宫颈癌术野血经辐射后回输的临床前实验研究

目的　了解有效去除血中污染的宫颈癌细胞所需的辐射剂量。方法　对人类宫颈癌HeLa系细胞给予不同剂量梯度的γ线辐射,测定细胞生长抑制率及克隆形成抑制率,从而确定完全抑制HeLa细胞增生的最低有效辐射剂量。为了模拟临床实际情况,在体外将HeLa细胞与洗涤红细胞混合后给予有效剂量辐射,经密度梯度分离HeLa细胞并接种培养,用细胞集落形成检测其增生能力。取40份妇科恶性肿瘤病人的静脉全血,分别测定其在无辐射及予以有效剂量辐射后红细胞携氧功能指标2,3 DPG值。结果　12 Gy的辐射剂量能够完全抑制肿瘤细胞的增生(细胞生长抑制率及克隆形成抑制率均为100%,P<0.01),且不影响红细胞的携氧功能(不辐射与辐射后的2,3 DPG值无显著性差异,P>0.05)。结论　12 Gy的辐射剂量是彻底去除血中污染的宫颈癌细胞的最低有效辐射剂量,且不影响红细胞的携氧功能;为在宫颈癌手术中回收式自体输血(IAT)的临床应用提供了实验依据。     

ABT737通过延迟宫颈癌细胞放射后DNA损伤修复及诱导凋亡而增敏放疗

【目的】 探讨类似BH-3的小分子物质ABT737诱导增敏放疗的作用及其分子机制?【方法】噻唑蓝法(MTT)检测不同浓度ABT737对HeLa细胞的生长抑制作用;细胞克隆形成实验检测ABT737联合放疗对放疗的增敏作用;免疫荧光法检测γ-H2AX观察DNA损伤修复情况;流式细胞术检测细胞凋亡;免疫印迹法检测Caspase-3?PARP的表达观察凋亡?【结果】与对照组相比,ABT737能显著抑制宫颈癌HeLa细胞的增殖(P < 0.05),并呈浓度依赖性,其IC50为15.7 μmol/L?体外培养克隆形成实验结果显示放疗+ABT737不同用药时间组的DEF值均大于1,放疗+8 μmol/L ABT737持续用药组为1.88,放疗+12 μmol/L ABT737用药72 h组为1.13,细胞存活分数(SF值)持续用药组为0.84,用药72 h组SF值为0.82;免疫荧光结果显示ABT737联合放疗处理HeLa细胞1 h后,放射线导致的γ-H2AX聚焦点数量及有γ-H2AX聚焦点生成的细胞数量均明显增加,上述处理24 h后,单纯放疗组γ-H2AX焦点消失,而联用ABT737处理组仍可观察到γ-H2AX焦点聚集?流式细胞术结果显示,单纯放疗组早期凋亡率(Annexin V+,PI-)为23.3%,ABT737联合放疗可以明显提高放射线诱导的细胞凋亡,早期凋亡率最高达50.3%?免疫印迹结果显示10 ?滋mol/L ABT737与2 Gy放射联合作用于Hela细胞后,凋亡蛋白cleaved Caspase-3与cleaved PARP的表达较单纯放疗组增加?【结论】 ABT737对宫颈癌Hela细胞具有放疗增敏作用,其机制与ABT737可延迟宫颈癌细胞放疗后DNA损伤修复及诱导凋亡有关?     

