黄芪丹参复方成分对一氧化氮合成阻滞孕鼠模型血浆IL-1、IL-10变化的影响

目的探讨黄芪丹参复方成分提高胎盘血供的分子机制，为临床有效防治胎盘血供不足所致妊娠并发症提供思路。方法提取分离黄芪、丹参复方成分，运用NOS（一氧化氮合酶）阻滞剂L－精氨酸甲酯（L—NAME）建立一氧化氮合成阻滞大鼠模型，ELISA法检测妊娠18d大鼠血浆IL－1、IL－10水平。以及胎盘超微形态学变化，大鼠血压、尿蛋白变化，以及仔鼠重、肝重、脑重、胎盘重等。结果一氧化氮合成阻滞模型组IL－1含量明显高于空白组（P〈0．01），经黄芪丹参注射液治疗后，血浆IL－1水平比模型组降低（P〈0．05）；模型组IL－10含量较空白组低（P〈0．01），中药组血浆IL－10水平高于模型组（P〈0．05）；胎盘形态学、血压、尿蛋白以及仔重、仔肝重、脑重等均有显著差异（P〈0．05，P〈0．01）。结论黄芪丹参复方成分可能对妊娠早期母－胎界面免疫平衡具有调控作用，通过促进局部生理抑制性免疫反应增强和杀伤、排斥免疫反应减弱，从而有利于母胎循环构建，维持胎盘血液供应。     

缺血再灌流肾组织内皮素—1动态变化的实验研究

在大鼠肾缺血６０分钟再灌注的模型上观察不同时相肾静脉血、肾皮质、外髓和内髓的内皮素１（ＥＴ１）浓度变化，肾组织ＥＴ１光镜和电镜免疫组织化学变化。结果发现：缺血再灌流肾组织ＥＴ１基因表达及分泌明显增强，主要分布在血管内皮细胞及平滑肌细胞、系膜细胞、肾小管上皮细胞。其分布特点与细胞类型和活性有关。本实验结果提示了缺血再灌注肾内ＥＴ１的变化规律。     

Expression of human T-cell lymphotropic virus type-1 gene products in the short-term cultured skin tissues of an adult T-cell leukemia/lymphoma patient with cutaneous manifestations.

Adult T-cell leukemia/lymphoma (ATLL) is recognized as a disease etiologically associated with human T lymphotropic virus type-1 (HTLV-1) infection, but, neither viral replication nor specific virus antigen expression have been detected on ATLL cells distributed in organs, including skin. To examine the latent expression of HTLV-1 in the cutaneous lesions of ATLL patients, we cultured the lesional skin tissues in vitro and applied immunofluorescence staining with mouse monoclonal antibodies Lt-4, GIN-14, and F10, which react with p40tax, p19 and gp21, respectively. We recognized HTLV-1 specific antigens on clustered ATLL cells only in the deeper dermis of the skin after 24 hrs cultivation of the lesional skin tissue from an ATLL patient in RPMI-1640 medium supplemented with 20% fetal calf serum. In the electron microscope, we observed HTLV-1 like particles, 80-140 nm in diameter with envelope and core structures, in the same tissue specimen. These findings suggest that HTLV-1 gene products may be expressed in the skin lesions of ATLL patients and involved in the pathogenesis of skin eruptions in cutaneous type ATLLs. To our knowledge, this is the first report that envisages the potency of intracutaneous HTLV-1 expression in vivo.     

Serum markers for fibrosis and plasma transforming growth factor-β1 in patients with hepatocellular carcinoma in comparison with patients with liver cirrhosis

In order to elucidate collagen metabolism in hepatocellular carcinoma (HCC) tissue, we compared levels of different potential markers of collagen metabolism and plasma transforming growth factor-β₁ in patients with HCC and in patients with liver cirrhosis. Serum levels of prolyl hydroxylase and the tissue inhibitor of metalloproteinase-1 in patients with HCC were significantly higher than those in patients with liver cirrhosis and increased with the size of the HCC tumour, whereas the serum levels of procollagen type III propeptide and type IV collagen 7S domain were similar in the two groups. In HCC, the increased plasma transforming growth factor-β₁ levels were closely correlated with serum levels of prolyl hydroxylase and the tissue inhibitor of metalloproteinase-1. These findings suggest that, in HCC tissue, the intracellular biosynthesis of collagen is enhanced, whereas the secretion of procollagen is disturbed and the degradation of collagen is suppressed by the excess production of the tissue inhibitor of metalloproteinase-1. The results also suggest that plasma transforming growth factor-β₁ plays an important role in the altered metabolism of collagen in HCC.     

