分子激发态寿命及其应用研究

寿命测量现在使用的有位相偏移法和脉冲法。本工作是利用脉冲法进行的。本论文做了三方面的工作：1.分子激发态寿命在分析化学中的应用。荧光寿命是荧光物质的特征参数，可利用来进行定性，定量分析工作。从荧光强度与荧光寿命的基本公式I＝A_ie~(-t/τi)出发，可以得到c_1/c_2=(a_1τ_1ε_2φ_2)/(α_2τ_2ε_1φ_1)其中C_1，C_2是同一体系中两种不同物质的浓度，τ是荧光寿命，ε为激发波长下的消光系数，φ为量子产率，A是幅度常数。ε与φ可预先测量或查表得到。τ，A可根据荧光衰减曲线与标准衰减函数相拟合，解卷积后同时得到。因此可用上式进行定量分析。本论文利用上式分析了卟吩类化合物，其中包括四苯基卟吩；四－邻氯苯基卟吩；锌－四苯基卟吩络合物在不同浓度下的组合，分析误差绝大多数在10％以内。从（1）式出发得到另一个基本公式A＝KCT其中T为时间相关单光子计数时间，K是一常数。根据（3）式进行了多环芳香碳氢化合物分析。稀土发光材料的荧光寿命研究。本工作者首先使用不同的激发光源，不同分光装置和不同的讯号探没器及讯号处理方式，建立了测量范围较宽的荧光寿命测量装置。测量了Eu~(3+), Eu~(2+), Ce~(3+), Dy~(3+), Nd~(3+), Tb~(3+)在不同基质中的荧光寿命。其数值从10~(-8)秒到10~(-3)秒，探测波长包括可见和近红外，并进行了要必的讨论。直接测定单线态氧在溶液中寿命的研究。本工作装置了一套近红外弱信号探测系统测定了O_2(~1△_g)在液相中的磷光寿命。

盐和有机物对氨基酸热力学性质的影响-NaCl氨基酸-葡萄糖-水体系

在278.15-318.15范围内，本文测定了以下四个无液接电池的电动势：Pt,H_2(g, 1 atm)|HCl(m)、X Mass% Glucose-H_2O|Ag-AgCl (A) Pt,H_2 (g, 1 atm)|G(m_1), HGCl (m_2), X Mass% Glucose-H_2O|Ag-AgCl (B) Pt,H_2 (g, 1 atm) |HCl (m), NaCl (M-m), X Mass% Glucose-H_2O|Ag-AgCl (C) Pt,H_2 (g, 1 atm)|G(m_1), HGCl (m_2), NaCl (M-m), X Mass% Glucose-H_2O|Ag-AgCl (D) 其中G为中性甘氨酸，NH_3CH_2COO~-, HGClm为甘氨酸的盐酸盐，为相应电解质的质量摩尔浓度，X为葡萄糖在葡萄糖-水混合溶剂中的质量百分数，M为恒定的离子强度且M = 1.0mol/kg。并且测定了Glucose-H_2O的密度和介电常数。利用传统的D-H公式外推法和基于Pitzer理论的多项式逼近法分别确定了电池的标准电动势E°_3、E°_3，以及甘氨酸的一级热力学解离1.0mol/kg）-葡萄糖-水-HCl的HCl无限稀释溶液为参考态，并将两种方法得到的结果作了比较。甘氨酸的一级热力学解离常数符合Harned-Robinson方程：pK = A_1/T + A_2 + A+3 T 本文讨论了盐和有机物对pK_1的影响，并根据前人及我们的工作，指出在极性质子溶剂中和在极性非质子溶剂中的pK_1对1/D作图，分别得到直线和曲线。同时讨论了混合溶剂中甘氨酸的一级解离过程的各个热力学量ΔG°，ΔH°，ΔS°，ΔCp°，并讨论HCl的迁移性质和有机物葡萄糖、盐对它们的影响。最后将不同混合溶剂中甘氨酸解离过程的迁移能，迁移熵等作了比较。

