目的:探讨当归通痹汤联合中药热敷治疗膝关节骨性关节炎的疗效及对白细胞介素(IL)-1β、基质金属酶(MMP)-3和肿瘤坏死因子(TNF)-α水平的影响.方法:选取我院收治的膝关节骨性关节炎患者140例,随机分为两组各70例.对照组给予中药热敷治疗,治疗组在对照组基础上给予当归通痹汤,对比两组治疗后的临床疗效、VAS评分、WOMAC指数和ADL评分,以及IL-1β、MMP-3和TNF-α水平.结果:治疗后,治疗组的总有效率为94.29％,高于对照组的81.43％,差异有统计学意义(P<0.05);两组ADL评分明显升高,WOMAC指数、VAS评分明显降低(P<0.05);且治疗组比对照组明显改善(P<0.05);两组患者治疗后的IL-1β、MMP-3和TNF-α水平均显著低于治疗前(P<0.05);且治疗组低于对照组,差异有统计学意义(P<0.05).两组患者不良反应发生率差异无统计学意义(P>0.05).结论:膝关节骨性关节炎患者采用当归通痹汤联合中药热敷治疗,具有较好的临床疗效,能够改善患者膝关节功能,值得在临床上推广应用.

目的 对文献报道的脑瘫差异表达基因进行生物信息学分析,并筛选脑瘫核心驱动基因.方法 通过检索Pubmed、中国知网数据库相关文献,获取脑瘫差异表达基因,并进行GO功能富集分析、KEGG信号通路分析和通路串话分析,应用STRING和Cytoscape软件构建蛋白质互作网络(PPI),并筛选排名前10的差异表达基因,即为脑瘫核心驱动基因.结果 得到298个脑瘫差异表达基因.GO功能富集分析结果显示,脑瘫差异表达基因在生物学过程方面主要与免疫应答、防御反应、对含氧化合物的反应等有关,在细胞组分方面主要与轴突、突触后膜、突触等有关,在分子功能方面主要与蛋白聚糖结合、胶原结合、抗原结合等有关.KEGG信号通路分析结果显示,脑瘫差异表达基因主要参与Th17细胞分化、Toll样受体信号通路、NOD样受体信号通路、PI3K-Akt信号通路等.脑瘫差异表达基因的通路串话关系网络中,T细胞受体信号通路、IL-17信号通路、细胞因子和细胞因子受体相互作用等通路与其他的通路之间存在较多的串话关系.PPI网络中的10个关键基因是IL-6、TNF、IFNG、CXCL8、TLR4、ICAM1、IL-4、IL-1β、CCL5、PTGS2.结论 脑瘫差异表达基因可能通过免疫应答等生物学过程发挥作用,Th17细胞分化、T细胞受体信号通路等信号通路参与了脑瘫的发生发展;IL-6、TNF、IFNG等可能是脑瘫发生发展的核心驱动基因.

肺癌是最常见恶性肿瘤之一,其发病率、死亡率高,且呈上升趋势.咳嗽是其最常见症状之一,也多为首发症状,且肺癌的咳嗽各具特点,临证时黄金昶教授通过抓主证的方法进行辩治,取得较好的临床效果.本文疏理肺癌患者常见的咳嗽类型,总结黄金昶教授治疗肺癌患者咳嗽的临床经验.黄金昶教授指出了肺癌患者临床最常见的九种咳嗽,包括因痒作咳,卧则咳嗽,动则咳剧,饮热呛咳,因痰而咳,剧烈阵咳,咳痰无力,脏结实咳,慢性咳嗽等,并分析相关病因病机,提出相应的治疗方案.黄金昶教授治疗肺癌咳嗽经验独到,疗效显著,值得学习与临床推广.

