathogenesis of these important diseases and the study also implies that gene electro-transfer in vivo could serve as a valuable approach for evaluating the function of a novel gene in animals.