高效液相法测定金樱子中三萜酸类成分的含量

定量测定不同产地金樱子中2α,3α,19α,23-四羟基乌苏-12-烯-28-羧酸的含量.采用高效液相法,色谱柱:Kromasil-C18(4.6 mm×250 mm,5 μm)柱,大连依利特公司;检测波长203 nm,柱温:35 ℃;流动相:乙腈-水(35:65),流速1 mL/min.结果表示所测成分与其他组分具有良好的分离度,线性范围为0.4～10 μg, 加样回收率为102.5%,RSD=2.66%.说明不同产地的三萜类有机酸的含量差别很大.本方法操作简单,结果准确,可用于含2α,3α,19α,23-四羟基乌苏-12-烯-28-羧酸的药物的含量测定.     

Prospect for enzyme therapy in glycogenosis II variants: a study on cultured muscle cells

Summary Impairment of skeletal muscle function is the common feature of distinct clinical forms of glycogenosis type II. In the present study, muscle cultures from different patients were used to investigate the cause of clinical heterogeneity and the feasibility of enzyme replacement therapy. The activity of acid -glucosidase appears to be the primary factor in determining the extent of lysosomal glycogen storage in muscle, and thereby the clinical severity of the disease. Neutral -glucosidases do not seem influencial. Correction of the enzymatic defect was achieved in skeletal muscle cultures from patients by administration of a high-uptake form of acid -glucosidase, purified from human urine. The enzyme reaches the lysosomes, including the glycogen storage vacuoles, and the lysosomal glycogen content is reduced to control level. In normal muscle cells 20% of the total cellular glycogen pool is segregated in lysosomal compartments. This percentage is higher than in fibroblasts, which may partly explain why muscles are more prone to store glycogen. The relevance of this study for enzyme therapy is discussed.     

Autonomic control of acid phosphatase exocrine secretion by the rat prostate

Summary In vivo prostatic secretion was collected from retired breeder Sprague Dawley rats using a method for isolated perfusion of the rat prostatic urethra. Enzymatic acid phosphatase determination was performed on the collected effluent. Control acid phosphatase secretion was 24.2±2.7 nm over 30 minutes. Intravenous phenylephrine 5 mg/kg stimulated a 10 fold increase in acid phosphatase secretion. The secretion seen with phenylephrine was dose dependent and could be blocked with prazosin, but not yohimbine, atropine, or propranolol. Intravenous -adrenergic agonist isoproterenol caused no increase in the secretion of rat prostatic acid phosphatese. Intravenous administration of the cholinergic agonist pilocarpine also resulted in a dose dependent rise in acid phosphatase secretion. The stimulation seen could be blocked by atropine but not phentolamine or propranolol. The stimulation of acid phosphatase secretion seen with ₁ adrenergic or cholinergic agonists was not additive. Intravenous vasoactive intestinal peptide did not stimulate acid phosphatase secretion nor did it augment the secretion induced by ₁ adrenergic or cholinergic agonists. Release of acid phosphatase into rat prostatic exocrine secretion is under both ₁ adrenergic and cholinergic control.Supported by the US Veterans Administration     

The anaerobic recovery of frog muscle

Summary The muscle may undergo a partial recovery of its high energy phosphate stores in the absence of oxygen by the way of glycolysis (anaerobic recovery). This process has been studied in 41 pairs of frog gastrocnemii at different degrees of exhaustion induced by variable trains of supramaximal stimuli. Anaerobic recovery appears to be inadequate to replenish the fraction of muscle high energy phosphate stores (GP=ATP+PC) split as a consequence of the stimulation. The maximal amount of recovery (on the average about 5 Moles of GP per gram of fresh tissue) occurs when the muscle resting stores have been reduced to about 50%. This limitation in the extent of recovery is not a consequence of a reduced availability of glycogen but it is possibly related to the production of some metabolic intermediate, limiting the rate of the glycolytic sequence, likely the accumulation of lactic acid in the fiber. The time course of the anaerobic recovery process is characterized by at1/2 of about 2 min. The efficiency of the process, i.e. the number of the high energy phosphate bonds resynthesized by one Mole of lactic acid, appears to vary between 1.5 and 1.8, being of the same order of magnitude as the GP/L.A. ratio obtained from muscle extracts.     

Utilization of carbohydrates and free fatty acids by the gastrocnemius of the dog during long lasting rhythmical exercise

Summary Utilization of carbohydrates and free fatty acids (FFA) has been investigated in gastrocnemii of dogs during long lasting isotonic rhythmical exercise induced by supramaximal stimulation of the sciatic nerve. Uptake or output of gases and substrates was determined according to the Fick principle. The first measurements were done at about 2 min after the beginning of work when blood flow has reached a steady state, and the latest at about 100 min after the beginning of exercise.During the first 7 min when the work performed exceeded 5 kg/100g×min and O₂ consumption exceeded 11 ml/100g×min, uptake of arterial glucose and FFA was low, accounting for less than 40% of the total O₂ consumption. Since the RQ values at the same time were about 1.0, glycogen must have been oxidized as the major aerobic energy source.About 13 min after the beginning of exercise, the work the muscles could perform declined to about half of the initial value and remained so for the following 90 min. During this time the oxygen extraction ratio of FFA was about 50% and of arterial glucose was 40–50%, while the RQ value was about 0.8.During initial strong exercise an output of lactic acid (LA) of about 10 mg/100 g×min was measured. With the decrease of work as a consequence of fatigue, LA output became negligible, and in many experiments small amounts of LA were taken up by the working gastrocnemii.It is concluded that glycogen is the major aerobic energy source for strong muscular exercise which cannot be substituted for by the oxidation of arterial glucose or FFA.Supported by the Deutsche Forschungsgemeinschaft.     