hTERT启动子驱动的TRAIL联合放化疗对HeLa裸鼠移植瘤生长抑制的作用

目的:探讨hTERT启动子驱动的TRAIL联合放化疗对宫颈癌HeLa裸鼠皮下移植瘤生长抑制的作用. 方法:将含有hTERT启动子的TRAIL基因载体转染宫颈癌细胞HeLa,检测稳定转染细胞中GFP/TRAIL的表达;实验裸鼠分为基因组,化疗组,放疗组,综合组(基因 放化疗),对照组,每组5只,未转染HeLa组为对照组,不作任何处理;将稳定转染基因载体及未转染的HeLa细胞种植于裸鼠背部皮下,联合放化疗,来观察移植瘤成瘤情况,移植瘤的体积、质量以及质量抑制率的变化. 结果:转染hTERT-TRAIL细胞中GFP/TRAIL(绿色荧光蛋白)表达率高于对照组,有统计学意义 (P＜0.01);各组裸鼠在接种细胞第5～7日全部长出肿瘤小结节,结节出现的时间差异无统计学意义(P＞0.05);转染hTERT -TRAIL细胞与对照组HeLa细胞裸鼠的移植瘤质量相比差异有统计学意义(P＜0.05); hTERT -TRAIL细胞在裸鼠体内所形成的移植瘤生长较慢, 4 wk后肿瘤质量抑制率为43.08%,联合放化疗后肿瘤质量抑制率为61.63%,有统计学意义(P＜0.01). 结论:hTERT启动子驱动的TRAIL对人宫颈癌裸鼠移植瘤有明显的抑制作用,联合放化疗能抑制裸鼠移植瘤的生长. 基因治疗联合放化疗等多途径综合治疗为以后的肿瘤研究和临床治疗提供思路.     

羟基喜树碱对宫颈癌HeLa细胞的放射增益作用及影响因素的研究

目的研究羟基喜树碱(HCPT)对人宫颈癌HeLa细胞系的放射增益作用以及给药时序、药物浓度和作用时间对放射增益作用的影响。方法MTT法测定HCPT作用于HeLa细胞的IC50。应用克隆形成和单击多靶数字模型方法检测HCPT的放射增益作用及不同给药时序、不同药物浓度和不同作用时间的细胞存活率及放射增益比(SER)。结果HCPT对HeLa细胞的IC50为62.1μmol/L。取2.85μmol/L(IC20)HCPT联合射线照射作用HeLa细胞,先给药后照射组SER=1.187、给药中照射组SER=1.573、先照射后给药组SER=1.850;而用28.5μmol/L(10×IC20)HCPT作用HeLa细胞,先照射后给药组SER=1.897;HCPT作用HeLa细胞4 h内放射增益作用改变明显,其后延长作用时间对放射增益无明显影响。结论HCPT对HeLa细胞具有放疗增益作用,其增益作用有时序、浓度和时间依从性。     

4-羟苯基维胺脂对宫颈癌HeLa细胞系的放射增敏作用

目的探讨4-羟苯基维胺脂(4-HPR)对HeLa细胞生长抑制和凋亡的影响;研究小剂量4-HPR对HeLa细胞的放射增敏效应。方法用四甲基偶氮唑蓝比色法(MTT)观察4-HPR对HeLa细胞的生长抑制情况;电镜观察4-HPR、放疗处理后HeLa细胞超微结构的改变;流式细胞仪检测单用4-HPR、γ-射线以及2者联合使用时HeLa细胞凋亡率和细胞周期变化。结果4-HPR和2 Gy60Co照射单独作用均可引起HeLa细胞超微结构改变,发生早期凋亡,4-HPR低剂量组(1μmol.L-1,2μmol.L-1)、单纯放射组与无药对照组凋亡率比较无统计学差异(P>0.05);4-HPR 4μmol.L-1组与无药对照组有显著性差异(P<0.01)。2 Gy60Co照射和4-HPR(2μmol.L-1,4μmol.L-1)联合使用细胞凋亡率较单纯放射组凋亡率有显著性差异(P<0.05,P<0.01);4-HPR作用后,HeLa细胞周期发生变化,表现为G1期减少,S期增加。结论4-HPR可诱导肿瘤细胞凋亡,小剂量4-HPR可增加HeLa细胞对放疗的敏感性。     

