DNA flow-cytometric analysis in colorectal cancer: A comparison of metastasizing and non-metastasizing tumours

The most common cause of death in patients with colorectal cancer is metastatic liver disease. In order to identify patients at a high risk of developing hepatic secondaries from colorectal cancers, DNA content was measured in metastasizing colorectal primaries (Group I, n= 32) as well as in their subsequently resected liver secondaries and in sections of non-metastasizing colorectal cancers (Group II, n= 25). A modified interpretation system involving both a DNA index and percentage of cycling cells (those in S and G2 + M phases) was developed. DNA content was measured in paraffin-embedded sections by flow cytometry using internal controls (human peripheral blood mononuclear cells) and non-malignant tissue controls (19 patients with diverticular disease). In Group I there were significantly more tumours with both abnormal ploidy (aneuploid or abnormal tetraploid peak) and > 15% cycling cells compared with Group II (Chi-squared; P= 0.034). The combination of abnormal ploidy and > 15% cycling cells was superior to Dukes’ classification for identifying metastasizing tumours (Logistic Regression; P= 0.047). However, it was not possible to discriminate between the two groups using either DNA ploidy or the percentage of cycling cells alone. The metastasizing colorectal cancers exhibited similar DNA ploidy characteristics and had a similar percentage of cycling cells compared with their liver metastases. These results suggest that tumour DNA ploidy plus the percentage of cycling cells may predict the development of liver metastases and thus survival in patients with colorectal cancer.     

Cloning of Human α-defensin-1 (HNP-1) Gene and Construction of Its Eukaryotic Expression Vector

1 Introduction Defensins are small cationic antimicrobial peptides that function in the host's innate immune system. The human defensin family includes three small peptides from the azurophil granules of polymorphonuclear cells named human neutrophil peptide (HNP)-1, HNP-2, HNP-3,which consist 5%-7% of the protein of human neutrophil. HNP-4 is approximately one hundred times less abundant. They demonstrate antibacterial, antifungal and antiviral properties in vitro. HNPs are important component of nonoxidative mechanism in macrophages, and can direct inactivate the enveloped viruses. Because only special cells express defensins. And it is hard to extract them naturally and the production is few. So researcher expect to obtain defensins highly through heterogenous expression by gene engineering technology. In order to express HNP-1, we cloned the cDNA of HNP-1 from human polymorphonuclear cells in peripheral blood, and constructed its eukaryotic expression vector, which provided a base for the further study on its mechanism of antimicrobial effect.     

白细胞介素-4和γ-干扰素基因联合治疗胶质瘤

目的 探讨逆转录病毒载体介导的白细胞介素(IL)-4和γ-干扰素(IFN)基因联合治疗胶质瘤的作用。方法构建携带IL-4或IFN-γ基因的逆转录病毒载体，将其导入逆转录病毒包装细胞；将携带IL-4和IFN-γ基因的逆转录病毒包装细胞分别或联合注射到脑内荷瘤大鼠的肿瘤组织中，观察其治疗作用。结果成功获得携带治疗基因的逆转录病毒包装细胞PA317IFN-γ和PA317IL-4，所产病毒滴度分别为2x10^6cfu／ml和2．5x10^6cfu／ml。包装细胞瘤内注射能够诱导大鼠产生抗肿瘤免疫反应，杀伤肿瘤细胞，联合应用具有协同治疗作用。结论携带IL-4或IFN-γ基因的逆转录病毒包装细胞瘤内注射是一种有效的胶质瘤基因治疗方法，两者联合应用具有协同治疗作用。     

稳定增殖神经干细胞的实验研究

目的探讨稳定、可靠建立神经干细胞体外增殖的方法,并对增殖培养的细胞进行鉴定。方法获取胚胎大鼠的脑组织,通过加入神经生长因子和采用保留细胞联系技术操作,使脑组织中的神经干细胞在体外克隆增殖并稳定传代。以免疫荧光方法对增殖克隆的神经干细胞球进行鉴定。结果神经干细胞不断增殖形成神经干细胞球且神经干细胞能快速稳定传代增殖。培养的细胞为神经干细胞特异性巢蛋白(nestin)染色阳性细胞。结论在神经生长因子的作用下,利用保留细胞联系技术操作,神经干细胞可以在体外快速、稳定克隆增殖并传代。     

