杜仲细胞悬浮培养产黄酮及其动力学研究

本文应用正交设计对杜仲细胞悬浮培养的基本培养基和植物生长物质浓度进行了筛选,并对影响杜仲细胞悬浮培养和总黄酮含量的不同因素进行了考察。结果表明,B5培养基+0.5mg/L NAA+0.6mg/L 6-BA、蔗糖30g/L、初始pH 5.0-5.5、接种量20g(FW)/L以及摇床转速110r/min为杜仲细胞悬浮培养的适宜条件。通过对杜仲悬浮细胞生长和代谢动力学的分析表明：杜仲细胞悬浮培养生长符合Logistic生长模型,最大比生长速率( m)为0.417d-1;细胞基于蔗糖的真正比生长得率(YG)与维持系数(m)分别为0.619g/g和0.0206g/(g·d-1);黄酮合成属部分生长耦联型,可用Luedeking-Piret模型进行描述。研究结果为杜仲细胞大规模悬浮培养生产天然活性成分奠定了基础。     

Adhesion of lymphoid cell lines to fibronectin-coated substratum: Biochemical and physiological characterization and the identification of a 140-kDa fibronectin receptor

Little information is available on the interaction between lymphocytes and fibronectin (fn). To gain a better understanding on this issue we examined the adhesion of 12 lymphoid cell lines, each exhibiting different phenotypic characteristics, to fn-coated substratum. Of the cell lines tested, five that adhered to fn possessed B-cell characteristics, while neither the T-cell lines nor the pre-B-cell line adhered. The physiology and biochemistry of adhesion of a B-cell line, MOPC 315, were examined in detail. Our results indicated that (1) the adhesion was a specific and time-dependent process, (2) the adhesion was temperature-dependent and inhibited by metabolic inhibitors, such as KCN and 2-deoxyglucose, (3) the presence of cycloheximide and pretreatment of cells with trypsin inhibited adhesion, (4) a 140-kDa surface protein was immunoprecipitated by anti-fn receptor antibodies, (5) the presence of divalent cations was essential for adhesion, (6) the presence of colchicine had no effect on adhesion, while cytochalasin B partially inhibited adhesion, and (7) the treatment of cells by both phorbol 12-myristate 13-acetate and calcium ionophore A23187 enhanced adhesion. In this study, we have established the interaction between lymphoid cell lines and fn. Such an interaction might play an important role in the behavior of lymphocytes in tissues.     

The discovery of anthranilic acid-based MMP inhibitors. Part 3: incorporation of basic amines.

Anthranilic acid derivatives bearing basic amines were prepared and evaluated in vitro and in vivo as inhibitors of MMP-1, MMP-9, MMP-13, and TACE. Piperazine 4u has been identified as a potent, selective, orally active inhibitor of MMP-9 and MMP-13.     

Clonality analysis of archival cervical smears. Correlation of monoclonality with grade and clinical behavior of cervical intraepithelial neoplasia

OBJECTIVE: To establish a polymerase chain reaction (PCR)-based clonality assay for archival cervical smears and examine its value in the detection of cervical intraepithelial neoplasia (CIN) and prediction of its clinical behavior. STUDY DESIGN: Dyskaryotic cells were microdissected from archival cervical smears of 33 cases and subjected to PCR-based clonality analysis of the androgen receptor gene. High-risk HPV subtypes were screened by PCR. RESULTS: Monoclonal patterns were found in 9/9 CIN 3 and 15/21 CIN 2, while polyclonal patterns were observed in the remaining 6 CIN 2 and 3/3 CIN 1. All patients with monoclonal CIN lesions, including 15 CIN 2, showed recurrence of the disease despite treatment. The original CIN 2 and recurrent CIN lesion in each of the 6 examined cases showed the same monoclonal pattern, suggesting a clonal link. In contrast, the patients with polyclonal CIN 1 or 2 became negative and remained disease free. High-risk HPV subtypes were found in all monoclonal CIN lesions, including 9 CIN 3 and 15 CIN 2, and in 4/6 polyclonal CIN 2 but not in CIN 1 lesions. CONCLUSION: Clonality analysis of cervical smears is potentially valuable in the identification of true neoplastic cells and prediction of clinical behavior of CIN 2 lesions.     

Enhancement of hippocampal neurogenesis by lithium

Increasing evidence suggests that mood disorders are associated with a reduction in regional CNS volume and neuronal and glial cell atrophy or loss. Lithium, a mainstay in the treatment of mood disorders, has recently been demonstrated to robustly increase the levels of the cytoprotective B-cell lymphoma protein-2 (bcl-2) in areas of rodent brain and in cultured cells. In view of bcl-2's antiapoptotic and neurotrophic effects, the present study was undertaken to determine if lithium affects neurogenesis in the adult rodent hippocampus. Mice were chronically treated with lithium, and 5-bromo-2-deoxyuridine (BrdU) labeling of dividing cells was conducted over 12 days. Immunohistochemical analysis was undertaken 1 day after the last injection, and three-dimensional stereological cell counting revealed that lithium produced a significant 25% increase in the BrdU-labeled cells in the dentate gyrus. Double-labeling immunofluorescence studies were undertaken to co-localize BrdU-positive cells with neuron-specific nuclear protein and showed that approximately 65% of the cells were double-labeled. These results add to the growing body of evidence suggesting that mood stabilizers and antidepressants exert neurotrophic effects and may therefore be of use in the long-term treatment of other neuropsychiatric disorders.     

植物转录抑制子的结构特征及其作用机理

转录因子依转录调控能力可分为激活子和抑制子。植物转录抑制蛋白的分类依据很多,从作用方式上可分为主动抑制子和被动抑制子两大类：根据与DNA结合的方式则可分为锌指类、MYB类、AP2／EREBP类、bHLH类和bZlP类等。植物主动抑制子通过其含有的抑制域对转录直接起抑制作用。抑制域又可分很多类,但多数为含有类似EAR基序的保守性基序,其上具有几个保守性亮氨酸残基。植物转录抑制子主要通过对激活子或基本转录复合物产生作用及改变染色体结构3种方式来抑制目标基因的转录。有关植物转录抑制子的研究虽很欠缺,但以拟南芥SUPERMAN等抑制子的EAR基序为代表的研究表明,抑制域是阐明植物转录抑制子功能和下游基因表达调控机理的核心对象,而融合抑制子沉默技术（CRES-T）也为人为调控基因沉默带来了新的技术手段。     

蛇毒类凝血酶鼠单克隆抗体的制备

采用杂交瘤技术,获得了４株稳定分泌抗蛇毒类凝血酶的单克隆抗体杂交瘤细胞株,均属ＩｇＧ１ｋ链,４株杂交瘤细胞培养上清液效价为 ４ × １０－１～４ × １０－２,腹水效价为 ４ × １０－１～３．２ ×１０－５。