Effects of phorbol ester on carbachol-induced contraction in bovine ciliary muscle: possible involvement of protein kinase C

The aim of the research was to characterize muscarinic receptors of bovine ciliary muscle and to investigate the desensitization process. The role of protein kinase C was analyzed. The results show that muscarinic receptors of bovine ciliary muscle have the pharmacological characteristics of the M₃ subtype. Acute exposure to phorbol esters (1 μM phorbol 12,13-dibutyrate, PDB, or 0.1 μM phorbol 12-myristate 13-acetate, PMA, for 15 and 5 min, respectively) resulted in antagonism of muscarinic receptor-mediated contraction. Long-term pretreatment (18 h) with PMA to down-regulate protein kinase C resulted in potentiation of carbachol-induced contraction, reduction of agonist-induced desensitization and loss of phorbol ester-induced desensitization. Staurosporine (3 μM) and H₇ [1-(5-isoquinolinesulfonyl)-2-methyl-piperazine] (1 μM), protein kinase C inhibitors, produced a significant potentiation of the contractile effect of carbachol, reduced the desensitization produced by repeated addition of carbachol and suppressed that induced by phorbol esters. In vitro incubation with carbachol, PDB or PMA did not cause any modification of the binding of labeled [³H]quinuclidinyl benzilate. In vitro incubation with PDB and PMA produced, as expected, a significant translocation of protein kinase C from the cytosol to the membrane. The incubation of the ciliary muscle with carbachol, using the protocol of exposure that induced maximal desensitization of contractile responses, produced a significant redistribution of the enzyme from the cytosol to the membrane. These findings suggest that agonist-induced modulation of functional cholinergic sensitivity in ciliary muscle is correlated, at least partially, to the translocation of protein kinase C from the cytosol to the membrane. The desensitization by phorbol esters is completely due to protein kinase C activation; during the desensitization process, direct modification of the density and affinity of muscarinic receptors is not involved.     

谷氨酸脱氢酶,胆碱氧化酶和葡萄糖-6-磷酸酶对溶酶体中~(67)Ga积累的影响(英文)

目的:比较正常肝组织与肝癌AH 109A,吉田肉瘤中谷氨酸脱氢酶,胆碱氧化酶和葡萄糖-6-磷酸酶的活力对~(67)Ga摄取与积累的影响;方法:制备~(67)Ga枸橼酸溶液给大鼠静注后处死大鼠,制备亚细胞悬液,液闪计数器测定放射活度.结果:~(67)Ga的放射活性在正常肝组织溶酶体中(55％积聚)显著高于肝癌AH109A(32％积聚)和吉田肉瘤(18％)积聚.谷氨酸脱氢酶的活力在正常肝组织,肝癌和吉田肉瘤分别是1830±s 320 U·L~(-1),23±s 6 U·L~(-1)和7±s 2 U·L~(-1);胆碱氧化酶的活力分别是46±s 10 U·L~(-1),25.0±s 0.4 U·L~(-1),2.0±0.4 U·L~(-1);葡萄糖-6-磷酸酶活力分别是2550±s 180 U·L~(-1),84±s 14 U·L~(-1),78±s13 U·L~(-1).结论:正常肝组织中溶酶体酶活力很强,对~(67)Ga的积累起较大作用.癌变组织酶活力降低而作用减弱.吉田肉瘤细胞无肝细胞特点,其溶酶体对~(67)Ga积累作用不大.     

他汀类药物早期干预对急性冠脉综合征患者血脂、高敏C反应蛋白和纤维蛋白原的影响

目的 比较他汀类药物和阿司匹林联合用药与单用阿司匹林对急性冠脉综合征患者血脂、高敏C反应蛋白及纤维蛋白原的影响。方法 所有患者均在急性冠脉综合征发病后72h内开始接受药物治疗，他汀组(40例)应用他汀类药物加阿司匹林治疗8周，对照组(16例)单用阿司匹林治疗，观察两组总胆固醇、低密度脂蛋白胆固醇、高敏C反应蛋白和纤维蛋白原水平的变化。结果 治疗8周后，他汀组各指标显著降低，面对照组仅高敏C反应蛋白水平显著下降。他汀类药物降低高敏C反应蛋白、纤维蛋白原的程度与其降脂作用无关。结论 他汀类药物与阿司匹林联合用药降低高敏C反应蛋白和纤维蛋白原的作用可能优于单用阿司匹林，并与其抗炎作用有关。     

高压氧对家兔面神经撞击伤后面神经核内生长相关蛋白变化的影响(论著)

目的：研究面神经撞击伤后面神经核内生长相关蛋白（ＧＡＰ－４３）的变化及高压氧（ＨＢＯ）治疗的影响。方法：利用ＢＩＭ－Ⅱ型生物撞击机致家兔右侧面神经干损伤，应用ＳＰ免疫组化染色法观察损伤后１、３、７、１４和２１天（分别用ＨＢＯ治疗及不用ＨＢＯ为对照）面神经核内ＧＡＰ－４３的变化，并用电镜观察面神经干超微结构变化。结果：面神经损伤第１天，伤侧面神经核区ＧＡＰ－４３样免疫反应性神经元（ＧＬＩＮ）开始增多，第１４天其增多达到高峰，第２１天逐渐下降。ＨＢＯ治疗组伤后不同时期ＧＬＩＮ的增多明显高于对照组。伤后第２１天，与对照组相比ＨＢＯ组面神经干电镜观察可见较多的新生髓鞘。结论：ＨＢＯ治疗可使伤后面神经核内ＧＬＩＮ增多，因而有助于面神经损伤后的修复与再生。     