国产鼻喷鲑鱼降钙素治疗骨质疏松性疼痛的疗效观察

目的观察国产鼻喷鲑鱼降钙素治疗骨质疏松性疼痛的疗效。方法采用随机分组对照设计。选择骨质疏松患者53例,随机分为两组,治疗组(30例)给予鼻喷鲑鱼降钙素,20u/次,1次/天,8周为1个疗程,停药后继续观察3周。对照组(23例)口服钙尔奇D片,1片/次,1次/天,共8周;采用国际惯用分级标准对疼痛予以评估。结果治疗组患者的疼痛明显改善,其总有效率为95%;在停药后缓解疼痛的作用仍可维持4周以上,Ⅲ度骨质疏松患者的治疗效果较Ⅰ度和Ⅱ度患者的效果差;对照组症状未见明显改善。结论用国产鼻喷鲑鱼降钙素治疗骨质疏松性骨痛,可明显缓解患者的疼痛症状。     

猪尾导管连接化疗药盒实现肝脏肿瘤多重化疗的可行性研究

目的 探讨使用猪尾造影导管与单一化疗药盒连接实现肝脏肿瘤多重化疗的可行性及初步疗效。资料与方法 对30例晚期肝脏恶性肿瘤患者采用经股动脉穿刺、腹腔动脉内植入猪尾导管与皮下埋置药盒并序贯化疗，统计手术成功率、并发症及临床疗效。结果 30例手术全部成功，发生切口延迟愈合1例。术后随访28例，经影像学复查对比，病变缩小7例，稳定12例，进展9例。随访期间12例患者死亡，术后生存期42d-10个月，中位生存期6个月。另外16例患者至今存活，已达术后2-16个月。结论 采用猪尾造影导管置于腹腔动脉并与化疗药盒相连接，能够对晚期肝脏恶性肿瘤实现多重化疗，技术上简单可行，并发症少，初步疗效令人满意。     

急性脑出血患者血清白介素-6和转化生长因子-β1含量的变化及其临床意义

目的探讨急性脑出血患者血清白介素-6(IL-6)和转化生长因子β1(TGF-β1)水平在发病过程中的动态变化及其临床意义.方法采用双抗体夹心法检测30名正常人(对照组)和30例急性脑出血患者病后第1、3、7、14 d的血清IL-6和TGF-β1浓度.结果脑出血组在病程第1、3、7 d血清IL-6水平均比对照组明显增高(均P<0.05),TGF-β1水平均明显减低(均P<0.05),至第14 d血清IL-6、TGF-β1水平均接近正常水平.血清IL-6水平在病程第1 d达到峰值(P<0.05),TGF-β1在病程第3 d为低谷值(P<0.05).大量脑出血患者血清IL-6水平均明显高于中、小量出血患者(均P<0.05).不同出血量患者之间TGF-β1水平的差异无显著性(P>0.05).神经功能缺损程度与血清IL-6水平呈正相关(r＝0.625,P<0.05),与TGF-β1水平无关(r＝0.001,P>0.05).IL-6水平与TGF-β1水平无明显相关性.结论 IL-6、TGF-β1与脑出血的病理过程有密切关系.血清IL-6浓度测定对判断脑出血的病情严重程度有一定意义.     

Brevibacterium sp.DGCDC-82发酵罐中生产胆固醇氧化酶

用Brevibacterium sp.DGCDC-82在7L发酵罐中分批培养.分别对添加剂吐温-80、培养温度、培养基的pH、通风量和搅拌速度在胆固醇氧化酶的生产中的影响进行了研究.结果显示以上条件均对产酶有影响.在不同的发酵阶段改变发酵操作条件,发酵20 h最高酶活可达1203U/L,生产强度可达60 U/(L·h).既有效地提高了胆固醇氧化酶的产量,又防止了发酵过程中的泡沫外溢.     

假单胞菌胆固醇酯酶的生产Ⅰ.菌种筛选和发酵条件的研究

通过平板初筛和摇瓶复筛,从371株菌中筛得一株有工业生产价值的胆固醇酯酶产生菌,并对它进行了发酵条件的研究。用所获得的胆固醇酯酶配制的三酶试剂具有良好的稳定性,适用于临床检测。     

成牙本质细胞中上游刺激因子1对骨桥素表达调控的研究

目的检测成牙本质细胞中上游刺激因子1（upstream stimulatory factor 1，USF1）对骨桥素表达的影响。方法培养成牙本质细胞MDPC-23，稳定转染PCMV-USF1和酸性-USF（A-USF）质粒，提取总RNA，半定量反转录聚合酶链反应检测骨桥素的表达水平，并对结果进行统计学分析。结果筛选出稳定转染USF1和A-USF的抗性克隆，USF1、A-USF转染组和对照组骨桥素相对灰度比值分别为：60．33％±4．51％、229．33％±7．09％、110．00％±15．62％，转染组与对照组差异有统计学意义（P〈0．01）。结论USF1可调控成牙本质细胞骨桥素mRNA转录，该作用被A-USF部分阻断。