苦参碱抑制宫颈癌HeLa细胞增殖作用及机制

目的探讨苦参碱对人宫颈癌HeLa细胞体外增殖的抑制作用及其分子机制。方法以不同浓度(0.5、1.0、1.5、2.0g/L)的苦参碱分别处理HeLa细胞24、48、72h后,应用MTT法检测细胞增殖抑制率,WesternBlot方法检测培养48h细胞中真核细胞翻译起始因子4E(eIF4E)、4E结合蛋白1(4E-BP1)蛋白表达情况;1.0g/L的苦参碱作用HeLa细胞1、3、6、12h后,Western Blot方法测其eIF4E、4E-BP1蛋白磷酸化水平。结果苦参碱明显抑制HeLa细胞增殖,且呈时间和剂量依赖性:在相同时间内,随着苦参碱浓度的增加,HeLa细胞的增殖抑制率明显升高(F=235.035,P<0.05);在同一浓度时,随着苦参碱作用时间的延长,HeLa细胞的增殖抑制率也明显升高(F=320.207,P<0.05);2.0g/L苦参碱作用72h对HeLa细胞增殖抑制率最大,为(65.30±2.17)%。不同浓度苦参碱处理HeLa细胞48h后,eIF4E、4E-BP1蛋白的表达均无明显变化(F=0.171、0.932,P>0.05)。1.0g/L苦参碱作用于HeLa细胞不同时间后,细胞中eIF4E、4E-BP1蛋白磷酸化水平均降低,且呈时间依赖性(F=43.48、107.23,P<0.01)。结论苦参碱可以明显抑制体外培养宫颈癌HeLa细胞的增殖,其作用可能是通过抑制eIF4E以及4E-BP1的磷酸化,从而抑制蛋白翻译来实现的。     

菠萝蛋白酶对宫颈癌细胞HeLa增殖及凋亡的影响

目的:探讨菠萝蛋白酶对人宫颈癌细胞HeLa增殖和凋亡的影响。方法:将人宫颈癌HeLa细胞用不同剂量的菠萝蛋白酶处理48h后,光学显微镜下观察其形态学变化;采用MTT法检测不同剂量菠萝蛋白酶作用不同时间(24h、48h、72h)后对HeLa细胞增殖的影响;然后采用FITC-annexin V/PI双染色法检测不同剂量处理组HeLa细胞的凋亡情况。结果:菠萝蛋白酶对HeLa细胞的生长抑制作用呈明显的时间、剂量依赖性;且随着剂量增加,能够诱导HeLa细胞发生凋亡,与对照组相比具有显著的统计学意义(P<0.05)。结论:菠萝蛋白酶能明显抑制宫颈癌HeLa细胞的增殖,可能是通过诱导细胞凋亡的发生,从而发挥其抗肿瘤效应。     

加热联合紫杉醇对人喉癌细胞Hep-2体外增殖抑制及凋亡的影响

目的 探讨加热(hyperthermia,HT)联合紫杉醇(Taxol)对人喉癌细胞系(Hep-2细胞)体外增殖抑制及凋亡的影响.方法 将Hep-2细胞分为实验组与对照组,实验组用不同浓度(0.1、1.0及10.0μmol/L)的Taxol预处理后联合热疗(39、41及43℃1h)处理不同时间(24、48及72h)后,采用Wright-Gimsa染色法观察Hep-2细胞凋亡的形态学变化;以四甲基偶氮唑盐(MTT)还原试验检测细胞活力,以细胞增殖率作为细胞损伤指标,流式细胞术检测细胞凋亡发生率.结果 加热41℃组细胞增殖率与其他不同温度同剂量组比较均显著降低(P＜0.01);39℃组与对照组37℃组比较差异无统计学意义(P＞0.05);热疗联合Taxol组处理细胞48h后,细胞出现凋亡形态学改变.与热疗和Taxol单药组相比,热疗联合Taxol组对细胞的抑制率显著增强(P＜0.01),其作用呈时间和剂量依赖性;热疗联合Taxol组诱导细胞凋亡率与单药组及单热疗组相比较均增高(P＜0.05);而单药紫杉醇组与单独热疗组之间比较差异无统计学意义(P＞0.05).结论 热疗和Taxol单药体外均可抑制Hep-2细胞增殖并诱导其调亡.热疗和Taxol联合应用对Hep-2细胞体外增殖抑制其诱导凋亡作用显著增强;诱导凋亡可能是细胞增殖抑制的作用机制之一.     

72例宫颈癌热放疗联合治疗疗效分析

目的:评价放疗联合局部热疗治疗宫颈癌的临床疗效及毒副反应。方法:2001年6月至2003年12月收治的72例宫颈癌患者随机分为两组:放疗+热疗综合治疗(综合治疗组)36例,单纯放疗组(对照组)36例。比较两组治疗后的近期疗效、临床受益疗效、生存期及毒副反应。结果:综合组、对照组有效率分别为91.7%和72.3%,综合治疗组的临床获益疗效明显较对照组高,两组1a、3a生存率比较P>0.05,差异无显著性5a生存率,P<0.05,差异有显著性。两组毒副反应比较P>0.05无显著意义。结论:放疗+热疗联合治疗宫颈癌能显著提高宫颈癌的近期疗效、生存期。而其两组的毒副反应比较无明显差异。     

