Loss of PPM1A expression enhances invasion and the epithelial-to-mesenchymal transition in bladder cancer by activating the TGF-β/Smad signaling pathway

The transforming growth factor-β (TGF-β) signaling pathway is believed to contribute to carcinoma development by increasing cell invasiveness and metastasis and inducing the epithelial-to-mesenchymal transition (EMT). Protein phosphatase PPM1A has been reported to dephosphorylate TGF-β-activated Smad2/3, thus inhibiting the TGF-β signaling pathway. In this study, we investigated the role of PPM1A in bladder cancer. PPM1A protein expression was analyzed in 145 bladder cancer specimens. The loss of PPM1A expression was predictive of poor survival and high muscle-invasiveness. PPM1A was more commonly deficient among muscle-invasive relapse samples compared to primary tumors in twenty paired bladder cancer tissues. Functional studies indicated that blockade of PPM1A through lentivirus-mediated RNA interference significantly promoted urinary bladder cancer (BCa) cell motility, the EMT in vitro and metastasis in vivo, and these effects were dependent on the TGF-β/Smad signaling pathway. The increase in p-Smad2/3 induced by TGF-β1 correlated with the degree of PPM1A depletion in BCa cells, which resulted in an altered expression profile of TGF-β-inducible genes. The correlations between PPM1A and biomarkers related to the TGF-β signaling pathway and tumor invasion were also detected in BCa samples. These results demonstrate that loss of PPM1A is associated with the development of tumor invasion in bladder cancer.     

颌眉角的最优选择及其在强直性脊柱炎后凸畸形矫形设计中的应用

目的探索不同范围颌眉角对颈椎强直性脊柱炎（ankylosingspondylitis,AS）患者日常生活的影响,确定最优的颌眉角范围,进而为截骨矫形设计提供理论依据。方法选取我科收治的25例颈椎强直的 AS 胸腰段后凸畸形患者,所有患者矫形前后均完成视野相关生活质量功能及期望评分调查问卷表。不考虑手术与否,按照自然站立位颌眉角将所有外观照所对应病例（ n＝50）分为6组：A 组,CBVA＜0°;B组,0°≤CBVA＜10;C 组,10°≤CBVA＜20°;D组,20°≤CBVA＜30°;E 组,30°≤CBVA＜40°;F组, CBVA≥40°。将6组患者每项调查问卷项目进行比较分析,多组间比较时采用Kruskal-Wallis检验,组间两两比较采用Mann-Whitney检验。结果总体评价：C组（10～20）°获得最优的期望值评价,P＜0.05。B、C、D 组（0～30）°获得较其它组更好的功能评价,P＜0.05,但三组间差异无统计学意义。形象评价：A、B、C 组（＜20°）较其它组获得更好的形象满意度及期望值,P＜0.05,但三组间差异无统计学意义。户外活动：A、B、C、D组（＜30°）在多数项目上较其它组获得更好的功能评价及期望值评价。室内活动：C、D组（10～30）°在多数项目上较其它组获得较好的功能评价及期望评价。结论颌眉角在10°～20°时,患者视野相关活动功能及满意度最高,因此,10°～20°为颌眉角的最佳矫正角度。     

