新型冠状病毒中和抗体生物学活性的测定

目的 开发针对新型冠状病毒中和抗体生物学活性及免疫学特性分析的方法。方法 使用生物膜层干涉法(biolayer interferometry, BLI)分析9MW3311中和抗体Fab和S1-RBD的亲和力常数;分别使用酶联免疫吸附法(enzyme linked immunosorbent assay, ELISA)和流式细胞分选法评价中和抗体结合片段(antibody binding fragment, Fab端)和S蛋白的相对结合活性和对ACE2的阻断结合活性;使用假病毒体系评价中和抗体的体外细胞学活性;使用表面等离子共振法(surface plasmon resonance, SPR)测定抗体Fc段与Fcγ和FcRn受体的亲和力常数;使用ELISA法测定抗体Fc段和C1q受体的结合活性;采用PBMC法确定中和抗体的体依赖细胞介导的细胞毒性作用(antibody-dependent cell-mediated cytotoxicity, ADCC)、补体(First subcomponent of the C1 complex)介导的细胞毒性作用(complement depend...     

龙眼壳总黄酮对大鼠局灶性脑缺血再灌注损伤的保护作用及其机制

目的:观察龙眼壳总黄酮对局灶性脑缺血再灌注损伤大鼠的保护作用。方法:120只雄性W istar大鼠随机分为脑缺血再灌注组,龙眼壳总黄酮高、中、低剂量组,尼莫地平对照组,假手术组。给药组术前连续灌胃14d,于末次给药60m in后行手术,采用大脑中动脉局灶性缺血再灌注模型,观察缺血2h再灌注24h后,龙眼壳总黄酮对各药物组大鼠脑梗死体积、脑含水量及脑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)、一氧化氮合酶(NOS)的影响。结果:与脑缺血再灌注模型组相比,龙眼壳总黄酮能够降低大鼠脑梗死体积和脑含水量,抑制脑组织中MDA的生成、降低NO含量、抑制NOS活性、提高SOD的活性。结论:龙眼壳总黄酮对大鼠急性脑缺血再灌注损伤具有保护作用,其机制可能与其抗自由基和抑制NO生成有关。     

关节镜下早期重建与晚期重建前交叉韧带的近期临床疗效比较

目的 比较膝关节前交叉韧带(ACL)损伤后,采用关节镜下自体腘绳肌腱早期重建与晚期重建ACL的临床疗效.方法 回顾性分析2005年7月至2008年7月间采用自体腘绳肌腱重建ACL的95例ACL损伤患者资料,其中早期重建组(伤后2周内实施手术)31例,男21例,女10例;平均年龄为29.3岁.晚期重建组(伤后2～6个月实施手术)64例,男45例,女19例;平均年龄为31.5岁.两组患者的性别、年龄、术前Lysholm评分、国际膝关节文献委员会(IKDC)评定结果等术前指标差异无统计学意义(P>0.05),具有可比性.术后对患者进行体格检查,比较两组患者术后的疼痛消除时间、肿胀消除时间、恢复正常关节活动度(与健侧相同)的时间、Lysholm评分、IKDC评定结果等.结果 早期重建组31例患者术后获平均22.4个月(8～32个月)随访.晚期重建组64例患者术后获平均21.5个月(11～33个月)随访.早期重建组疼痛消除时间[(35.6±3.2)d]、肿胀消除时间[(14.7±2.7)d]、恢复正常关节活动度时间[(48.8±4.8)d]均长于晚期重建组[(21.5±2.6)、(8.5±2.1)、(34.5±4.1)d],差异均有统计学意义(P<0.05).末次随访时膝关节IKDC评定结果:早期重建组A级12例,B级15例,C级4例;晚期重建组A级26例,B级29例,C级9例,两组比较差异无统计学意义(x~2=4.112,P=0.250).两组患者术后末次随访时Lysholm评分平均分别为(89.6±6.9)、(91.5±7.4)分,差异无统计学意义(t=0.677,P=0.500).结论 早期重建与晚期重建ACL的近期临床疗效相近,但早期重建ACL者术后康复时间长于晚期重建者,远期疗效有待于进一步观察.     