生理性深层海水联合热疗治疗肝细胞癌的体外抑瘤研究

目的：探讨生理性深层海水（PDSW）联合热疗对肝细胞癌的体外抑瘤作用。方法将取自我国海南省海域的深层海水进行制备,制成PDSW,检测所含有的部分元素。体外培养的正常肝细胞和人肝癌QGY‐7703细胞被随机分为PDSW组和生理盐水组,PDSW组加入PDSW,生理盐水组给予等量生理盐水,24h后,每天分别接受40℃热疗6h和43℃热疗1h,在热疗后的24、48、72h,用MTT法检测热疗结合PDSW对正常肝细胞及人肝癌QGY‐7703细胞的抑制率。同时检测PDSW及生理盐水在40℃6h连续10d状态下对人肝癌QGY‐7703细胞克隆形成率的影响。结果MTT检测结果显示肿瘤抑制率在两组均呈时间和浓度依赖性。PDSW组的肿瘤抑制率明显较生理盐水组高,差异有统计学意义（P＜0．05）。PDSW组对正常肝细胞的生长抑制率明显低于生理盐水组。此外,PDSW组的肿瘤细胞克隆形成率较生理盐水组低,差异有统计学意义（P＜0．05）。结论PDSW能够提高正常肝细胞对热的耐受性。当联合热疗时,可明显抑制人肝癌QGY‐7703细胞生长。     

UF-100检测北京地区正常人群尿液有形成分的调查

目的应用UF-100检测北京地区健康人群尿液的有形成分,为临床建立尿液有形成分生物参考范围。方法应用UF-100尿液分析仪检测6587例健康体检者(其中男性3243人,女性3344人,年龄为1~92岁)随机中段尿液红细胞(RBC)、白细胞(WBC)、上皮细胞(EC)及管型数。结果尿液有形成分红细胞(12岁以下组除外)、白细胞、上皮细胞检测结果性别之间差异有统计学意义(P<0.05)。白细胞检测结果男女13岁以上各年龄组之间差异均无统计学意义(P>0.05),而与12岁以下年龄组差异有统计学意义(P<0.05);红细胞检测结果男性12岁以下与19岁以上各年龄组之间差异无统计学意义(P>0.05),13~18岁年龄组与其他各年龄组差异均有统计学意义(P<0.05)。女性13岁以上各年龄组之间差异无统计学意义(P>0.05)而与12岁以下年龄组差异有统计学意义。上皮细胞检测结果男性各年龄组之间差异无统计学意义(P>0.05),女性13~55岁年龄组之间差异无统计学意义(P>0.05),但与12岁以下及56岁以上年龄组差异有统计学意义(P<0.05)。管型检测结果显示性别、年龄组之间差异无统计学意义(P>0.05)。结论尿液RBC、WBC、EC与性别、年龄有关,在临床应用时,应注意男女性别之间、年龄之间生物参考范围不同。     

异位妊娠发病因素十年变迁

目的探讨不同时期异位妊娠发病因素的差异,以探索切实可行的干预措施。方法对首都医科大学附属北京妇产医院1997年10月～1998年9月期间(前期组)186例和2007年10月～2008年9月期间(后期组)382例异位妊娠患者发病的相关因素进行回顾性分析,包括年龄、婚姻状况、孕产史、职业分布、既往病史和治疗情况等。结果 2组患者平均年龄差异无统计学意义,后期组20岁以下构成比由10年前的1.1%上升至4.5%(P=0.006);未婚女性由1997年的21.5%上升至2007年的41.4%(P=0.000);未育女性由1997年的45.7%上升至2007年的66.8%(P=0.000);2组间孕、产次差异无统计学意义,但后期组流(引)产次数明显高于前期组(P=0.030);职业分布:工人、农民的构成比明显下降,干部、职员和学生、其他职业的构成比逐年上升(P=0.000);前期组患者宫内节育器病史明显多于后期组(P=0.000),而生殖道感染史和其他腹部手术史明显少于后期组(P=0.002,P=0.044)。后期组中阴道灌洗史、宫颈治疗史及吸烟史均较前期组明显增高(P=0.000,P=0.009,P=0.002)。结论 10年来异位妊娠发病相关因素发生了变化,应采取相应的措施,以降低异位妊娠的发生。     