生长在尖晶石衬底上的GaN外延层的喇曼散射研究

在室温下测量了用MOVPE方法生长在尖晶石(MgAl_2O_4)衬底上的GaN外延层的一阶喇曼光谱。应用各种背散射和90°散射配置，测得了除低频E_2模外所有GaN的喇曼活性光学声子模。并且在X(Z,X)Z和X(Y,Y)Z配置下观测到了由A_1和E_2模混合形成的准TO和准LO模。所得结果与群论选择定则预计的一致。

长白山北坡针叶林下土壤淋洗液的化学性质及土壤特性

为了研究长白山北坡针叶林下土壤淋洗液的化学性质及土壤特性。我们于1987年6-9月，在红松云冷杉林。岳桦云冷杉林标准地，收集了大气降水，林内降水及土壤淋洗液，进行化学分析。结果表明：在红松云冷杉林标准地大气降水经过林冠后产生酸化作用。在红松云冷杉林和岳桦云冷杉林标准地，通过A_0层淋洗液进入土壤A_1层的养分元素（公斤/公顷）为：K 7.4-11.0, Ca 5.7-5.8, Mg 1.1-1.2，有机碳120.4-156.1，全氮1.9-3.4, SO_4-S 1.8-3.6。 A_0+A_1层养分元素的淋失量（公斤/公顷）为：K 1.2-1.6, Ca 2.2-4.6, Mg 0.5-0.9，有机碳35.4-43.9，全氮0.6-0.7, SO_4-S 1.2-2.7。A_0层淋洗液酸度大（pH5.2-5.6）。有机酸含量高（有机碳46-62mg/1）。Fe、Al、Si均从A_1层淋洗下来，并在其下的淀积层（B）淀积。因此，本区针叶林下土壤具有灰化过程的典型特征。土壤分析结果表明：长白山北坡暗针叶带土壤，具有灰化过程，其中杜香落叶松林下土壤具有明显的灰土淀积层，在分类上属灰土。

长白山阔叶红松林土壤氮转化微生物作用的研究

自长白山北坡自然保护区采集阔叶红松林 A_0 和 A_1 层土壤。进行了微生物数量、土壤酶活性、微生物生物量、土壤速效氮及土壤中群体微生物氮转化测定；分离并鉴定了芽孢杆菌36株、产荧光假单胞菌34株，并对各优势菌株进行了氮转化活性的测定。A_0 层土壤在微生物数量、土壤酶活性、微生物生物量、土壤速效氮 (NH_4~+-N) 等方面明显高于A_1层。氨化作用、硝化作用速率也得到同样的结果；硝酸盐还原、固氮作用及同化作用速率两次采样测定的结果不同。而土壤速效氮 (NO_3~－N)是 A_1 高于 A_0层。芽孢杆菌的优势种是 B. megaterium 和B. cereus，产荧光假单胞菌的优势种是 P. fluorescens－F。所有分离到的芽孢杆菌及产荧光假单胞菌均能活跃地进行氨化作用。所有芽孢杆菌都能进行反硝化作用。而能进行反硝化作用的 P. fluorescens－F及 P. fluorescens－C其作用能力均高于 Bacillus S的20倍以上，同化作用是这两类菌的共同特征。不同种之间以及同种异株间进行氮转化速率的比较。有的差异很大。讨论了这两类菌的数量分布与土壤氮转化速率的关系。建立了反映土壤内部氮转化的模型。

Expression, purification and spectral characterization of p21(Waf1/Cip1)

p21(Waf1/Cip1), best known as a broad-specificity inhibitor of cyclin/cyclin-dependent kinase complexes, can interact with various target proteins, and this ability relies on its structural plasticity. Therefore, studies on the structural properties of p(21) are very important to understand its structure-function relationship. However, detailed studies on its secondary structure and biophysical properties have been comparatively sparse. A human p(21) gene was cloned into the temperature expression vector pBV220 and transformed into Escherichia coli strain JM109.