背景:前期研究发现,骨碎补总黄酮可促进纳米骨材料表面MC3T3-E1细胞的成骨分化,其作用机制有待进一步研究.目的:探究骨碎补总黄酮联合纳米骨材料对MC3T3-E1细胞发挥作用的机制.方法:将MC3T3-E1细胞与纳米骨材料共培养,选取100 mg/L和250 mg/L骨碎补总黄酮进行药物干预,以10μg/L转化生长因子β刺激为阳性对照组.分组如下:①正常组;②DKK1组:Wnt通路抑制剂DKK1(0.1 mg/L)阻断Wnt/β-catenin信号通路;③DKK1+转化生长因子β组;④DKK1+100 mg/L骨碎补总黄酮组;⑤DKK1+250 mg/L骨碎补总黄酮组;⑥DKK1+纳米骨+转化生长因子β组;⑦DKK1+纳米骨+100 mg/L骨碎补总黄酮组;⑧DKK1+纳米骨+250 mg/L骨碎补总黄酮组.在干预24,48 h后收获细胞,免疫荧光双染法观察Wnt/β-catenin通路中Wnt与LRP结合情况,Real-time PCR和Western blot检测β-catenin、LRP5、Gsk-3β、Cyclin D1、RUNX2的表达.结果与结论:①激光共聚焦扫描显微镜下显示DKK1+转化生长因子β组、DKK1+250 mg/L骨碎补总黄酮组、DKK1+纳米骨+转化生长因子β组、DKK1+纳米骨+250 mg/L骨碎补总黄酮组棕黄色染色较明显,表明Wnt与LRP结合较其他组更好;②Real-time PCR和Western blot结果显示,骨碎补总黄酮可促进β-catenin、LRP5、RUNX2的表达,下调GSK-3β的表达,说明骨碎补总黄酮通过激活经典Wnt/β-catenin信号通路促进成骨细胞增殖分化,且骨碎补总黄酮诱导的基因活化呈剂量依赖性.

目的 探讨复发性阿弗他溃疡(recurrent ahthous ulcer,RAU)患者唾液钙卫蛋白水平与疾病活动期的关系.方法 RAU患者117例,其中活动期61例为活动期组,间歇期56例为间歇期组;同期58例体检健康者为对照组.检测3组白细胞计数、空腹血糖、三酰甘油、总胆固醇、血肌酐及高敏C反应蛋白水平,测定3组唾液钙卫蛋白水平,多因素logistic回归分析各指标对RAU活动期的影响.结果 活动期组唾液钙卫蛋白[(25.85±4.50)mg/L]、白细胞计数[(7.51±2.80)×109/L]、高敏C反应蛋白[(1.95±1.26)mg/L]水平高于间歇期组[(21.00±3.33)mg/L、(6.49±1.49)×109/L、(1.37±1.23)mg/L]和对照组[(20.57±2.95)mg/L、(6.37±1.80)×109/L、(1.41±1.03) mg/L] (P＜0.05),间歇期组与对照组比较差异无统计学意义(P＞0.05);3组空腹血糖、三酰甘油、总胆固醇、血肌酐水平比较差异无统计学意义(P＞0.05);多因素logistic回归分析结果显示,白细胞计数增高(OR=1.339,95％CI:1.050～1.709,P=0.019)、唾液钙卫蛋白增高(OR=1.382,95％CI:1.209～1.581,P＜0.001)是RAU活动期的影响因素.结论 高唾液钙卫蛋白、高白细胞计数与RAU活动期相关,可用于评估RAU病情和临床疗效.

目的:探讨血清胃泌素释放肽前体(pro-gastrin-releasing peptide,ProGRP)、甲胎蛋白(α-fetoprotein,AFP)、糖类抗原199(carbohydrate antigen 199,CA199)、糖类抗原125(carbohydrate antigen 125,CA125)、人附睾分泌蛋白4(human epididymal protein 4,HE4)单项检测及ProGRP、AFP、CA199联合平行检测对卵巢癌的诊断价值.方法:回顾性分析379例疑诊卵巢癌者的临床病例资料,其中123例患者经病理学明确诊断为卵巢癌(卵巢癌组),另256例为非卵巢癌患者(非卵巢癌组).比较2组患者的血清ProGRP、AFP、CA199、CA125、HE4水平,探讨血清ProGRP、AFP、CA199、CA125、HE4单项检测及ProGRP、AFP、CA199联合检测诊断卵巢癌的效能,绘制基于血清ProGRP、AFP、CA199、CA125、HE4单项检测及ProGRP、AFP、CA199联合检测诊断卵巢癌的受试者工作特征曲线(receiver operating characteristic curve,ROC曲线),计算ROC曲线下面积(area under the ROC curve,AUC).结果:卵巢癌组Ⅰ期患者的血清ProGRP、AFP、CA199水平均明显低于Ⅱ～Ⅳ期患者(P＜0.05);卵巢癌组的血清ProGRP、AFP、CA199、CA125及HE4水平均明显高于非卵巢癌组(P＜0.05);血清ProGRP、AFP、CA199、CA125、HE4单项检测诊断卵巢癌的最佳临界值分别为69.21 pg/mL、28.95 ng/mL、42.38 U/mL、57.65 U/mL、125.64 pmol/L,血清ProGRP、AFP、CA 199、CA125、HE4单项检测及ProGRP、AFP、CA199联合平行检测诊断卵巢癌的AUC分别为0.786、0.821、0.897、0.912、0.926及0.948,灵敏度分别为78.05％、80.49％、85.37％、88.62％、91.06％及77.24％,特异度分别为85.94％、86.72％、87.89％、90.63％、90.23％及97.27％,准确率分别为83.37％、84.70％、87.07％、89.97％、90.50％及90.77％.结论:血清ProGRP、AFP、CA199、CA125、HE4单项检测及ProGRP、AFP、CA199联合检测在卵巢癌诊断中均具有良好的应用价值,但ProGRP、AFP、CA199联合平行检测的可靠性更佳,可用于卵巢癌筛查工作中.