Effects of cloned tumstatin-related and angiogenesis-inhibitory peptides on proliferation and apoptosis of endothelial cells

Background Tumstatin is a recently developed endogenous vascular endothelial growth inhibitor that can be applied as an anti-angiogenesis and antineoplastic agent. The study aimed to design and synthesize the small molecular angiogenesis inhibition-related peptide （peptide 21）, to replicate the structural and functional features of the active zone of angiogenesis inhibition using tumstatin and to prove that synthesized peptide 21 has a similar activity： specifically inhibiting tumor angiogenesis like tumstatin. Methods Peptide 21 was designed and synthesized using biological engineering technology. To determine its biological action, the human umbilical vein endothelial cell line ECV304, the human ovarian cancer cell line SKOV-3 and the mouse embryo-derived NIH3T3 fibroblasts were used in in vitro experiments to determine the effect of peptide 21 on proliferation of the three cell lines using the MTT test and growth curves. Transmission electron microscopy （TEM） and flow cytometry （FCM） were applied to analyze the peptide 21-induced apoptosis of the three cell lines qualitatively and quantitatively. In animal experiments, tumor models in nude mice subcutaneously grafted with SKOV-3 were used to observe the effects of peptide 21 on tumor weight, size and microvessel density （MVD）. To initially investigate the role of peptide 21, the effect of peptide 21 on the expression of vascular endothelial growth factors （VEGFs） by tumor tissue was semi-quantitatively analyzed. Results The in vitro Ml-r test and growth curves all indicated that cloned peptide 21 could specifically inhibit ECV304 proliferation in a dose-dependent manner （P 〈0.01）; TEM and FCM showed that peptide 21 could specifically induce ECV304 apoptosis （P 〈0.01）. Results of in vivo experiments showed that tumors in the peptide 21 group grew more slowly. The weight and size of the tumors after 21 days of treatment were smaller than those in the control group （P 〈0.05）, with a mean tumor inhibition rate of 67.86%; MVD     

766例宫颈病变临床诊断方法的分析

目的评价宫颈液基细胞学、阴道镜和镜下宫颈组织病理学检查结果对宫颈病变的应用价值,以便早期发现和诊断宫颈上皮内瘤变和宫颈癌。方法北京市垂杨柳医院妇科门诊在2002年6月至2005年2月,行宫颈基薄层细胞学(TCT)初筛6 201例,对其中442例阳性和324例阴性但临床可疑宫颈病变共计766例,进一步行阴道镜及镜下取活检病理检查,以病理组织学为金标准,对诊断结果进行比较分析。结果TCT初筛6 201例,阳性病变442例,发病率7.13%,其中高度鳞状上皮内病变(HSIL)21例,低度鳞状上皮内病变(LSIL)82例,宫颈癌(CC)4例,意义不明的不典型腺细胞(AGUS)335例。阴道镜和病理诊断HSIL分别为34和36例,LSIL分别为143和153例,CC均为5例,炎症分别为584和572例。阴道镜对≥LSIL和≥HSIL诊断的Kappa值分别为0.887和0.868,均高于TCT对≥LSIL和≥HSIL诊断的Kappa值(0.615和0.716)。TCT和阴道镜诊断对≥LSIL以上病变的敏感性为53.1%和88.7%(P<0.01),特异性为99.3%和98.3%(P>0.05),符合率为87.6%和95.9%(P<0.01),漏诊率为46.9%和11.3%(P<0.01),阴性预测值为86.2%和96.2%(P<0.01),阳性预测值为96.3%和94.5%(P>0.05),误诊率为0.7%和1.7%(P>0.05)。335例ASCUS/AGUS病理诊断为宫颈炎症282例,宫颈上皮内瘤变(CIN)Ⅰ48例,CINⅡ3例,CINⅢ2例。结论液基细胞学可用于宫颈病变的初筛,液基细胞学检查联合阴道镜下活组织检查,可提高宫颈病变的检出率及准确率,可将二者联合用于宫颈上皮内瘤变的诊断。对ASCUS患者需进一步检查。     

