Effect of Co-administration of Morphine and Cholinergic Antagonists on Y-maze Spatial Recognition Memory Retrieval and Locomotor Activity in Mice

The interaction of morphine and cholinergic system was shown in previous studies. In the present study, we investigated whether morphine would interact with the cholinergic antagonists, scopolamine and atropine in a Y-maze spatial recognition memory. Pre-test treatments of morphine （5, 1.5, 0.5 mg/kg）, scopolamine （1, 0.1 mg/kg）, atropine （0.5, 0.1 mg/kg） were used in the experiments, relatively high or low doses were paired respectively as co-administration measures. The results showed that co-administration of morphine 0.Smg/kg ~ scopolamine 0.1 mg/kg and morphine 0.5 mg/kg ＋ atropine 0.1 mg/kg disturbed the inspective exploratory behavior （percent of arm duration） but not the inquisitive behavior （percent of arm visits） of the spatial memory retrieval, while the drugs didn＇t cause amnesia when single administered of the concerned low doses. Distinct interaction was found between scopolamine and morphine on increasing locomotor activity.

Role of intravenously administered hyoscine butyl bromide in retrograde terminal ileoscopy: A randomized, double-blinded, placebo-controlled trial

AIM： To evaluated the role of hyoscine butyl bromide in facilitating retrograde ileoscopy. METHODS： Retrograde terminal ileoscopy was attempted in 200 consecutive patients undergoing colonoscopy. After intubation of the cecum and visualization of the ileocecal valve, butyl bromide injection or normal saline was given intravenously to the patients in a double blind random fashion. The pulse rate and oxygen saturation were measured continuously. After completion of the procedure, endoscopists were then asked to score the ease of intubation and the ease of visualization of the terminal ileum on a visual scale of 1 to 10. The patients were also asked to score the pain after receiving hyoscine butyl bromide injection on a score of 1 to 10. RESULTS： Terminal ileoscopy could be performed in 188 patients. The mean （SD） visual analogue score for the ease of intubation of the cecum was 7.4 （0.65） in the injection group and 5.9 （0.8） in the placebo group （P 〈 0.001）. The mean （SD） length of ileum visualized in the injection group was 14.4 （3.3） cm and 10.4 （2.7） cm in the placebo group （P 〈 0.001）. The mean （SD） visual analogue score for ease of visualization of the terminal ileum was 7.5 （0.69） in the injection group and 5.9 （0.7） in the placebo group （P 〈 0.001）. The pain score experienced by the patients was 6.5 （0.7） in the injection group and 6.7 （0.69） in the placebo group （P 〈 0.008）. Although the pulse rate increased significantly in patients receiving the drug, no statistically significant difference was noted in the oxygen saturation between the two groups either before or after administration of the drug. No complications were observed in either of the groups. CONCLUSION： Hyoscine butyl bromide injection is a useful adjunct in helping the intubation and visualization of terminal ileum during colonoscopy.

液液萃取-气相色谱-质谱联用法同时测定曼陀罗中的消旋山莨菪碱、东莨菪碱与(-)-莨菪碱

目的建立一种气相色谱-质谱联用法同时测定消旋山莨菪碱、东莨菪碱与（-）-莨菪碱的方法。方法曼陀罗经2mol／L盐酸超声提取30min后过滤，滤液用浓氨水调节pH≈9后用三氯甲烷提取3次，挥干定容后上机测定。结果消旋山莨菪碱、东茛菪碱与（一）一莨菪碱在0．1～0．5mg／ml质量浓度范围内呈良好的线性关系，相关系数（r^2）为0．9981～0．9995，在0．2～0．4mg／ml两个浓度水平的加标回收率为76．7％～119．0％，RSD为0．8％～7．3％，检出限为0．1～1．0μg／kg。结论本方法简单、快速、可靠，能够满足曼陀罗中消旋山莨菪碱、东莨菪碱与（-）-莨菪碱的检测分析。

