Biomaterial-based platforms for cancer stem cell enrichment and study

Cancer stem cells (CSCs) are a relatively rare subpopulation of tumor cell with self-renewal and tumorigenesis capabilities. CSCs are associated with cancer recurrence, progression, and chemoradiotherapy resistance. Establishing a reliable platform for CSC enrichment and study is a prerequisite for understanding the characteristics of CSCs and discovering CSC-related therapeutic strategies. Certain strategies for CSC enrichment have been used in laboratory, particularly fluorescence-activated cell sorting (FACS) and mammosphere culture. However, these methods fail to recapitulate the in vivo chemical and physical conditions in tumors, thus potentially decreasing the malignancy of CSCs in culture and yielding unreliable research results. Accumulating research suggests the promise of a biomaterial-based three-dimensional (3D) strategy for CSC enrichment and study. This strategy has an advantage over conventional methods in simulating the tumor microenvironment, thus providing a more effective and predictive model for CSC laboratory research. In this review, we first briefly discuss the conventional methods for CSC enrichment and study. We then summarize the latest advances and challenges in biomaterial-based 3D CSC platforms. Design strategies for materials, morphology, and chemical and physical cues are highlighted to provide direction for the future construction of platforms for CSC enrichment and study.     

女性原发性骨质疏松性骨折患者生命质量影响因素分析

目的:评价女性原发性骨质疏松性骨折患者的生命质量并探讨其影响因素,为提高女性骨质疏松性骨折患者的生命质量提供理论依据。方法:采用横断面调查的方法,应用SF-36健康调查量表和自制一般情况问卷对79例女性原发性骨质疏松性骨折患者进行调查。结果:除精神健康外,生命质量各维度得分女性原发性骨质疏松性骨折患者均低于大于等于60岁四川省城市女性,且差异均有统计学意义(P<0.05)。单因素分析发现,不同年龄、婚姻状况及月经史间比较,生理健康总分各组间差异均有统计学意义(P<0.05);不同年龄、文化程度和骨折时间间比较,心理健康总分各组间差异均有统计学意义(P<0.05);不同年龄、吸烟、骨折次数和月经史间比较,生命质量总分各组间差异均有统计学意义(P<0.05)。多元线性回归分析发现,年龄和婚姻状况是女性原发性骨质疏松性骨折患者生理健康的独立影响因素,年龄和文化程度是心理健康的独立影响因素,年龄和骨折次数是生命质量总分的独立影响因素。结论:女性原发性骨质疏松性骨折患者的生命质量差,且年龄、婚姻状况、文化程度及骨折次数是其主要影响因素,应该采取针对性的预防干预措施提高患者的生命质量。     

Molecular characteristics of rifampin and isoniazid resistant Mycobacterium tuberculosis strains from Beijing, China

Background China is one of the high burden countries of Mycobacterium tuberculosis （TB） infection globally, with high incidence and mortality. We studied the molecular characteristics of rifampin （RIF） and isoniazid （INH） resistant Mycobacterium tuberculosis strains from Beijing, China, in order to find out the genetic marker for rapid detection of specific drug resistance. Methods Forty pansusceptible and 81 resistant strains of Mycobacterium tuberculosis isolated from Beijing, China during 2002-2005 were analyzed. The modified rifampin oligonucleotide （RIFO） assay based on reverse line blot hybridization was used to detect mutations in the 81 bp hot-spot region of rpoB gene, which is associated with RIF resistance. The INH resistance associated genes, regulatory region mab-inhA （-15C/T） and structural gene katG S315T were detected by reverse line blot hybridization and PCR-restriction fragment length polymorphism （RFLP） method respectively. All the strains were typed by spoligotying and the Beijing genotype was further subdivided by NTF locus analysis. The distribution of drug resistance associated mutations in the above genes was compared in these groups. Results Sixty-five （91.5%） of 71 RIF resistant and 52 （92.9%） of 56 multidrug-resistant （MDR, i.e. resistant to at least RIF and INH） strains were found to harbor mutations in the rpoB hot-spot region. No mutation was detected in RIF sensitive strains. The specificity and sensitivity of the modified RIFO assay were 100% and 91.5%, respectively, katG315 AGC〉ACC and inhA-15C〉T mutations were found in 40 （60.6%） and 10 （15.2%） of 66 INH resistant strains, respectively; 7.6% of INH-resistant strains had mutations in both of these genes. Therefore, a combined use of both katG315 and inhA-15 identified 68.2% of INH-resistant strains. The Beijing genotype accounted for 91.7% of total strains and was further subdivided into ＂modern＂ （76.6%） and ＂ancestral＂ （23.4%） group. There is no significant difference between ＂ancestral＂ and ＂modern＂ group in prevalance of drug resistance-associated gene mutations. Conclusions The hot-spot region of rpoB gene can be used as genetic marker for detection of RIF resistant strains; a combined use of both katG315 and inhA-15 can improve the detection rate of I NH resistant strains; the Beijing genotype is prevalent in Beijing, China; the modified RIFO assay can be a practical tool for rapid detection of RIF resistant and MDR isolates in the routine diagnostic work.     