谢聂稳定判据

谢绪恺、聂义勇判据的提出 比国外学者的同类发现要早得多。由于历史原因,这判据过去未曾宣扬。81年本刊上的简要介绍 曾引起了我国广大读者的重视和兴趣。现在,应读者要求,再作一次较详细的介绍。谢聂判据,简单地说,就是:(A)给多项式式 a_0x~n+a_1x~(n-1)+…+a_n 或 a_0+a_1+…+a_nx~n(a_i>0,i=0,1,2,…n)定义一判定系数α_i=a_i-|a|+2/a_ia_i+1,(B)多项式所表征的系统的稳定性由这判定系数α_i 判定,稳定的必要条件是α_(?)

Gene expression disruptions of organism versus organ in Drosophila species hybrids.

Hybrid dysfunctions, such as sterility, may result in part from disruptions in the regulation of gene expression. Studies of hybrids within the Drosophila simulans clade have reported genes expressed above or below the expression observed in their parent species, and such misexpression is associated with male sterility in multigenerational backcross hybrids. However, these studies often examined whole bodies rather than testes or had limited replication using less-sensitive but global techniques. Here, we use a new RNA isolation technique to re-examine hybrid gene expression disruptions in both testes and whole bodies from single Drosophila males by real-time quantitative RT-PCR. We find two early-spermatogenesis transcripts are underexpressed in hybrid whole-bodies but not in assays of testes alone, while two late-spermatogenesis transcripts seem to be underexpressed in both whole-bodies and testes alone. Although the number of transcripts surveyed is limited, these results provide some support for a previous hypothesis that the spermatogenesis pathway in these sterile hybrids may be disrupted sometime after the expression of the early meiotic arrest genes.

A novel, non-apoptotic role for Scythe/BAT3: a functional switch between the pro- and anti-proliferative roles of p21 during the cell cycle.

BACKGROUND: Scythe/BAT3 is a member of the BAG protein family whose role in apoptosis has been extensively studied. However, since the developmental defects observed in Bat3-null mouse embryos cannot be explained solely by defects in apoptosis, we investigated whether BAT3 is also involved in cell-cycle progression. METHODS/PRINCIPAL FINDINGS: Using a stable-inducible Bat3-knockdown cellular system, we demonstrated that reduced BAT3 protein level causes a delay in both G1/S transition and G2/M progression. Concurrent with these changes in cell-cycle progression, we observed a reduction in the turnover and phosphorylation of the CDK inhibitor p21, which is best known as an inhibitor of DNA replication; however, phosphorylated p21 has also been shown to promote G2/M progression. Our findings indicate that in Bat3-knockdown cells, p21 continues to be synthesized during cell-cycle phases that do not normally require p21, resulting in p21 protein accumulation and a subsequent delay in cell-cycle progression. Finally, we showed that BAT3 co-localizes with p21 during the cell cycle and is required for the translocation of p21 from the cytoplasm to the nucleus during the G1/S transition and G2/M progression. CONCLUSION: Our study reveals a novel, non-apoptotic role for BAT3 in cell-cycle regulation. By maintaining a low p21 protein level during the G1/S transition, BAT3 counteracts the inhibitory effect of p21 on DNA replication and thus enables the cells to progress from G1 to S phase. Conversely, during G2/M progression, BAT3 facilitates p21 phosphorylation by cyclin A/Cdk2, an event required for G2/M progression. BAT3 modulates these pro- and anti-proliferative roles of p21 at least in part by regulating cyclin A abundance, as well as p21 translocation between the cytoplasm and the nucleus to ensure that it functions in the appropriate intracellular compartment during each phase of the cell cycle.

Inhibition of the anaphase-promoting complex by the Xnf7 ubiquitin ligase.

Degradation of specific protein substrates by the anaphase-promoting complex/cyclosome (APC) is critical for mitotic exit. We have identified the protein Xenopus nuclear factor 7 (Xnf7) as a novel APC inhibitor able to regulate the timing of exit from mitosis. Immunodepletion of Xnf7 from Xenopus laevis egg extracts accelerated the degradation of APC substrates cyclin B1, cyclin B2, and securin upon release from cytostatic factor arrest, whereas excess Xnf7 inhibited APC activity. Interestingly, Xnf7 exhibited intrinsic ubiquitin ligase activity, and this activity was required for APC inhibition. Unlike other reported APC inhibitors, Xnf7 did not associate with Cdc20, but rather bound directly to core subunits of the APC. Furthermore, Xnf7 was required for spindle assembly checkpoint function in egg extracts. These data suggest that Xnf7 is an APC inhibitor able to link spindle status to the APC through direct association with APC core components.