背景:前期研究发现,骨碎补总黄酮可促进纳米骨材料表面MC3T3-E1细胞的成骨分化,其作用机制有待进一步研究.目的:探讨骨碎补总黄酮联合纳米骨材料对MC3T3-E1细胞成骨分化的影响及作用机制.方法:将MC3T3-E1细胞接种于纳米骨材料表面,分别加入含0,100,250 mg/L骨碎补总黄酮的培养基培养.培养第1,3,5,7,9,11,14天,检测细胞碱性磷酸酶活性;培养第1,2,3,4周,ELISA法检测培养上清中骨钙蛋白水平;培养第7,14,21天,茜素红S染色观察细胞钙化结节形成情况;培养第14天,PCR检测成骨相关基因表达.结果 与结论:①第1,3,14天,3组细胞碱性磷酸酶活性比较差异无显著性意义(P＞0.05);培养第5,7,9天,100,250 mg/L组碱性磷酸酶活性均高于0 mg/L组(P＜0.05),100 mg/L组、250 mg/L组间比较差异无显著性意义(P＞0.05);②培养第1,2,3,4周,100,250 mg/L组骨钙蛋白质量浓度均高于0 mg/L组(P＜0.05),100 mg/L组、250 mg/L组间比较差异无显著性意义(P＞0.05);③培养第7,14,21天,100,250 mg/L组可见明显的钙化结节;④培养第14天,100,250 mg/L组骨钙蛋白、Ⅰ型胶原和骨桥蛋白mRNA表达水平均高于0 mg/L组(P＜0.05),100 mg/L组、250 mg/L组间骨钙蛋白、Ⅰ型胶原和骨桥蛋白mRNA表达比较差异无显著性意义(P＞0.05);⑤结果表明,骨碎补总黄酮联合纳米骨材料可诱导MC3T3-E1细胞分化为成骨细胞,其机制可能是通过提高碱性磷酸酶活性、骨钙蛋白水平及Ⅰ型胶原、骨钙蛋白和骨桥蛋白mRNA表达来实现的.

目的 探讨唾液中白细胞介素-8(interleukin-8,IL-8)的表达情况及其与复发性阿弗他溃疡(recurrent ahthous ulcer,RAU)的相关性.方法 该研究共纳入117名RAU患者及58名正常对照者,RAU患者又根据疾病的活动性情况分为活动期组(n=61)及间歇期组(n=56).采用酶联免疫吸附(ELISA)法测定并比较各组的唾液IL-8水平,采用多元Logistic回归分析评价各组唾液IL-8水平与RAU的相关性.结果 活动期RAU患者唾液IL-8水平较间歇期组显著升高[(46.28±19.64)pg/ml vs(27.02±12.27)pg/ml,P<0.001];间歇期组RAU患者唾液IL-8水平较对照组显著升高[(27.02±12.27)pg/ml vs(6.02±1.95)pg/ml,P<0.001].多元Logistic回归分析显示唾液IL-8水平是RAU活动期的独立危险因子(优势比1.071,95％CI 1.042-1.100,P<0.001).结论 RAU患者活动期唾液IL-8水平显著升高,唾液IL-8水平是预测RAU活动性的独立危险因子.

The inflammatory cytokines of IL-1β and TNF-α participate in the process of intervertebral disc degeneration is focused by the spine surgeons. Inflammatory cytokines represented by IL-1β and TNF-α act as the key factors in the process of intervertebral disc degeneration resulting in low back pain and radicular symptoms. Annulus fibrosus and nucleus pulposus can secrete TNF-α and IL-1β under the stimulation of mechanical injury, overstressed, genetic susceptibility and infection. The mechanism of inflammatory cytokines in intervertebral disc degeneration needs further investigation. The emphasis of researches will be on the inflammatory cytokines in the regulation of mechanism in the intervertebral disc degeneration, molecular targeted therapy, cell signaling pathways and the best time of anti-inflammatory therapy providing more evidence in the clinical application.