Potencies of Four Stereoisomers of Anisodamine on Muscarinic Receptor

用[3~H]-QNB配体受体结合实验,抗Ach对豚鼠回肠纵肌条收缩效应,抗毛果芸香碱流涎及扩瞳效应为指标,比较山莨菪碱四个立体异构体(A.6S,2′S;B.6S,2′R;C.6R,2′R;D.6R,2′S)与M受体相互作用的构效关系,结果A,B,C,D四个异构体效应分别为:(1)受体结合实验K_i(M)2.69×10^(-7),1.43×10^(-7),1.63×10^(-5),2.15×10^(-6);(2)抗Ach豚鼠回肠纵条收缩:K_6(M)2.98×10^(-8);1.38×10^(-8);1.52×10~5;1.0l×10^(-7);(3)扩瞳:ED_4(mol/kg)1.6×10^(-6),1.9×10^(-6),5.1×10^(-5),3.6×10^(-5);(4)抗毛果芸香碱流涎:ED_(50)(mol/kg)1.5×10^(-6);1.3×10^(-6);2.6×10^(-5);3.1×10^(-5)。以上结果表明四个异构体效应强度为6S,2′R≌6S,2′S>>6R,2′S>6R,2′R。

The mixture of procyanidins extracted from the lotus seed pod and bilobalide ameliorates scopolamine-induced memory impairment in mice

Objective To study the co-effect of procyanidins extracted from the lotus seed pod （LSPC） and bilobalide （BIL） on ameliorating scopolamine-induced learning and memory impairment in young mice. Methods Fifty male Kunming mice with similar learning and memory capabilities were selected by Morris water maze test and were randomized into 5 groups （n=10 in each group）： control group, scopolamine group, L-（LSPC＋BIL） group （50 mg/kg LSPC＋10 mg/kg BIL）, M-（LSPC＋BIL） group （100 mg/kg LSPC＋20 mg/kg BIL）, H-（LSPC＋BIL） group （150 mg/kg LSPC＋30 mg/kg BIL）. Scopolamine model with impaired learning and memory was established by scopolamine treatment （1 mg/kg）, and after 10 min mice were tested. In L-, M-, and H- （LSPC＋BIL） groups, mice were treated with LSPC and BIL ig. for 30 days, while mice in the other 2 groups were treated with normal saline ig. instead. After the 30-day＇s treatment, the co-effect of LSPC and BIL on learning and memory was tested by Morris water maze and the step-down avoidance tests. Results The memory impairment caused by scopolamine in young mice could be ameliorated by co-treatment of LSPC and BIL, as indicated by significantly shorter escape latency and swimming distance in the Morris water maze test, when compared with those in the scopolamine group. In the step-down avoidance test, mice in all the 3 dose groups showed significantly smaller number of errors and longer latency than mice in the scopolamine group did. Conclusion Co-treatment of LSPE and BIL can ameliorate scopolamine-induced learning and memory impairment in young mice.

Cloning of H6H Gene from Atropa belladonna and Construction of the Efficient Plant Expression Vector

[Objective] The aim was to clone H6H gene from Atropa belladonna and construct an efficient plant expression vector.[Method] The coding sequence of H6H（Hyoscyamine 6β-hydroxylase）was cloned from Atropa belladonna with RT-PCR.Then,the sequence was subcloned into the reconstructed plant binary expression vector p2301 to construct the recombinant vector p2301-H6H,which was then introduced into Agrobacterium tumefaciens strain LBA4404 and Agrobacterium rhizogenes strain C58C1,respectively.[Result] The engineering bacteria p2301-H6H-LBA4404 and p2301-H6H-C58C1 which could be directly used in genetic improvement were obtained.[Conclusion] The present research provided basis for the increasing of alkaloid content of Atropa belladonna by plant genetic engineering technology.