应用明胶海绵栓塞法建立猪的心肌梗死模型

目的经心导管用明胶海绵栓子栓塞猪的冠状动脉左前降支(LAD),建立心肌梗死的大动物模型。方法对7头猪用明胶海绵经导管栓塞LAD第二对角支远端,致前室壁梗死。四周后行心脏超声、冠状动脉造影及组织学检查。结果手术死亡率为1/7。与对照组相比,心肌梗死(MI)组左室舒张未内径(LVEDd)增大由37.0mm±3.4mm增至50.8mm±6.1mm(P<0.01),射血分数(EF)由62.3%±2.9%下降为36.6%±2.1%(P<0.001);复查冠脉造影提示LAD栓塞仍存在。MI组的左室被动容积为81.4ml±4.3ml,梗死区面积/左室心肌面积为18.4%±1.6%,梗死区厚3.5mm±0.8mm。心肌组织马森染色显示梗塞交界区有大块纤维形成,梗塞区有片状纤维形成。结论这种模型接近临床病理生理过程,稳定可靠,重复性好,可作为研究心肌梗死的技术平台。     

重组转化生长因子β3基因对大鼠肝纤维化细胞模型胶原合成及沉积的影响

目的 观察重组转化生长因子β3(TGF-β3)基因对大鼠肝纤维化细胞模型胶原合成及沉积的影响.方法 (1)构建质粒pcDNA3.1(+)-TGF-β3和pcDNA3.1(+)-TGF-β1.(2)将pcDNA3.1(+)-TGF-β1转染大鼠肝星状细胞(HSC)-T6,经G418筛选建立稳定高表达TGF-β1的HSC-T6细胞阳性克隆.(3)pcDNA3.1(+)-TGF-B3转染HSC-T6阳性细胞克隆,荧光实时定量PCR法检测TGF-β3mRNA的表达;荧光实时定量PCR法及Western印迹法分别检测TGF-β3、Ⅰ型胶原、基质金属蛋白酶(MMP)-2、MMP-9及TIMP-1的mRNA和蛋白表达情况.结果 (1)pcDNA3.1(+)-TGF-β3和pcDNA3.1(+)-TGF-β3质粒均可转染HSC-T6细胞,以转染48 h时转染效率最高,达28.2%.(2)pcDNA3.1(+)-TGF-β3转染HSC-T6细胞阳性克隆后,与单纯阳性克隆组相比,TGF-β1mRNA表达无明显改变,而蛋白表达明显低(0.48±0.07 vs 0.66±0.14,P=0.023);Ⅰ型胶原mRNA及蛋白的表达明显低(mRNA:7.2±1.0 vs 13.7±0.4,P=0.010,蛋白:0.45±0.21 vs 0.82±0.13,P=0.041),MMP-2 mRNA及蛋白较低,但差异无统计学意义(均P>0.05),MMP-9 mRNA及蛋白表达明显多(均P<0.05),TIMP-1 mRNA及蛋白表达明显低(均P<0.05).结论 (1)重组TGF-β3真核表达载体构建正确,在转染阳性克隆细胞48 h后获得高效表达;(2)稳定高表达TGF-B1的HSC-T6细胞阳性克隆可作为肝纤维化细胞模型;(3)pcDNA3.1(+)-TGF-β3转染阳性克隆能减少胶原合成,并通过调节基质金属蛋白酶及其抑制剂的表达对胶原沉积起抑制作用.     

生脉注射液联合LMWH治疗CAHD不稳定型心绞痛临床研究

目的:探讨生脉注射液联合低分子量肝素钠(LMWH)治疗冠状动脉粥样硬化性心脏病(CAHD)不稳定心绞痛(UA)的临床疗效.方法:将70例UA患者随机分为治疗组(n=35)和对照组(n=35).两组均给予吸氧、卧床休息、调降脂、阿司匹林、β受体阻滞剂、血管紧张素转换酶抑制剂/血管紧张素受体阻滞剂(ACEI/ARB)、单硝酸异山梨酯及按需硝酸甘油片舌下含服等常规治疗,治疗组同时加用LMWH腹壁皮下注射,12小时1次,连用7d;生脉注射液静滴,每日1次,连用10天.结果:治疗组与对照组相比,治疗后心绞痛发作频率下降,心绞痛持续时间减少(P＜0.01).治疗组临床疗效及心电图疗效与对照组相比差异有统计学意义(P＜0.05).结论:生脉注射液联合LMWH治疗UA具有疗效显著、安全可靠、见效快、疗程短、毒副作用小等诸多优点.     