Arsenic trioxide Up-Regulate Fas Expression in Human Osteosarcoma Cells

Background Osteosarcoma is a common primary malignant tumor of bone with a poor prognosis due to its propensity for metastasis. The prognosis of patients is highly dependent on the presence or absence of lung metastasis and on the effectiveness of treatment against it. It has been reported that low level expression of Fas protein in human osteosarcoma cell is closely associated with lung metastasis. A large number of studies have shown that arsenic trioxide (ATO) can inhibit proliferation and induce apoptosis of many cancer cell lines; however, its effects on human osteosarcoma cells (Saos-2 cell line) remains unknown. The aim of this study was to investigate the effects of ATO on Saos-2 cells and to characterize its mechanism of Fas-expressing. Methods A group of Saos-2 cells was treated with or without 0.5,1, 2, 4 and 8 μmol/L ATO for three successive days, and the cytotoxicity of ATO was determined by an MTT assay. Morphological changes in cells were studied by Acridine orange / Ethidium bromide (AO/EB ) double staining. Flow cytometry (FCM) was used to assay cell DNA distribution. Another group of cells was pretreated with 10nmol/L Matrix Metalloproteinase7(MMP-7) for 3h. They were then incubated with or without 2 μmol/L ATO for 24h, 48h and 72h. Cytotoxicity, Fas protein and mRNA (add pattern) in levels were systematically studied using MTT, western blot and Real-time PCR, respectively. Cell proliferation, cell cycle progression and apoptosis were examined in this study. Results Proliferation of Saos2 cells was inhibited by ATO in both a dose- and time-dependent manner. The IC50 values at 24, 48 and 72 hours were 9.30, 5.54 and 3.49 μmol/L, respectively. The survival rate of Saos-2 cells in the MMP-7 and ATO co-treated group was significantly higher than the ATO group, but it was lower than the control group. Arsenic trioxide induced G1 phase arrest of the cell cycle and very efficiently stimulated apoptosis in Saos-2 cells, as evidenced by flow cytometric detection of sub-G1 DNA content and AO/EB staining.Western blot results indicated that Fas(FasL) protein expression in osteosarcoma cultures markedly increases in a time dependent manner after exposure to ATO. Compared with control, treatment with ATO 2 μmol/L and 4 μmol/L for 48h , resulted in increase of Fas gene expression to 28.31% and 56.74%, respectively. Our results indicated that ATO induced-apoptosis of Saos-2 cells is mediated through the Fas pathway. Conclusions ATO suppressed cell proliferation of Saos-2 cell in a dose and time-dependent manner and increased Fas protein expression. However, Fas-mediated apoptosis was incompletely interrupted by MMP-7, which suggested that other molecular mechanisms mediate this process. ATO may be used to reduce the risk of metastasis osteosarcomas.     

荧光实时定量PCR定量水体中日本血吸虫尾蚴方法的建立

摘要：目的 建立快速、高效、特异的定量水体中尾蚴的数量和检测水体中日本血吸虫尾蚴残余基因组DNA的方法,来评估水体受日本血吸虫尾蚴污染的程度。方法 根据日本血吸虫基因组DNA中的三个多拷贝序列Sjrh1.0（序列号：U92488.1）、18S小亚基单位核糖体核酸基因(18SrRNA)序列（序列号：AY157226.1）和逆转录转座子SjR2的G55A序列（G55A）（序列号：AF412221.1）,设计常规PCR引物和荧光PCR引物,比较选取较好的靶序列建立SYBR GreenI 实时定量PCR方法,绘制尾蚴数的对数与Ct值的标准曲线,并对疫水中日本血吸虫尾蚴残余的基因组DNA进行检测。结果 尾蚴数的对数与Ct值的标准曲线有良好的线性关系,相关系数r2为0.9186,和重复实验的变异系数小于3%。结论 本方法特异性高,灵敏,可定量水体中尾蚴数,重复性好,对疫水检测有一定的预警作用。     

Fe3O4-As2O3-PLA复合体联合热疗对骨肉瘤治疗的细胞学研究

目的 研究磁性三氧化砷-聚乳酸纳米粒子(Fe₃O₄-As₂O₃-PLA)复合体联合磁流体热疗对骨肉瘤细胞株UMR-106的生长的影响。方法 磁性三氧化砷-聚乳酸纳米粒子复合体的制备,透射电镜及荧光光度计对其形态及载药量进行观察,高频交变磁场中对其进行体外升温实验,骨肉瘤细胞的培养,四甲基偶氮唑蓝(MTT)比色法检测磁性三氧化砷-聚乳酸纳米粒子复合体联合磁流体热疗对骨肉瘤细胞株UMR-106生长的影响,流式细胞仪(FCM)检测凋亡细胞。结果 所制备的磁性三氧化砷-聚乳酸纳米粒子复合体比较均匀,粒径在60~70 nm,载药量约为10.3%,在输出电流Ⅰ=300 A的高频交变磁场中具有升温能力,且能达到肿瘤治疗的有效温度(41℃~46℃),磁性三氧化砷-聚乳酸纳米粒子复合体联合磁性热疗能抑制骨肉瘤细胞株UMR-106的生长,促进其调亡,且较单纯的As₂O₃液组及单纯的Fe₃O₄磁性纳米粒子热疗组的治疗效果明显。结论 Fe₃O₄-As₂O₃-PLA复合体联合磁性热疗可同时发挥As₂O₃的细胞毒性作用和磁感应加热的联合定向治疗作用,效果优于单纯的As₂O₃液组及单纯的Fe₃O₄磁性纳米粒子热疗组,为临床治疗骨肉瘤提供新的治疗方法。     

