PSMA-CAR-CIK转基因细胞体外杀伤三种前列腺癌细胞株效率的研究北大核心

目的:本研究通过检测前列腺特异性膜抗原(prostate specific membrane antigen,PSMA)-肿瘤特异性嵌合抗原受体(chimeric antigen receptor,CAR)-细胞因子诱导的杀伤细胞(cytokine induced killer cell,CIK)转基因细胞杀伤三种前列腺癌细胞株的效率,鉴定其用于后期治疗前列腺癌的可行性。方法:通过已经构建的二代PSMA-CAR慢病毒表达载体转染CIK细胞,构建PSMA-CAR-CIK转基因细胞;通过CCK8法检测CIK细胞及PSMA-CAR-CIK转基因细胞杀伤三种前列腺癌细胞株PC3、LNCaP、DU145的效率及效靶比。结果:转基因PSMA-CAR-CIK细胞构建成功;3 h~3.5 h为检测效应细胞对靶细胞细胞毒活性时与CCK8试剂孵育的最佳时间;两种效应细胞均在效靶比为10∶1、15∶1、20∶1时,杀伤率逐步增高,无统计学意义(P>0.05),但与其它组相比差异显著(P<0.05)。结论:两种效应细胞对于前列腺癌细胞的杀伤,只要效靶比达到10∶1的比例,就能起到一个较好的杀瘤效果,并且随着效应细胞的增加,其抗肿瘤能力逐步加强;转基因PSMA-CAR-CIK细胞的构建为CAR技术治疗前列腺癌的临床应用奠定了基础。     

基于倾向性评分匹配的聚乙二醇化重组人粒细胞刺激因子预防宫颈癌同步放化疗中性粒细胞减少症效果分析

目的 探讨聚乙二醇化重组人粒细胞刺激因子（PEG-rhG-CSF）在宫颈癌同步放化疗中预防中性粒细胞减少的疗效及安全性。方法 按单中心单臂历史对照研究,选取2019年6月—2021年1月32例符合入排条件,拟行同步放化疗的宫颈癌患者组成试验组,同时搜集2017年1月—2019年6月行同步放化疗且符合对照组治疗方案的宫颈癌患者病历资料,按1∶1倾向性评分匹配法组成对照组。试验组为前瞻性入组,患者第1次化疗给药结束24 h后sc聚乙二醇化重组人粒细胞刺激因子注射液,每次6 mg,每化疗周期1次,共给药2次。若使用PEG-rhG-CSF预防后,中性粒细胞计数（ANC）仍＜1.0×109·L^-1,可给予短效重组人粒细胞刺激因子注射液5 μg·kg^-1,sc给药,直至ANC≥2.0×109·L^-1。对照组患者采用倾向性评分匹配既往同步放化疗的宫颈癌患者,初始不给予PEG-rhG-CSF,当患者出现ANC＜1.0×109·L^-1后sc给予重组人粒细胞刺激因子注射液,5 μg·kg^-1,持续使用,每日1次,直至ANC≥2.0×109·L^-1。记录两组患者3、4度中性粒细胞减少症发生率及持续时间;粒细胞减少性发热（FN）发生率;中性粒细胞减少导致化疗延迟和放疗中断发生率;评价两组不良反应情况,包括乏力、骨关节痛、发热、皮肤黏膜反应、恶心及呕吐等。结果 试验组与对照组各32例,两组基线均衡。试验组中性粒细胞减少症发生率59.4%,对照组为84.3%,试验组显著低于对照组（P＜0.05）;3、4度中性粒细胞减少症发生率试验组为31.3%（10/32）,对照组为56.3%（18/32）,试验组显著低于对照组（P＜0.05）。3度中性粒细胞减少持续时间,试验组中位时间为0 d（0～29 d）,显著低于对照组2 d（0～38 d）;4度中性粒细胞减少持续时间,试验组中位时间为0 d（0～22 d）,显著低于对照组1 d（0～38 d）。两组FN发生率比较无明显差异,但试验组有降低趋势。试验组放疗中断1例,对照组2例,两组比较无显著统计学差异（P＞0.05）;试验组延迟化疗出现4例（12.5%）,对照组出现12例（37.5%）,两组比较差异显著（P＜0.05）。两组不良反应主要包括乏力、骨关节痛、发热、皮肤黏膜反应、恶心及呕吐等,组间比较,差异不显著（P＞0.05）。结论 宫颈癌同步放化疗过程中预防性应用PEG-rhG-CSF具有安全性、有效性,可以降低中性粒细胞减少症发生率,减少FN和不良反应,避免治疗延迟。     