子宫内膜癌治疗方法的评价

目的评价子宫内膜癌不同治疗方法的疗效。方法对1960 2001年间北京妇产医院收治的867例子宫内膜癌的临床及病理资料进行回顾性分析。按治疗方法归纳为3组:术前腔内半量放疗+手术组(HR+S组)、手术组(S组)、单纯放疗组(R组)。结果临床Ⅰ期(570例)HR+S组、S组、R组5年生存率分别为93.7%、90.7%和57.8%;临床Ⅱ期(134例)3组的5年生存率分别为88.3%、64.3%和25.1%。Ⅰ期与Ⅱ期5年生存率HR+S组与S组之间比较,差异均无统计学意义(P>0.05),与R组相比差异均有统计学意义(P<0.01)。结论对于临床Ⅰ、Ⅱ期子宫内膜癌,综合治疗明显好于单纯放疗;术前腔内半量放疗再行全子宫双附件切除术与手术(筋膜外次广泛全子宫双附件切除),术后高危型均补充放疗,2种治疗方法疗效无差异。     

基质金属蛋白酶-9、血管内皮生长因子与造釉细胞型颅咽管瘤侵袭、复发的相关性研究

目的探讨基质金属蛋白酶-9(matrix metalloproteinase9,MMP-9)和血管内皮生长因子(vascular endothelial growth factor,VEGF)在造釉细胞型颅咽管瘤中的表达及其与肿瘤侵袭、复发的相关性。方法应用免疫组化法对40例造釉细胞型颅咽管瘤标本(原发组24例,复发组16例)中MMP-9和VEGF的表达进行检测。结果 MMP-9在复发组造釉细胞型颅咽管瘤中的总阳性表达率93.7%,明显高于原发组总阳性率41.7%(P<0.05);VEGF在复发组造釉细胞型颅咽管瘤中的总阳性表达率87.5%,明显高于原发组总阳性率45.8%(P<0.05)。复发组釉质型颅咽管瘤中MMP-9与VEGF表达呈正相关(P<0.05),原发组造釉细胞型颅咽管瘤中MMP-9与VEGF表达无明显相关性。结论 MMP-9和VEGF可能在造釉细胞型颅咽管瘤侵袭生长和肿瘤复发过程中扮演着重要的角色,并可能存在协同作用。MMP-9和VEGF可能成为评价造釉细胞型颅咽管瘤侵袭性生长和复发倾向的重要参考指标。     

Cognitive-behavior therapy, sertraline, and their combination for children and adolescents with obsessive-compulsive disorder: the Pediatric OCD Treatment Study (POTS) randomized controlled trial

Context  The empirical literature on treatment of obsessive-compulsive disorder (OCD) in children and adolescents supports the efficacy of short-term OCD-specific cognitive-behavior therapy (CBT) or medical management with selective serotonin reuptake inhibitors. However, little is known about their relative and combined efficacy. Objective  To evaluate the efficacy of CBT alone and medical management with the selective serotonin reuptake inhibitor sertraline alone, or CBT and sertraline combined, as initial treatment for children and adolescents with OCD. Design, Setting, and Participants  The Pediatric OCD Treatment Study, a balanced, masked randomized controlled trial conducted in 3 academic centers in the United States and enrolling a volunteer outpatient sample of 112 patients aged 7 through 17 years with a primary Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition diagnosis of OCD and a Children’s Yale-Brown Obsessive-Compulsive Scale (CY-BOCS) score of 16 or higher. Patients were recruited between September 1997 and December 2002. Interventions  Participants were randomly assigned to receive CBT alone, sertraline alone, combined CBT and sertraline, or pill placebo for 12 weeks. Main Outcome Measures  Change in CY-BOCS score over 12 weeks as rated by an independent evaluator masked to treatment status; rate of clinical remission defined as a CY-BOCS score less than or equal to 10. Results  Ninety-seven of 112 patients (87%) completed the full 12 weeks of treatment. Intent-to-treat random regression analyses indicated a statistically significant advantage for CBT alone (P = .003), sertraline alone (P = .007), and combined treatment (P = .001) compared with placebo. Combined treatment also proved superior to CBT alone (P = .008) and to sertraline alone (P = .006), which did not differ from each other. Site differences emerged for CBT and sertraline but not for combined treatment, suggesting that combined treatment is less susceptible to setting-specific variations. The rate of clinical remission for combined treatment was 53.6% (95% confidence interval [CI], 36%-70%); for CBT alone, 39.3% (95% CI, 24%-58%); for sertraline alone, 21.4% (95% CI, 10%-40%); and for placebo, 3.6% (95% CI, 0%-19%). The remission rate for combined treatment did not differ from that for CBT alone (P = .42) but did differ from sertraline alone (P = .03) and from placebo (P<.001). CBT alone did not differ from sertraline alone (P = .24) but did differ from placebo (P = .002), whereas sertraline alone did not (P = .10). The 3 active treatments proved acceptable and well tolerated, with no evidence of treatment-emergent harm to self or to others. Conclusion  Children and adolescents with OCD should begin treatment with the combination of CBT plus a selective serotonin reuptake inhibitor or CBT alone.      