Ubiquitylation of p53 by the APC/C inhibitor Trim39.

Tripartite motif 39 (Trim39) is a RING domain-containing E3 ubiquitin ligase able to inhibit the anaphase-promoting complex (APC/C) directly. Through analysis of Trim39 function in p53-positive and p53-negative cells, we have found, surprisingly, that p53-positive cells lacking Trim39 could not traverse the G1/S transition. This effect did not result from disinhibition of the APC/C. Moreover, although Trim39 loss inhibited etoposide-induced apoptosis in p53-negative cells, apoptosis was enhanced by Trim39 knockdown in p53-positive cells. Furthermore, we show here that the Trim39 can directly bind and ubiquitylate p53 in vitro and in vivo, leading to p53 degradation. Depletion of Trim39 significantly increased p53 protein levels and cell growth retardation in multiple cell lines. We found that the relative importance of Trim39 and the well-characterized p53-directed E3 ligase, murine double minute 2 (MDM2), varied between cell types. In cells that were relatively insensitive to the MDM2 inhibitor, nutlin-3a, apoptosis could be markedly enhanced by siRNA directed against Trim39. As such, Trim39 may serve as a potential therapeutic target in tumors with WT p53 when MDM2 inhibition is insufficient to elevate p53 levels and apoptosis.

IGF-1 or insulin, and the TSH cyclic AMP cascade separately control dog and human thyroid cell growth and DNA synthesis, and complement each other in inducing mitogenesis.

The regular doubling of cell mass, and therefore of cell protein content, is required for repetitive cell divisions. Preliminary observations have shown that in dog thyrocytes insulin induces protein accumulation but not DNA synthesis, while TSH does not increase protein accumulation but triggers DNA synthesis in the presence of insulin. We show here that EGF and phorbol myristate ester complement insulin action in the same way. HGF is the only factor activating both protein accumulation and DNA synthesis. The effects of insulin on protein accumulation and in permitting the TSH effect are reproduced by IGF-1 and are mediated, at least in part by the IGF-1 receptor. The concentration effect curves are similar for both effects. Similar results are obtained in human thyrocytes. They reflect true cell growth, as shown by increases in RNA content and cell size. Carbachol and fetal calf serum also stimulate protein synthesis and accumulation without triggering DNA synthesis, but they are not permissive for the mitogenic effects of TSH or of the general adenylate cyclase activator, forskolin. Moreover the mitogenic effect of TSH greatly decreased in cells deprived of insulin for 2 days although these cells remain hypertrophic. Hypertrophy may therefore be necessary for cell division, but it is not sufficient to permit it. Three different mechanisms can therefore be distinguished in the mitogenic action of TSH: (1) the increase of cell mass (hypertrophy) induced by insulin or IGF-1; (2) the permissive effect of insulin or IGF-1 on the mitogenic effect of TSH which may involve both the increase of cell mass and the induction of specific proteins such as cyclin D3 and (3) the mitogenic effect of the TSH cyclic AMP cascade proper.

Characterization of a thymidylate synthase (TS)-inducible cell line: a model system for studying sensitivity to TS- and non-TS-targeted chemotherapies