lncRNA-MALAT1/miRNA-145在舒芬太尼预处理减轻大鼠心肌缺血-再灌注损伤中的作用

目的探讨长链非编码RNA(lncRNA)肺腺癌转录子1(MALAT1)在舒芬太尼预处理减轻大鼠心肌缺血-再灌注损伤(MIRI)中的作用。方法 SPF级大鼠36只,体重180~200 g,随机分为三组:假手术组(Sham组),缺血-再灌注损伤组(IR组),缺血-再灌注损伤+舒芬太尼组(SUF组),每组12只。IR组和SUF组建立大鼠心肌缺血-再灌注损伤模型,SUF组于再灌注前10 min尾静脉注射舒芬太尼1μg/kg,Sham组和IR组注射等体积生理盐水。再灌注后2 h检测大鼠血清中肌酸激酶同工酶MB(CK-MB)、心肌肌钙蛋白T(cTnT)和乳酸脱氢酶(LDH)活性,同时检测大鼠心肌梗死面积;检测心肌细胞凋亡相关蛋白Bcl-2、Bax、Cleaved-caspase-3和Caspase-3含量;检测大鼠心肌组织中lncRNA-MALAT1和miRNA-145的相对表达量。结果与Sham组比较,IR组和SUF组CK-MB和cTnT浓度明显升高,LDH活性明显增强,心肌梗死面积明显增大,心肌组织中Bax/Bcl-2比值明显降低,Cleaved-caspase-3蛋白含量明显升高,lncRNA-MALAT1相对表达量明显升高,而miRNA-145相对表达量明显降低(P<0.05)。与IR组比较,SUF组CK-MB和cTnT浓度明显降低,LDH活性明显减弱,心肌梗死面积明显减小,Bax/Bcl-2比值明显升高,Cleaved-caspase-3蛋白含量明显降低,lncRNA-MALAT1相对表达量明显降低,miRNA-145相对表达量明显增加(P<0.05)。结论舒芬太尼预处理能减轻大鼠心肌缺血-再灌注损伤程度,其机制可能与抑制lncRNA-MALAT1、升高miRNA-145表达有关。     

眼轮匝肌下脂肪垫的解剖学及整形外科学的研究

目的：研究眼轮匝肌下脂肪垫的解剖结构,和相邻组织结构的关系,评价其在眼部年轻化手术中的应用效果。方法：对成人头颅部人体标本7具（14侧眼部标本）进行眶部解剖研究;对50例术前有明显面部老化征象患者进行了眼部手术,手术中将眼轮匝肌下脂肪垫部分去除。结果：在眼轮匝肌下和上颌骨、颧骨骨膜的浅面,存在有脂肪垫,其大部分分布在眼眶外围,并与周围脂肪垫相连续。50例眼袋患者,术后下睑区平展,眶隔脂肪袋消失,下眶缘沟消失,眶下外侧骨缘轮廓平和。其中19例伴有上睑臃肿下垂患者,术后眼裂增宽,眼睑变薄,无上睑外侧臃肿表现。结论：眼部年轻化手术适当去除眼轮匝肌下脂肪垫,可明显改善手术效果。     

老年人工股骨头置换术后综合抗骨质疏松治疗的近期疗效分析

目的探讨老年人工股骨头置换患者术后抗骨质疏松治疗方法,以提高其治疗水平。方法对14例人工股骨头置换术术后患者进行包括健康宣教、生活习惯调整、抗骨质疏松药物、物理治疗、康复运动治疗及家庭康复指导等的综合治疗,并测量患者术前、术后6个月骨密度及Harris评分。结果 14例人工股骨头置换患者治疗效果良好,骨密度由术前平均0.64±0.05g/cm2增加至0.65±0.05g/cm2,术前、术后骨密度变化无显著性差异(t=1.51,P=0.16),但有效阻止了骨折患者创伤后的骨量丢失;Harris评分由术前平均48.93±4.63分增加至90.79±2.94分,术前、术后评分具有显著性差异(t=37.75,P<0.05),Harris评分略高于国内同类研究。结论综合抗骨质疏松治疗有利于提高老年人工股骨头置换患者骨密度和生活质量。     

Role of GSTP1 Ile105Val and XRCC1 Arg194Trp,Arg280His and Arg399Gln gene polymorphisms in the clinical outcome of advanced non-small cell lung cancer