肺门作为躯干重心对强直性脊柱炎胸腰段后凸畸形矫形的意义

目的探索能够代表强直性脊柱炎（ankylosingspondylitis,AS）胸腰段后凸畸形躯干重心的影像学标记,进而设计AS胸腰段后凸畸形的截骨矫形方案。方法不规则物体的重心可通过两个不同方向的悬吊或支撑力线获得,通过手术前后髋轴的支撑力线可寻找 AS 胸腰段后凸畸形患者的躯干重心。38例无脊髓神经症状体征、双髋活动良好的AS胸腰段后凸畸形患者纳入研究。比较AS矫形患者手术前后矢状面平衡距离（ sagittal vertical axis,SVA ）、C7距髋轴中心水平距离（ horizontal distance between hip axis and C7,HDHC ）、T5距髋轴中心水平距离（ horizontal distance between hip axis and T5,HDHT5）、T9距髋轴中水平距离（ horizontal distance between hip axis and T9,HDHT9）、肺门距髋轴中心水平距离（ horizontal distance between hip axis and hilus pulmonis,HDHH ）,将矫形前后测量值做统计描述及配对t检验。采用组内相关系数（ ICC ）衡量HDHH的观察者间信度（inter-observerreliability）及观察者内复测信度（intra-observerreliability）。结果矫形前SVA、HDHC、HDHT5、HDHT9、HDHH分别为21.1 cm、12.7 cm、3.5 cm、-3.8 cm、2.7 cm;矫形后分别为9.1 cm、4.2 cm、-2.1 cm、-5.6 cm、0.9 cm。SVA、HDHC、HDHT5、HDHT9矫形前后变化有统计学意义,HDHH 矫形前后变化无统计学意义。HDHH 的观察者间总体信度与观察者内总体复测信度分别为0.958、0.963（ P＜0.001）。结论矫形前后肺门垂线通常总是落在髋轴上,肺门可作为躯干重心的影像学标记应用于AS胸腰段后凸畸形的矫形设计。     

带呼吸囊管式背心抗荷服的热负荷测定

测定5名志愿被试者在34－35℃环境中着用带呼吸囊管式背心抗荷服时的加权平均皮肤温度（TS）、囊下局部皮肤温度、口腔温度等指标。结果表明，与不带呼吸囊组比较，管式背心加呼吸囊后除局部皮肤温度略有升高（1．2±0．1℃）外，其他各项指标均无显著差别（P＞0．05）。初步认为，在环境热负荷强度不大时，该装备不会引起飞行员显著热应激。     

Abnormal expression of Col X, PTHrP, TGF-β, bFGF, and VEGF in cartilage with Kashin–Beck disease

The purpose of the current study was to investigate the abnormal expression of Col X, PTHrP, TGF-β, bFGF, and VEGF in cartilage from patients with Kashin–Beck disease (KBD) to understand the pathogenesis of chondronecrosis in KBD. Articular cartilage and growth plate cartilage collected were divided into four groups: control children (8 samples, 5 cases), KBD children (19 samples, 9 cases), control adults (8 samples, 6 cases), and KBD adults (16 samples, 15 cases). The presence of PTHrP, TGF-β₁, bFGF, VEGF, and collagen X in articular cartilage and in growth plate cartilage was analyzed by immunohistochemistry. Articular cartilage and growth plate were each divided in three zones, and the rate of positive cells was counted by light microscope for cytoplasmic and pericellular staining. Results showed that (1) in KBD children, Col X expression was lower in the deep zone of growth plate cartilage than in normal children; in articular cartilage of KBD adults, however, collagen X expression was higher in the middle zone compared to the controls; (2) staining for bFGF, PTHrP, TGF-β₁, and VEGF in KBD adult patients was prominent in the chondrocyte clusters and the eroded surface of articular cartilage, and the percentage of chondrocyte staining was significantly higher than in control samples (t = 3.64–10.34, df = 12 for children and 19 for adults, P = 0.002–0.0001); and (3) the enhanced PTHrP, TGF-β₁, and VEGF staining in the deep and middle zone of KBD articular cartilage correlated with the high incidence of chondronecrosis in the middle zone (48.5% ± 10.2%) and deep zone (70.6% ± 27.0%) of adult KBD cartilage. In conclusion, Col X expression was reduced in areas of chondrocyte necrosis in the deep zone of KBD articular cartilage, indicating changes in terminal chondrocyte differentiation. PTHrP, TGF-β₁, and VEGF expression was significantly altered and indicated degenerative changes in KBD cartilage, which initially resemble those occurring in osteoarthritis, but lead eventually to chondronecrosis, an event not observed in osteoarthritis.     