红花HPLC指纹图谱研究

目的:利用高效液相色谱建立红花的色谱指纹图谱.方法:运用Agilent 1100 DAD-HPLC高效液相色谱仪,采用Kromasil 100-5 C18(4.6mm×250mm,5μm)色谱柱,乙腈-0.1％磷酸梯度洗脱,流速1.0mL/min,检测波长270nm,柱温25℃,进样量10μL,对10批红花进行试验.结果:10批红花中获得17个色谱共有峰,不同产地红花药材指纹图谱相似度有一定差别.结论:采用HPLC方法建立的红花药材指纹图谱,方法稳定、可靠、简便,色谱图展现的各峰分离度较好,特征明显,可作红花药材真伪鉴别标准,但不同种质红花之间存在一定量及质的差异,说明固定种质来源对保障红花质量的稳定性较为重要.     

门急诊军人诊疗流程分析

军队医院着眼保障官兵身心健康,确保为兵服务,确保部队战斗力,是使命所系、职责所在;提高军队医院为军服务水平、规范简化军人伤病员就医诊疗流程和环节,是基本的工作平台。本院自2008年落实《军队医院为兵服务细则》精神以来,不断摸索、     

血小板浓缩物在骨关节病和运动损伤疾病中的应用

Platelet concentrate ( PC ) for surgical use is an evolution of the fibrin glue technologies used since many years. The initial concept of these preparations was to concentrate platelets and their growth factors and activate it into a fibrin gel on a surgical site, in order to improve local healing. The platelets contain alpha granules that are rich in several growth factors, such as platelet-derived growth factor, vascular endothelial growth factor, transforming growth factor-β, insulin-like growth factor, and epidermal growth factor, which play the essential roles in tissue healing and regeneration mechanisms. The relative ease of obtaining and preparation, convenience and applicability in the clinical setting, favorable safety profile and possible beneficial outcomes make PCs a new promising therapeutic approach for future regenerative therapy strategy. Numerous techniques of PC have been developed. The most common forms of autologous platelet concentrates: Platelet rich plasma ( PRP ) and platelet rich fibrin ( PRF ) have been introduced into various medical fields, including orthopedic surgery, plastic surgery, sports injury and other areas of regenerative medicine. More recently, developments in research of the regenerative effects of PC in a range of tissue types including bone, cartilage, tendon and muscle, as well as in the tissues of traumatic injury, have attracted interest in fields such as orthopedic and sport medicine. Many clinical studies suggested that the PC therapies may have benefits to osteoarthropathy such as osteoarthritis and necrosis of femoral head, and the conditions such as epicondylitis, ligament and muscle strains, and tears of rotator cuff, anterior cruciate ligament and Achilles tendon, as well as other sport related injuries. However, our understanding of the mechanisms of PCs in tissue repair and healing is still limited in basic research. There are also limited publications that provided convincing evidences of the efficacy of PCs therapy in the treatment of these injuries/disorders in a randomized controlled clinical trial based study. Therefore, in this review, we describe the various factors that proposed to contribute to the biological activity of PCs, and the published pre-clinical and clinical evidences to support the factors. We also review the current developments on the basic research and clinical application of the PC products in osteoarthropathy and sport medicine, and highlight the areas of emerging clinical applications of PCs.     

多菌型麻风在联合化疗期间发生神经损害及治疗效果的观察

研究MB麻风在MDT期间发生神经损害的情况及评估泼尼松治疗效果。定期对患者的神经功能进行检测并详细填写《麻风患者畸残记录表》;对麻风反应所致的神经损伤亦详细记录;对泼尼松的治疗效果进行全面评估。每例患者在初治前均有多条神经受累,MDT后神经粗大的例数和条数显著减少(P<0.01)。泼尼松对疼痛性神经炎及无疼痛性神经炎均有良好疗效,但对已发生运动功能障碍者效果有限。早期发现和早期治疗麻风病,以及在MDT期间注重对麻风反应的合理有效处置是防治神经功能发生障碍的主要措施和环节。泼尼松对各类神经炎具有良好疗效,但尚需采取其它综合措施以期取得更好效果。