The rate of lactic acid removal in relation to different baselines of recovery exercise

Summary During strenuous exercise lactic acid (LA) appears in the blood as a result of anaerobic metabolism. The rate at which this LA was removed from the blood after exercise was seen to increase to a maximum with a certain level of post exercise activity. This maximum rate of removal of LA appears to be at approximately 40% of the individual's maximum oxygen uptake when the exercise is performed on a bicycle ergometer.     

白细胞介素2和黄芪多糖对小鼠NK细胞作用的细胞化学和超微结构观察

本文报道在白细胞介素2(IL-2)及黄芪多糖(PA)作用下,小鼠NK细胞的细胞化学和超微结构变化。实验表明,NK细胞酸性磷酸酶、酸性醋酸酯酶及氯醋酸AS-D酯酶均为阳性。经IL-2和PA的处理,前两种酶的活性增加,阳性细胞百分率增高,并与NK活性相关。IL-2培养后的NK细胞超微结构表现为胞体变大,表面不规则,胞质丰富,细胞器增多等特点。     

新城疫病毒pIRHN核酸疫苗构建和表达及对肿瘤细胞的影响

目的：研究NDV HN基因抗肿瘤作用及其可能机制。方法：以 pIRES1neo为表达载体构建了 NDV HN基因的pIRHN核酸疫苗，在体外转染HeLa细胞，用间接免疫荧光和Western blot检测pIRHN在真核细胞中表达状况，用荧光显微镜、DNA琼脂糖电泳及TUNEL染色等方法，检测HN基因导致细胞死亡的类型；用3，5-二羟基甲苯法测定HeLa细胞唾液酸含量的变化。结果：pIRHN 转染HeLa细胞后，能够在真核细胞中表达，能促进肿瘤细胞死亡，其死亡方式主要以诱导细胞凋亡为主，pIRHN使 HeLa细胞唾液酸含量减少。结论：用 NDV HN基因所构建的核酸疫苗能够在真核细胞中高效表达，表达的 HN蛋白主要位于胞膜，胞浆中亦有 HN蛋白表达；pIRHN具有抗肿瘤作用，可能通过其表达产物与肿瘤细胞唾液酸受体的相互作用，发挥其抗肿瘤作用。本实验为迸一步阐明NDV抗肿瘤作用机制提供了理论依据。     

Ventral medulla pHi measured in vivo by 31P NMR is not regulated during hypercapnia in anesthetized rat

Chemoreceptors in the ventral medulla contribute to the respiratory response to hypercapnia. Do they ‘sense’ intracellular pH (pH_i)? We measured pH_i in the ventral medulla or cortex (control) using ³¹P-NMR obtained via a novel 3×5 mm² surface coil in anesthetized rats breathing air or 7% CO₂. During air breathing over 240 min, pH_i decreased slightly from 7.13±0.02 to 7.05±0.02 (SEM; n=5; 2 cortex, 3 ventral medulla). During 180 min of hypercapnia, cortical pH_i (n=4) decreased from 7.17±0.02 to 6.87±0.01 by 90 min and recovered by 150 min. Ventral medulla pH_i showed no such regulation. It decreased from 7.11±0.02 to 6.88±0.02 at 90 min and recovered only after cessation of hypercapnia (n=5), results consistent with pH_i being the chemoreceptor stimulus. However, non-chemoreceptor neurons that contribute to our medullary NMR signal also do not appear to regulate pH_i in vitro. Regional differences in pH_i regulation between cortex and ventral medulla may be due to both chemosensitive and non-chemosensitive neurons.     

Action du clopamide et de l'acide ethacrynique sur la sécrétion de rénine chez le chien

Summary Renin secretion was studied in anesthetized dogs (Pentobarbital), after an injection of 5 mg/kg i.v. clopamide (10 animals) or ethacrynic acid (10 animals). Renin activity in venous renal blood (Boucher's method), renal blood flow (electromagnetic flowmeter), blood pressure, diuresis and natriuresis have been measured simultaneously. Basic renin secretion was 22,4±8,2 ng/g of kidney/min. During the 8 min following the ethacrynic acid injection renin secretion increased to 74,4±41,1 ng/g of kidney/min (p<0.005). Within the same time it dropped to 11.1±6.8 ng/g of kidney/min (p<0.005) after clopamide.These variations of renin secretion are related to the diuretic's action on the nephron, and depend on an endorenal mechanism. Ethacrynic acid suppresses the corticopapillary gradient while clopamide does not. We have not found any correlation between renin secretion and either natriuresis (mg/min) or diuresis (ml/min). But there is a connection (r=0.7367) between renin secretion after ethacrynic acid injection and sodium concentration in urine.