A novel and feasible way to cultivate and purify endothelial progenitor cells from bone marrow of children with congenital heart diseases

Background  Endothelial progenitor cells (EPCs) are used in vascular tissue engineering and clinic therapy. Some investigators get EPCs from the peripheral blood for clinic treatment, but the number of EPCs is seldom enough. We have developed the cultivation and purification of EPCs from the bone marrow of children with congenital heart disease, to provide enough seed cells for a small calibre vascular tissue engineering study.

Methods  The 0.5-ml of bone marrow was separated from the sternum bone, and 5-ml of peripheral blood was collected from children with congenital heart diseases who had undergone open thoracic surgery. CD34+ and CD34+/VEGFR+ cells in the bone marrow and peripheral blood were quantified by flow cytometry. CD34+/VEGFR+ cells were defined as EPCs. Mononuclear cells in the bone marrow were isolated by Ficoll^® density gradient centrifugation and cultured by the EndoCult Liquid Medium Kit™. Colony forming endothelial cells was detected. Immunohistochemistry staining for Dil-ac-LDL and FITC-UEA-1 confirmed the endothelial lineage of these cells.

Results  CD34+ and CD34+/VEGFR+ cells in peripheral blood were (0.07±0.05)% and (0.05±0.02)%, respectively. The number of CD34+ and CD34+/VEGFR+ cells in bone marrow were significantly higher than in blood, (4.41±1.47)% and (0.98±0.65)%, respectively (P <0.0001). Many colony forming units formed in the culture. These cells also expressed high levels of Dil-ac-LDL and FITC-UEA-1.

Conclusion  This is a novel and feasible approach that can cultivate and purify EPCs from the bone marrow of children with congenital heart disease, and provide seed cells for small calibre vascular tissue engineering.

     

Significance of Aspergillus spp. isolation from lower respiratory tract samples for the diagnosis and prognosis of invasive pulmonary aspergillosis in chronic obstructive pulmonary disease

Background  Chronic obstructive pulmonary diseases (COPD) is an emerging population at risk for invasive infection of Aspergillus. Isolation of Aspergillus from lower respiratory tract (LRT) samples is important for the diagnosis of invasive pulmonary aspergillosis (IPA). The purpose of this study was to investigate the value of Aspergillus isolation from LRT samples for the diagnosis and prognosis of IPA in COPD population.

Methods  Clinical record with Aspergillus spp. isolation in COPD and immunocompromised patients was reviewed in a retrospective study. Patients were categorized and compared according to their severity of illness (admitted to general ward or ICU) and immunological function (COPD or immunocompromised).

Results  Multivariate statistical analysis showed that, combined with Aspergillus spp. isolation, APACHE II scores >18, high cumulative doses of corticosteroids (>350 mg prednisone or equivalent dose) and more than four kinds of broad-spectrum antibiotics received in hospital may be predictors of IPA in COPD (OR=9.076, P=0.001; OR=4.073, P=0.026; OR=4.448, P=0.021, respectively). The incidence of IPA, overall mortality, mortality of patients with IPA and mortality of patients with Aspergillus spp. colonization were higher in COPD patients in ICU than in general ward, but were similar between COPD and immunocompromised patients.

Conclusions  Aspergillus spp. isolation from LRT in COPD may be of similar importance as in immunocompromised patients, and may indicate an increased diagnosis possibility of IPA and worse prognosis when these patients received corticosteroids, antibiotics, and need to admit to ICU. Aspergillus spp. isolation from LRT samples combined with certain risk factors may be useful in differentiating colonization from IPA and evaluating the prognosis of IPA in COPD patients.