We conducted a study to investigate the association between the clinical outcome and GSTP1 Ile105Val and XRCC1 Arg194Trp, Arg280His and Arg399Gln gene polymorphisms in advanced NSCLC patients with cisplatin-based chemotherapy. Between January 2010 and December 2012, a total of 206 patients with advanced NSCLC were histopathologically confirmed were included into analysis. By logistic regression analysis, individuals carrying the AG and GG genotypes of GSTP1 Ile105Val were associated with better response to chemotherapy when compared with the AA genotype, and the adjusted Ors (95% CI) were 2.06 (1.10-3.86) and 4.89 (1.52-18.33), respectively. The TT genotype of XRCC1 Arg194Trp was correlated with better response to chemotherapy compared to the CC genotype, and the adjusted OR (95% CI) was 3.23 (1.20-9.30). By Cox Hazard Proportional Model, the GG genotype of GSTP1 Ile105Val and the TT genotype of XRCC1 Arg194Trp were found to be associated with lower risk of death from all causes when compared with the wide-type genotype, and the adjusted HRs (95% CI) were 0.05 (0.01-0.18) and 0.20 (0.07-0.62), respectively. Moreover, individuals carrying both the G/A+G/G genotype of GSTP1 Ile105Val and the G/A+A/A of XRCC1 Arg194Trp were associated with heavy greater CR+PR response to chemotherapy (OR=2.98, 95% CI=1.39-6.42), and also correlated with longer overall survival of advanced NSCLC (HR=0.19, 95% CI=0.05-0.61). In conclusion, we found that the GSTP1 Ile105Val and XRCC1 Arg194Trp were associated with better response to chemotherapy and longer survival of advanced NSCLC, compared to the wide-type genotype.     

金芪降糖片组分体外抗糖尿病的作用及机制

目的:探讨金芪降糖片各组分体外抗糖尿病的作用及机制。方法:采用DPPH自由基清除,活性氧自由基清除,AGEs形成抑制,α-糖苷酶抑制,醛糖还原酶抑制,脂肪酶抑制,脂多糖(LPS)诱发小鼠腹腔巨噬细胞活化一氧化氮(NO)释放抑制建立系统的体外抗糖尿病活性评价筛选模型,考察金芪降糖片的4个标准组分芪总黄酮、黄芪总皂苷、金银花总有机酸、黄连总生物碱对以上各模型的作用。结果:黄芪总黄酮、金银花总有机酸、黄连总生物碱具有清除DPPH自由基和活性氧自由基活性;各组分对AGEs形成无明显抑制活性;黄芪总黄酮、金银花总有机酸对糖苷酶有抑制活性;黄芪总黄酮、黄芪总皂苷、金银花总有机酸对醛糖还原酶有抑制活性;黄芪总黄酮、黄芪总皂苷、金银花总有机酸对胰脂肪酶具有抑制活性;黄芪总皂苷、黄连总生物碱对LPS诱导的小鼠腹腔巨噬细胞释放NO有抑制作用。结论:金芪降糖片具有多组分多靶点协同抗糖尿病的作用,机制涉及抑制α-糖苷酶、抑制脂肪酶、清除自由基、抑制醛糖还原酶、抑制NO释放。     

Nano-hydroxyapatite modulates osteoblast lineage commitment by stimulation of DNA methylation and regulation of gene expression

Hydroxyapatite (HA) is the primary structural component of the skeleton and dentition. Under biological conditions, HA does not occur spontaneously and therefore must be actively synthesized by mineralizing cells such as osteoblasts. The mechanism(s) by which HA is actively synthesized by cells and deposited to create a mineralized matrix are not fully understood and the consequences of mineralization on cell function are even less well understood. HA can be chemically synthesized (HAp) and is therefore currently being investigated as a promising therapeutic biomaterial for use as a functional scaffold and implant coating for skeletal repair and dental applications. Here we investigated the biological effects of nano-HAp (10 × 100 nm) on the lineage commitment and differentiation of bone forming osteoblasts. Exposure of early stage differentiating osteoblasts resulted in dramatic and sustained changes in gene expression, both increased and decreased, whereas later stage osteoblasts were much less responsive. Analysis of the promoter region one of the most responsive genes, alkaline phosphatase, identified the stimulation of DNA methylation following cell exposure to nano-HAp. Collectively, the results reveal the novel epigenetic regulation of cell function by nano-HAp which has significant implication on lineage determination as well as identifying a novel potential therapeutic use of nanomaterials.