HIGH-LEVEL EXPRESSION OF RATD1A DOPAMINE RECEPTOR cDNA IN MOUSE FIBROBLASTLTK- CELLS BY n-BUTYRATE

1. In orefer to develop a simple, efficient system for the highlvel expression of dopamine recetors in eukaryotic cells, we have studied the effects of n-butyrate on the expression of rat D_1A dopamine receptor cDNA in mouse fibroblast LTK- cells as compared with those of n-butyrate on endogenous D₁ receptor levles in opossum kidney cells.
2. In the transfected LTK- cell mebranes with pRc/CMVD_1A receptor cDNA, a selective D₁ dopamine antagonist, [³H]-SCH 23390, exhibited a K_d of 0.9 ± 0.1 nmol/L and a B_max of 0.35 ± 0.05 pmol/mg protien (n = 5).
3. Addtion of n-butrate (2–10 mmol/L) to the culture medium for 48h dose-dependently increased the D_1A receptor level up to 1.5 ± 0.3 pmol/mg protien (n = 7), although the K_d values were not affected. The increase in receptor level was accompanined by an elevation of selective D₁ agoinist-induced adenyly cyclase activity.
4. In contrast, n-butyrate treatment (2–10 mmol/L) did not affect either endogenous D₁ receptor levels or fendoldopmainduced adenylyl cyclase activity in possum kidney cells.
5. These results sugges n-butyrate is a sueful tool for obtaining high-level expression of D_1A dopamine receptor cDNA in mouse fibroblast LTK- cells.     

Patch clamp studies on the kinetics and selectivity of N-methyl-d-aspartate receptor antagonism by memantine (1-amino-3,5-dimethyladamantan)

Memantine (1-amino-3,5-dimethyladamantan) was tested as an antagonist of N-methyl-d-aspartate (NMDA) receptors on cultured superior collicular and hippocampal neurones using the patch clamp technique and its actions were compared to those of Mg²⁺ ions, ketamine, dextrorphan, dextromethorphan, phencyclidine and dizocilpine (MK-801). Memantine (2–33 μM) concentration-dependently antagonized responses to NMDA 100 μM with an IC₅₀ of 2.92 ± 0.05 μM. In contrast, current responses to (S)-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (l-AMPA 50–100 μM) and γ-amino butyric acid (GABA 10 μM) were unaffected by Memantine 8 μM. Memantine 8 μM caused a non-parallel shift of the NMDA concentration-response curve to the right in a manner indicative of uncompetitive open channel block. The effects of memantine were similar to ketamine in that both antagonists were weakly use- and strongly voltage-dependent. In contrast, MK-801, phencyclidine and dextrorphan showed much slower kinetics that was reflected in their marked use- and weaker voltage-dependency. The antagonistic effects of memantine were not reversed by increasing concentrations of glycine (0.1–100 μM) ruling out the possibility of an interaction of memantine with the strychnine-insensitive glycine modulatory site associated with the NMDA receptor-channel complex. Memantine (1–100 μM) also selectively antagonized responses to NMDA (40 μM) in the cortical wedge preparation with IC₅₀ of 12.9 ± 1.5 μM.     

Pyrroloquinolones and pyrazoloquinolones as potential antibacterial agents. Synthesis and antibacterial activity

Diethyl 1-cyclopropyl-5,7,8-trifluoro-1,4-dihydro-4-oxoquinoline-3,6-dicarboxylate 4 as a key-intermediate was synthesized via the Dieckmann reaction. The reaction of 4 with nucleophiles proceeded regioselectively at C-5. Facile cyclization between the C-5 and C-6 side chains of the resulting products gave novel pyrroloquinolones 10 and 12 and pyrazoloquinolones 15. They were converted into a series of cyclic amino-substituted pyrroloquinolones 17–21 and pyrazoloquinolones 22–24, and their in vitro antibacterial activities were tested. 1H-Pyrrolo[2,3-f]quinolone 17a and 2H-pyrrolo[3,4-f]quinolone 21a exhibited a potent in vitro antibacterial activity.     

细小病毒H－1对正常仓鼠肝细胞DNA周期动力学的影响

健康成年金黄仓鼠腹腔内接种ＰＶＨ－１（５×１０６ｐｆｕ），ＦＣＭ动态检测肝细胞ＤＮＡ周期动力学变化，结果显示：ＰＶＨ－１接种后处于Ｇ０＋Ｇ１期的肝细胞与对照组动物相比无显著差异。Ｓ期肝细胞在第３天显著高于对照组动物（Ｐ＜０．０５），且在第７天时仍呈上升趋势，第１４天到３０天趋于正常水平。Ｇ２＋Ｍ期肝细胞在ＰＶＨ－１接种后第３０天升高（Ｐ＜０．０５）。结果提示，动物在ＰＶＨ－１接种后第３到第７天是病毒在肝细胞中的复制高峰，并阻滞仓鼠处于Ｓ期肝细胞分裂进入Ｇ２＋Ｍ期，但是这种阻滞作用较短暂、并可逆，在ＰＶＨ－１接种后１４～３０天自然消失。     