rhGM-CSF联合纳米银外用对深Ⅱ度烫伤创面细菌变化的影响

目的研究外用重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)联和纳米银敷料外用对深Ⅱ度烫伤创面愈合过程细菌变化的影响。方法用Wistar大鼠建立深Ⅱ度烫伤模型,分为A、B、C、D 4组,A组给予凡士林纱布覆盖,B组给予纳米银敷料覆盖,C组给予rhGM-CSF涂抹创面,D组给予rhGM-CSF+纳米银敷料覆盖,伤后第1,4,7,10,14,21天,观察创面炎症反应情况及愈合情况,进行细菌培养并计数。结果炎症反应,A组B组C组D组;4组均在第10天出现明显的血管化和上皮化;创面愈合率,D组C组B组A组;A组、B组/C组、D组分别于伤后第4,10,14天出现细菌生长,D组与A组、B组、C组相比,B组/C组与A组相比,细菌生长数量少,并且第10,14,21天比较均有显著性差异(P均0.05)。结论 GM-CSF和纳米银联合外用,可加速深Ⅱ度烫伤创面愈合,减少细菌生长,减轻炎症反应,并且优于rhGM-CSF、纳米银单独应用。     

大鼠心肌缺血再灌注后心肌梗死面积和无复流面积的动态变化及其相关性的初步研究

目的 研究大鼠心肌缺血再灌注后无复流现象中心肌梗死面积和无复流面积的动态变化及其相关性。方法 采用健康雄性Wistar大鼠,分两批进行实验。第1批实验,固定再灌注时间为2 h,将大鼠随机分为4组:①缺血2 h组,②缺血3 h组,③缺血4 h组,④假手术组。第2批实验,固定缺血时间为4 h,大鼠随机分为6组:①再灌注2 h组,②再灌注4 h组,③再灌注8 h组,④再灌注24 h组,⑤再灌注3 d组,⑥假手术组。缺血再灌注组结扎冠状动脉左前降支一定时间后再灌注,假手术组只穿线不结扎。分别用硫磺素S和硝基四氮唑蓝(N-BT)染色评估心肌无复流面积和梗死面积。结果 所有缺血再灌注组均出现明显的心肌无复流现象。心肌梗死面积和无复流面积随缺血时间的延长而扩大,缺血2 h,即可见明显的心肌无复流现象,无复流区与血液灌注区界限明确;缺血3 h,可见明显心肌梗死,但界限不清;缺血4 h梗死区与未梗死区界限清晰。再灌注8 h内,心肌梗死面积和无复流面积随再灌注时间的延长明显扩大、延展,无复流面积略大于梗死面积;再灌注8 h后,心肌梗死的扩展速度进入平台期,而无复流面积则呈逐渐减少趋势;再灌注3 d后,心肌梗死和无复流区域基本重合,二者面积趋于一致。结论 在本研究选定的缺血时间和再灌注时间的条件下,心肌缺血再灌注模型大鼠的心肌无复流发生率是100 %,可作为心肌无复流模型使用。心肌梗死面积和无复流面积随缺血时间和再灌注时间的延长而扩大,并均在再灌注8 h达峰值;此后梗死面积基本固定,而无复流现象出现部分恢复和改善。     

Differentiation of bel-7402 human hepatocarcinoma cells induced by aqueous extracts of fresh gecko (AG) and its anti-tumor activity in vivo

Ethnopharmacological relevance

Gecko, a kind of reptile, has been widely used as a traditional Chinese medicine to treat various diseases including cancer in China for thousands of years. The aim of this study was to investigate the anti-tumor effect of AG (aqueous extracts of fresh gecko) on human hepatocellular carcinoma cell Bel-7402 in vitro and mouse H22 hepatocellular in vivo. Further to underlie the molecular mechanism of AG inducing the differentiation of Bel-7402 cells.

Materials and methods

AG was obtained by water extracting method and qualitatively analyzed through High Performance Liquid Chromatography. The total protein concentration of AG was measured by BCA (bicinchoninic acid disodium) assay. The anti-tumor activities in vivo were analyzed through H22 (mouse hepatocellular carcinoma cell line H22) tumor xenografts mice. The cytotoxic activity of AG on Bel-7402 cells was evaluated by MTT assays. AFP (alpha fetoprotein) was detected by radioimmunoassay. ALB (albumin), ALP (alkaline phosphatase) and γ-GT (γ-glutamyl transpeptidase) were detected by biochemical methods with commercial kits. While morphological changes were observed through an inverted microscope. Moreover, the expression level of the proteins involved in MAPK (mitogen-activated protein kinase) signal pathway which was closely related to cellular differentiation was assessed by Western blot.