Thymidylate synthase (TS) is responsible for the de novo synthesis of thymidylate, which is required for DNA synthesis and repair and which is an important target for fluoropyrimidines such as 5-fluorouracil (5-FU), and antifolates such as Tomudex (TDX), ZD9331, and multitargeted antifolate (MTA). To study the importance of TS expression in determining resistance to these agents, we have developed an MDA435 breast cancer-derived cell line with tetracycline-regulated expression of TS termed MTS-5. We have demonstrated that inducible expression of TS increased the IC(50) dose of the TS-targeted therapeutic agents 5-FU, TDX, and ZD9331 by 2-, 9- and 24-fold respectively. An IC(50) dose for MTA was unobtainable when TS was overexpressed in these cells, which indicated that MTA toxicity is highly sensitive to increased TS expression levels. The growth inhibitory effects of the chemotherapeutic agents CPT-11, cisplatin, oxaliplatin, and Taxol were unaffected by TS up-regulation. Cell cycle analyses revealed that IC(50) doses of 5-FU, TDX and MTA caused an S-phase arrest in cells that did not overexpress TS, and this arrest was overcome when TS was up-regulated. Furthermore, the S-phase arrest was accompanied by 2- to 4-fold increased expression of the cell cycle regulatory genes cyclin E, cyclin A, and cyclin dependent kinase 2 (cdk2). These results indicate that acute increases in TS expression levels play a key role in determining cellular sensitivity to TS-directed chemotherapeutic drugs by modulating the degree of S-phase arrest caused by these agents. Moreover, CPT-11, cisplatin, oxaliplatin, and Taxol remain highly cytotoxic in cells that overexpress TS.

Nonapical and Cytoplasmic Expression of Interleukin-8, CXCR1, and CXCR2 Correlates with Cell Proliferation and Microvessel Density in Prostate Cancer

Purpose: We characterized interleukin-8 (IL-8) and IL-8 receptor expression (CXCR1 and CXCR2) in prostate cancer to address their significance to this disease. Experimental Design: Immunohistochemistry was conducted on 40 cases of human prostate biopsy containing histologically normal and neoplastic tissue, excised from patients with locally confined or invasive androgen-dependent prostate cancer, and 10 cases of transurethral resection of the prostate material from patients with androgen-independent disease. Results: Weak to moderate IL-8 expression was strictly localized to the apical membrane of normal prostate epithelium. In contrast, membranous expression of IL-8, CXCR1, and CXCR2 was nonapical in cancer cells of Gleason pattern 3 and 4, whereas circumferential expression was present in Gleason pattern 5 and androgen-independent prostate cancer. Each of IL-8, CXCR1, and CXCR2 were also increasingly localized to the cytoplasm of cancer cells in correlation with advancing stage of disease. Cytoplasmic expression (but not apical membrane expression) of IL-8 in Gleason pattern 3 and 4 cancer correlated with Ki-67 expression (R = 0.79; P <0.001), cyclin D1 expression (R = 0.79; P <0.001), and microvessel density (R = 0.81; P <0.001). In vitro studies on androgen-independent PC3 cells confirmed the mitogenic activity of IL-8, increasing the rate of cell proliferation through activation of both CXCR1 and CXCR2 receptors. Conclusions: We propose that the concurrent increase in IL-8 and IL-8 receptor expression in human prostate cancer induces autocrine signaling that may be functionally significant in initiating and promoting the progression of prostate cancer by underpinning cell proliferation and angiogenesis.

Downregulation of beta1,4-galactosyltransferase 1 inhibits CDK11(p58)-mediated apoptosis induced by cycloheximide.

Cyclin-dependent kinase 11 (CDK11; also named PITSLRE) is part of the large family of p34(cdc2)-related kinases whose functions appear to be linked with cell cycle progression, tumorigenesis, and apoptotic signaling. The mechanism that CDK11(p58) induces apoptosis is not clear. Some evidences suggested beta1,4-galactosyltransferase 1 (beta1,4-GT 1) might participate in apoptosis induced by CDK11(p58). In this study, we demonstrated that ectopically expressed beta1,4-GT 1 increased CDK11(p58)-mediated apoptosis induced by cycloheximide (CHX). In contrast, RNAi-mediated knockdown of beta1,4-GT 1 effectively inhibited apoptosis induced by CHX in CDK11(p58)-overexpressing cells. For example, the cell morphological and nuclear changes were reduced; the loss of cell viability was prevented and the number of cells in sub-G1 phase was decreased. Knock down of beta1,4-GT 1 also inhibited the release of cytochrome c from mitochondria and caspase-3 processing. Therefore, the cleavage of CDK11(p58) by caspase-3 was reduced. We proposed that beta1,4-GT 1 might contribute to the pro-apoptotic effect of CDK11(p58). This may represent a new mechanism of beta1,4-GT 1 in CHX-induced apoptosis of CDK11(p58)-overexpressing cells.