5-HT1A receptor agonists: recent developments and controversial issues

During the last decade, serotonin (5-HT)_1A receptors have been a major target for neurobiological research and drug development. 5-HT_1A receptors have been cloned and a variety of selective agonists, such as the aminotetraline 8-OH-DPAT and the pyrimidinylpiperazine ipsapirone, have become available. Demonstrations of apparent intrinsic activity of these ligands at 5-HT_1A receptors, however, depend highly on the particular assay system. This may be due to the possible existence of receptor subtypes and to assay (or brain region)-dependent differences in receptor reserve and the nature of receptor-effector coupling. Nevertheless, the apparent intrinsic activity of 8-OH-DPAT seems to be higher (although possibly not yet maximal) than that of the pyrimidinylpiperazines. In the brain, 5-HT_1A receptors are located presynaptically as somatodendritic receptors on 5-HT neurons and postsynaptically in particular limbic and cortical regions. Although it is generally accepted that presynaptic 5-HT_1A receptors control 5-HT neuronal activity, recent evidence suggests an additional role of postsynaptic 5-HT_1A receptors in cortex as part of a negative feedback loop. Anxiolytic and antidepressive properties of selective 5-HT_1A receptor agonists have now been confirmed by clinical studies. Although it is well established that the latter properties depend on theagonistic activity of these compounds, theoptimal level of intrinsic activity is still a matter of debate and may be dependent on the clinical indication. Such compounds may also have antiaggressive effects, and possibly anticraving effects (manifested by their alcohol intake-reducing effects in dependent animals), but the specificity of these so-called anti-impulsivity effects is still controversial and not yet tested clinically. Anticataleptic, antiemetic and neuroprotective properties have been demonstrated in different species. Behavioral studies on the mechanisms underlying the anxiolytic and antidepressive effects have examined the relative contribution of pre-and postsynaptic 5-HT_1A receptors by means of local cerebral application and lesion techniques. Most evidence points towards a critical involvement of presynaptic receptors in the anxiolytic effects of 5-HT_1A receptor agonists (although a possible contribution of postsynaptic receptors cannot be excluded). With regard to the antidepressive properties, a case can be made for the reverse; i.e., a strong involvement of postsynaptic receptors and a questionable contribution of presynaptic receptors. However, as the therapeutic effects of those 5-HT_1A receptor (partial) agonists which have been tested clinically require repeated administration, attention has been directed increasingly towards chronic studies. These studies have shown that a number of electrophysiological, biochemical, behavioral and endocrinological 5-HT_1A receptor-related events adapt differentially to repeated or sustained administration. Thus, several hypotheses accounting for the delayed onset of action have been advanced. Among these, time-dependent downregulation /desensitization of eitherpre- orpostsynaptic 5-HT_1A receptors, or cortical 5-HT₂ receptors have received much attention. However, these hypotheses have their weaknesses, and it is argued thatfunctional sensitization of particular postsynaptic 5-HT_1A receptor-mediated events remains a valuable alternate hypothesis. Basic research on the role of 5-HT_1A receptors in psychopathology and in the therapeutic effects of clinically effective therapeutics, as well as on the mechanism of action of 5-HT_1A receptor ligands, will enable rational design of ligands with particular profiles of intrinsic activity at different 5-HT_1A receptor populations, and may contribute to a more efficient treatment of a multiplicity of brain disorders.     

P物质及其受体在慢性胰腺炎组织的表达和临床意义

目的 了解P物质及其受体NK－1R在正常胰腺和慢性胰腺炎中的表达，探讨它们与慢性胰腺炎时疼痛发生的关系。方法 25个慢性胰腺炎标本取自慢性胰腺炎手术，男性18例，女性7例，正常胰腺组织取自20个器官捐献者，男性11例，女性9例。应用实时定量逆转录整合酶链反应（RT－PCR）技术，检测正常胰腺和慢性胰腺炎组织中P物质和NK－1R的mRNA水平，应用Western blot技术检测NK－1R的蛋白水平，应用免疫组织化学方法进行NK－1R的组织学定位。将P物质及其受体NK－1R mRNA水平与疼痛的程度，发作频率和病程进行分析，寻找其中的相关性。结果 与正常胰腺相比，慢性胰腺炎组织中P物质和NK－1R mRNA和蛋白都过度表达，NK－1RmRNA水平与疼痛的程度，发作频率和病程明显相关。免疫组化检查显示，NK－1R的表达主要位于胰腺腺泡，胰腺导管，神经纤维和炎性细胞。结论 慢性胰腺炎组织中P物质和NK－1R的表达水平都明显上调，扰乱了神经激肽的作用环节，NK－1R的过度表达与疼痛发生有关。