Results

AG showed obviously anti-tumor activity in vivo and anti-proliferative activity on Bel-7402 cells in vitro both dose-dependently. The number of clones of Bel-7402 cells treated with AG reduced and the cells were displaying differentiation state such as relatively bigger size and dispersed growth. The biochemical function markers of the cells were significantly changed after being treated with AG. The data showed that AFP secretion of the cells decreased 42.5%, ALB secretion increased 58.9%, the activity of ALP and γ-GT markedly decreased 67.0% and 48.5% separately when the concentration of AG was 10 μl/ml, and those effects were all in a dose-dependent manner. The major original and phosphorylated signal proteins (ERK1/2 (extracellular sigal-regualted kinase 1/2), P38 (p38 MAPK) and JNK1/2 (c-Jun N-terminal kinase 1/2)) involved in MAPK signal pathway were measured and the results showed that AG activated the ERK1/2 of Bel-7402 cells.

Conclusions

AG has anti-tumor activity in vivo and inhibits Bel-7402 cell proliferation in vitro through inducing cell differentiation, and the mechanism involves the activation of ERK1/2.     

Steroid receptor co-activator-3 promotes osteosarcoma progression through up-regulation of FoxM1

Increasing evidence suggests that the three homologous members of steroid receptor co-activator (SRC) family (SRC-1, SRC-2, and SRC-3) play key roles in enhancing cell proliferation in various human cancers, such as breast, prostate, and hepatocellular carcinoma. However, the function of SRC-3 in osteosarcoma remains largely unexplored. In the current study, we found that SRC-3, but not SRC-1 and SRC-2, was dramatically up-regulated in human osteosarcoma tissues, compared with adjacent normal tissues. To explore the functions of SRC-3 in osteosarcoma, in vitro studies were performed in MG63 and U2OS cells. SRC-3 overexpression promoted osteosarcoma cell proliferation, whereas knockdown of SRC-3 inhibits its proliferation. In support of these findings, we further demonstrated that SRC-3 up-regulated FoxM1 expression through co-activation of C/EBPγ. Together our results show that SRC-3 drives osteosarcoma progression and imply it as a therapeutic target to abrogate osteosarcoma.     

3678例已婚女性宫颈细胞HPV感染基因谱的研究

目的人乳头瘤病毒(HPV)感染是宫颈上皮内瘤变及宫颈癌的主要致病因子。该文旨在探讨南京地区已婚女性宫颈细胞中23种人乳头瘤病毒感染的基因谱分布情况及其临床意义。方法从南京地区扬子石化集团公司3678例已婚女性职工的宫颈上皮细胞标本中提取23种HPVDNA,采用基因扩增结合基因芯片技术对其宫颈细胞进行23种HPV基因型的检测,并对其受检者进行相关资料分析。结果 3678例已婚女性宫颈上皮细胞标本中检出HPV感染者397例,总的HPV感染率为10.79%(397/3678),其中单一型别的阳性检出率为9.11%(335/3678);单一型别的感染中HPV43型为80例,其阳性检出率为2.18%(80/3678),是最主要的感染型别;其次为HPV16型35例、58型33例,其阳性检出率分别为0.95%(35/3678)、0.90%(33/3678);混合型HPV感染62例,其阳性检出率为1.69%(62/3678);其中HPV43型+X型12例及43型+X型+X型10例、43+X+X+X型1例,占混合型感染的37.10%(23/62),是混合型感染的主要型别;其次是HPV16型的二和三重混合型16例,占混合型感染的25.81%(16/62)。结论 HPV43、16、58型单一型别及43型和16型的混合型是感染南京地区已婚女性宫颈细胞的主要基因型,基因扩增结合基因芯片检测技术可适用于宫颈细胞标本,一次可检测23种HPV感染的基因型,其特异性强,敏感性高,对我国已婚女性宫颈HPV感染基因型的分子流行病学的调查研究具有重要的意义。