脑膜癌25例临床分析

目的 探讨脑膜癌的临床、影像学及脑脊液特点.方法 回顾性分析25例脑膜癌患者的临床、影像学及实验室检查资料.结果 25例患者中,男14例,女11例.头痛19例,恶心呕吐15例,复视7例,视力下降6例,听力下降5例,癫痫8例,智能减退6例,小便障碍6例.所有患者均行腰椎穿刺检查,脑脊液白细胞升高21例,蛋白升高17例,葡萄糖降低17例.25例患者均行MRI检查,脑膜强化征15例,脊膜强化征6例,脊髓内病变7例.结论 脑膜癌是恶性肿瘤颅内转移的特殊形式,脑脊液细胞学检查是诊断金标准,影像学显示脑膜结节样或混合型强化是明确诊断的重要依据.     

透明质酸修饰的尿酸酶多囊脂质体的体外特性及药效学过程

目的 研究透明质酸修饰的尿酸酶(UC)多囊脂质体(uricase in hyaluronic acid-uricase multivesicular liposomes,UHMVLs)的体外特性及其在大鼠体内的药效学.方法 采用复乳法制备UHMVLs并测定包封率及理化特性.取12只健康雄性SD大鼠,模型组采用次黄嘌呤和氧嗪酸钾建立高尿酸血症大鼠模型(n=3),UHMVLs组(n=3)和UC组(n=3)分别于建模后尾静脉注射UHMVLs和游离UC,并以正常组(n=3)为对照,测定大鼠血清中尿酸水平.结果 UHMVLs的平均包封率为(62.48±3.87)％ (n=3).UHMVLs和UC最适温度均为40℃,最适pH值分别为8.0和8.5.在同一温度(20～70℃)和pH(6.5～9.5)条件下,UHMVLs中UC的活性均高于游离UC(P＜0.05).除1、36、48 h外,其余各时间点UHMVLs降低高尿酸血症大鼠模型血清中尿酸水平的效果均较游离UC更显著(P＜0.05).结论 在相同条件下,UHMVLs不仅能提高UC的活性,还可以增强UC的稳定性;UHMVLs在大鼠体内降低血尿酸水平的能力优于游离UC.     

Inhibiting effect of antisense hTRT on telomerase activity of human liver cancer cell line SMMC-7721

Objective: To induce changes in biological character of human liver cancer cell line SMMC-7721 by blocking the expression of telomerase genes hTRT and to explore its value in cancer gene therapy. Methods: The vehicle for eukaryotic expression of antisense hTRT was constructed and then transfected into SMMC-7721 cells. The effects of antisense hTRT gene on telomerase activity, cancer cell growth and malignant phenotypes were analyzed. Results: The obtained transfectants that could express antisense hTRT gene stably showed marked decrease in telomerase activity;the shortening of telomere was obvious; cells presented contact growth inhibition; in nude mice transplantation, the rate of tumor induction dramatically decreased. Conclusion : Antisense hTRT gene expression can significantly inhibit telomerase activity of cancer cells and decrease malignant phenotypes in vitro and in vivo. Therefore, as a telomerase inhibitor, antisense hTRT gene may be a new pathway for cancer therapy.     

中国人群CCR5△32、CCR2-64I、SDF1-3’A基因多态性与HIV-1感染相关性的Meta分析

目的探讨中国人群CCR5△32、CCR2-64I、SDF1-3’A三种基因的多态性与HIV-1感染之间的关系。方法检索国内外有关中国人群CCR5△32、CCR2-64I、SDF1-3’A基因多态性与HIV感染相关性的病例对照研究文献并进行Meta分析。结果符合CCR5△32的研究共14个,累计HIV感染者病例1607例,正常对照组1632例。杂合子(wt/mt)、纯合子(mt/mt)和杂合子加纯合子(wt/mt+mt/mt)与野生基因型(wt/wt)相比合并的OR值(95%CI)分别为1.156(0.808,1.654)、0.997(0.198,5.022)、1.149(0.808,1.634)。符合CCR2-64I的研究共12个,累计HIV感染者病例1415例,正常对照组1239例。杂合子(wt/mt)、纯合子(mt/mt)和杂合子加纯合子(wt/mt+mt/mt)与野生基因型(wt/wt)相比合并的OR值(95%CI)分别为1.005(0.844,1.197)、1.191(0.808,1.754)、1.028(0.870,1.214)。符合SDFl-3’A的研究10个,累计HIV感染者病例1179例,正常对照组1003例。杂合子(wt/mt)、纯合子(mt/wt)和纯合子加杂合子(wt/mt+mt/mt)与野生基因型(wt/wt)相比合并的OR值(95%CI)分别为1.010(0.830,1.228)、1.188(0.860,1.643)、1.038(0.861,1.250)。结论中国人群CCR5△32、CCR2-64I、SDFl-3’A三种抗性基因与HIV-1感染之间无相关性,显示CCR5△32、CCR2-641、SDFl-3’A三种基因对中国人群HIV-1感染可能不具有保护作用,提示中国人群可能为易感人群。