[Objective]The purpose of the study was to establish the utility of a compression chamber and examine the effects of various magnitudes and durations of compression on rabbit vertebral cartilage endplate cells,particularly,their influence on the expression of inflammatory,catabolic,and anabolic genes.[Method]A chamber capable of imparting 0 to 5 MPa of hydrostatic compression onto the cartilage endplate cells was fabricated. Healthy rabbit cartilage endplate cells were cultured. After morphological and functional identification,the cells were exposed to static compression at 0. 01 MPa,0. 7 MPa,2 MPa,and 4 MPa for 4 or 24 h. Gene expression analysis( real- time polymerase chain reaction) was performed to compare markers of inflammation( inducible nitric oxide synthase,cyclooxygenase- 2),matrix catabolism( matrix metalloproteinase- 3),and anticatabolic / anabolic metabolism( tissue inhibitor of metalloproteinase- 1,aggrecan) in control and compressed cells. [Result] Compression resulted in magnitude- and duration- dependent changes in gene expression. Decreasing magnitudes and times effected more changes in the expression of anticatabolic / anabolic metabolism genes,whereas increasing duration resulted in increases in catabolic gene expression. [Conclusion]These data demonstrate the favorable effects of compression on genes involved in matrix homeostasis and catabolic activity in response to sustained loading levels. In addition,the data provide an improved understanding of how compression affects cell signaling,which has the potential to be exploited to initiate repair and prevent disc degeneration.

[Objective]To investigate the effects of apigenin on proteoglycan core protein gene expression in the human lumbar intervertebral disc nucleus pulposus in response to hydrostatic pressure. [Methods]Thirty-two lumbar intervertebral disc nucleus pulposus specimens were obtained from patients who had undergone posterior discectomy.The syringes were placed in a water filled pressure vessel kept at 37℃.Hydrostatic pressures of 1(control),3 and 30 atmospheres(atm) were applied.After incubation of 2 hours,each nucleus pulposus specimen was embedded in paraffin,sliced for situ hybridization to study the effect of the apigenin of the intervertebral disc nucleus pulposus proteoglycan core protein gene's transcription.As a competitive inhibitor of NO synthases,NG-Monomethy-L-arginine(L-NMMA,500 μmol) and as an organic donor of NO,S-nitroso-N-acetylpenicillamine(SNAP,200 μmol) were used.[Results]The results of apigenin 3 atm group and the L-NMMA 3 atm group in situ hybridization to proteoglycan core protein showed the largest number of positive nucleus pulposus cells,with a largest average optical density.[Conclusion]For the degenerative intervertebral disc nucleus pulposus,apigenin plays an important role in the promotion of proteoglycan core protein mRNA gene's transcription levels.

背景:骨髓间充质干细胞具有多向分化能力,是目前极具潜力的种子细胞及基因治疗的靶细胞。目的:体外培养兔骨髓来源间充质干细胞,了解其生物学特性。方法:采用密度梯度离心法培养兔骨髓间充质干细胞,对细胞的形态及生长特性进行观察,应用流式细胞仪检测细胞表面抗原CD29、CD34、CD44、CD45、CD166、HLA-DR表达,并进行相关生物学特性鉴定。结果与结论:密度梯度离心法能分离培养出纯度较高的骨髓间充质干细胞,流式细胞仪检测骨髓间充质干细胞表达CD29、CD44、CD166,不表达CD34、CD45、HLA-DR。在适当的条件下能够诱导分化成为成骨细胞、软骨细胞等。