Extraction of Serum Proteins Adsorbed on the Surface of Dialysis Membranes

Abstract: Extraction of adsorbed proteins from dialysis membranes that had been used during actual hemodialysis procedures was performed. The condition of extraction with SDS plus 2–mercaptoethanol at 95°C is more efficient than with only PBS or with SDS solution without 2–mercaptoethanol at 37°C.     

构建玻片蛋白质芯片的实验研究

目的:探讨玻片蛋白质芯片的构建方法及其中间操作条件的选择与优化。方法:利用生物芯片点样仪将超微量蛋白质点到经特殊处理的玻璃片表面,然后选用不同的化学试剂对玻片背景进行封闭;再用荧光素标记的蛋白质与点样蛋白杂交;最后用生物芯片分析仪扫描成像并进行分析,比较不同条件下芯片的显示效果。结果:蛋白质样品与片基稳定结合,并保持原活性状态;封闭试剂选用酪蛋白或明胶效果较佳;点样探针与其特异蛋白质抗体可稳定结合,结合效果与点样蛋白浓度在一定范围内呈正相关;在1.6 cm×1.6 cm的玻璃片面积上,构建了36×36=1269点的蛋白质芯片。结论:本实验条件下,蛋白质抗原或抗体可以稳定地固定于经过处理的玻璃片表面.制成免疫型蛋白芯片,并可通过随后的抗原抗体反应与荧光素标记的相应抗体或抗原结合,用生物芯片分析仪可对其荧光信号进行检测分析。     

肠溶阿斯匹林剂量和血阿斯匹林及水杨酸盐浓度与药效关系探讨

本文探讨了ＡＳＡ剂量与药效的关系及血ＡＳＡ、ＳＡ药物浓度监测的临床意义，结果表明：（１）ＡＳＡ剂量与６－ｋｅｔｏ－ＰＧＦ_（１α）抑制率呈正相关（Ｐ＜０．０１），而与ＴＸＢ_２及ＰＡｇＲ抑制率无相关性（Ｐ＞０．１）；（２）本文采用ＨＰＬＣ内标法同时测定血ＡＳＡ和ＳＡ浓度，结果准确，方法简便；（３）当口服小剂量ＡＳＡ防治ＣＩ时，监测血ＡＳＡ和ＳＡ以调整ＡＳＡ用药剂量的临床价值并非十分重要。     

Study on lymphocyte activation and proliferation induced by anti-CD3 McAb

Summary T cell activation and proliferation via CD₃-TCR complex were investigated by lymphocyte DNA synthesis in vitro. Several interfering factors were also discussed. The result indicated that lymphocyte activation and proliferation are calciumdependent. A rise of cytoplasmic free Ca²⁺ quickly following activation with CD₃ McAb is mainly due to intracellular mobilization of Ca²⁺, while lymphocyte proliferation needs both intracellular mobilization of Ca²⁺ as well as influx of extracellular Ca²⁺. It was confirmed that CTX sensitive G protein plays a role in regulating T cell proliferation by pretreatment with CTX suppressing lymphocyte³H-TdR incorporation obviously. PLC and PKC inhibitor neomycin and P. S. S could also decrease T cell proliferation.     

The influence of anaesthetic techniques and type of delivery on peripartum serum interleukin-6 concentrations

Background: Interleukin-6 is a pleiotropic cytokine with a wide range of physiological activities. It plays an important role in the immuno-neuro-humoral axis during stress and surgery.
Methods: Serum interleukin-6 in parturients was measured on hospital admission, immediately after birth and 12 and 24 hours later. All parturients had uncomplicated pregnancies, and delivered vaginally without (n=31) or with (n=20) epidural analgesia, or underwent Caesarean section under epidural (n=20) or general (n=10) anaesthesia.
Results: Serum interleukin-6 assayed immediately following Caesarean section was low, but peaked 12 hours later, irrespective of the anaesthetic technique or other foetomaternal characteristics. Patients who delivered vaginally showed the highest interleukin-6 levels immediately after delivery. These were positively correlated with serum interleukin-6 on admission and duration of labour. Serum interleukin-6 was significantly higher in parturients who had epidural analgesia, and was significantly lower in those receiving intravaginal prostaglandins compared to those without prostaglandins.
Conclusion: The interleukin-6 response after Caesarean section can be explained by a generalized acute phase response to surgery, with no anaesthetic, maternal or neonatal interference. The rapid increase in peripartum serum interleukin-6 levels after vaginal delivery reflects, in part, cervical ripening or labour, their physiological triggers and psychological or physical stress. Regional anaesthesia, duration of labour and exogenous prostaglandin administration can modulate the peripartum interleukin-6 response and subsequently the physiological effects of this cytokine.