BACKGROUND: Tissue engineering repair for blocks of tissue defects needs a high concentration and a large number of cells for incubation. Bone marrow mesenchymal stem cells (BMSCs) are a main source of seed cells, but there are some problems: limitation of number; cell functional degradation. OBJECTIVE: To in vitro isolate, culture and amplify human placenta mesenchymal stem cells (hPMSCs) and human BMSCs, and observe their biological characteristics. DESIGN, TIME AND SETTING: The cytological, in vitro controlled study was performed at the Laboratory of Dongzhimen Hospital from September 2008 to February 2009. MATERIALS: Placenta was supplied by Department of Gynaecology and Obstetrics, General Hospital of Air Force of Chinese PLA. Bone marrow was obtained from healthy adult donors. METHODS: HPMSCs were isolated from placenta of fetation human by density gradient centrifuge. Following bone marrow heparin anticoagulation, human BMSCs were isolated, purified and passaged by density gradient centrifuge. MAIN OUTCOME MEASURES: Cell morphology and growth were observed under an inverted phase contrast microscope. The 5th passage cells were detected by flow cytometry. Differentiation of cells was determined using Von Kossa staining and Oil red O staining. RESULTS: HPMSCs and human BMSCs were both uniformly spindle-shaped in appearance under a microscope. The proliferative ability of hPMSCs were quite strong and declined with passages. After cultured 10 passages in vitro, cell growth became slow, but amplified multiple of hPMSCs was over 2-3 order of magnitude compared with human BMSCs. Both hPMSCs and human BMSCs expressed CD29, CD44 and CD166, but did not express CD34, CD45 and HLA-DR. Both cells had the potential of osteogenic and adipocytic differentiation, but hPMSCs had stronger potential. CONCLUSION: HPMSCs and human BMSCs have similar biological characteristics and the proliferative ability of hPMSCs is stronger than human BMSCs.

Objective To observe the gene expression of collagen BMP-2 (bone morphogenetic protein-2) and TGF-β (transforming growth factor-β) in human thoracic ligamentum flavum.Method BMP-2 and TGF-13 probes were labeled and in situ hybridization results were analyzed by VIDAS system.Result (1) Under 20 years group:the positive gene expression of TGF-β was evident,but it was negative to BMP-2.(2) 21 ~40 years group:there were a few positive signal of both BMP-2 and TGF-β on the insertion part.(3) 41 ~60 years group:there was only few TGF-β positive signal on insertion part,but BMP-2 gene expression was relatively high.(4) Over 61 years group:there was not TGF-β expression,but BMP-2 expression could only be seen on insertion part and junction part,and was low,but the cells were areatus.Conclusion In normal ligamentum,there is expression of TGF-β gene,but it decreases with age;with age advancing BMP-2 emerges and enhances in the insertion part of ligamentum flavum.

目的:椎间盘代谢受许多因素影响,探讨静水压对人体腰椎间盘的一氧化氮产生及蛋白多糖合成的影响。方法:1998-05/2000-03在日本国富山医科药科大学骨科收集后方腰椎椎间盘摘除术所获标本69例样,样品切成1.0～2.0mm碎块后,3与1mL不含碳酸氢钠培养基DMEM一同装入2.5mL塑料注射器中,再放入37℃充满蒸馏水的能够培养椎间盘组织的装置,分别提供0.1,0.3,3MPa气压。蛋白多糖合成由35S-硫酸盐结合率来测定。无活性、稳定的一氧化氮的最终产物亚硝酸盐浓度用分光光度法测定。结果:0.3MPa蛋白多糖合成最高,约为0.1MPa的1.3倍;3MPa抑制蛋白多糖合成率。但静水压对一氧化氮的产生起着逆转效应。在0.3MPa一氧化氮的产生量和0.1MPa的相比略微降低,而3MPa增加一氧化氮的产生量,约是0.1MPa的1.34倍。结论:人的突出腰椎间盘组织诱导一氧化氮产生,其一氧化氮产生和蛋白多糖合成的改变与静水压有明显的相关性。高负荷静水压（3MPa）增加一氧化氮产生量而减少蛋白多糖合成。需要进一步研究澄清在人体腰椎间盘静水压环境下基质金属蛋白酶产生中一氧化氮发挥的作用。如果在椎间盘代谢中一氧化氮的作用得到明了,提示可用特异性抑制诱导型一氧化氮合成酶或一氧化氮的药物阻止或延缓腰椎间盘的退变过程。

目的探讨仿生制备的矿化蚕丝基骨材料在大鼠体内的生物相容性及体内降解性.方法成年雄性SD大鼠16只,体重230～300克.在背部肌肉内和腹腔内植入矿化蚕丝基骨材料,观察时间阶段为2周,4周,6周,8周.结果分别在术后2周,4周,6周,8周,行大体观察,细胞毒性研究,组织学切片观察材料的情况,植入2周后出现炎症反应明显,材料框架基本存在,术后8周大鼠体内植入的矿化蚕丝基骨材料没有明显的排异反应,而且大部分被吸收.结论矿化蚕丝基骨材料具有良好的生物相容性和适宜的生物降解性能,是比较理想的骨缺损的替代材料.