The effects of anisodamine and dobutamine on gut mucosal blood flow during gut ischemia/reperfusion

AIM: To determine if anisodamine is able to augment mucosal perfusion during gut I/R ischemia-reperfusion. METHODS: A jejunal sac was formed in Sprague Dawley rat. A Laser Doppler probe and a tonometer were inserted into the sac which was filled with saline. The superior mesenteric artery was occluded (SMAO)for 60 minutes followed by 90 minutes of reperfusion. At the end of 60 minutes of SMAO, either 0.2 mg/kg of anisodmine or dobutamine was injected into the jejunal sac. Laser Doppler mucosal blood flow and regional PCO2 (PrCO2)measurements were made. RESULTS: Mucosal blood flow was significantly increased at 30,60 and 90 minutes of reperfusion (R30, R60, R90) when intraluminal anisodamine or dobutamine was present compared to intraluminal saline only(44+/-3.3% or 48+/-4.1% vs 37+/-2.6% at R30, 57+/-5.0% or 56+/-4.7% vs 45+/-2.7% at R60, 64+/-3.3% or 56+/-4.2% vs 48+/-3.4% at R90,respectively P【0.05). Blood flow changes were also reflected by lowering of jejunal PrCO2 measurements after intraluminal anisodamine or dobutamine compared with that of the saline controls (41+/-3.1 mmHg or 44+/-3.0 mmHg vs 49+/-3.7 mmHg at R30,38+/-3.7 mmHg or 40+/-2.1 mmHg vs 47+/-3.8 mmHg at R60,34+/-2.1 mmHg or 39+/-3.0 mmHg vs 46+/-3.4 mmHg at R90, respectively, P【0.05). Most interesting finding was that there were significantly higher mucosal blood flow and lower jejunal PrCO2 in anisodamine group than those in dobutamine group at 90 minutes of reperfusion(64+/-3.3% vs 56+/-4.2% for blood flow or 34+/-2.1 mmHg vs 39+/-3.0 mmHg for PrCO2, respectively, P【0.05), suggesting that anisodamine had a more lasting effect on mucosal perfusion than dobutamine. CONCLUSION: Intraluminal anisodamine and dobutamine can augment mucosal blood flow during gut I/R and alleviate mucosal acidosis. The results provided beneficial effects on the treatment of splanchnic hypoperfusion following traumatic or burn shock.

Ameliorating effect of Zhizi(Fructus Gardeniae) extract and its glycosides on scopolamine-induced memory impairment

OBJECTIVE： To study the chemical constituents of Zhizi （Fructus Gardeniae） and their antiamnesic ef- fect in a mouse model of Alzheimer＇s disease. METHODS： Ameliorating effects of the extracts, fractions and constituents on scopolamine-in- duced memory impairment in vivo using a passive avoidance task system and their inhibitory activi- ties on acetylcholinesterase （ACHE） in vitro were ex- amined. The isolation of components was per- formed by chromatographic techniques and their structures were identified on the basis of spectral analysis. RESULTS： Activity-guided fractionation of the total extracts resulted in the isolation of two glycosides, geniposide and cr0cin from the n-butanol fraction and genipin and crocetin from the ethylacetate fraction. Among the fractions tested, n-butanol fraction showed the strongest AChE inhibition （43.4% at a final dose of 0.03 mg/mL） and also ex- hibited outstanding efficacy （65.9% at a dose of2.50 mg/kg） in an experimental model of amnesia. Geniposide showed a 22.8% AChE inhibitory activi- ty and a potent ameliorating effect on scopol- amine-induced memory impairment in amnesic mice of 93.4% as compared to the control group. CONCLUSION： Geniposide, a main constituent of gardenia should be considered a candidate for fur- ther clinical study for the purpose of developing a cognition activator and its mechanism of action may be mediated, at least in part, by the acetylcho- line enhancing cholinergic nervous system.

THE EFFECT OF ANISODAMINE ON CEREBRAL RESUSCITATION OF RATS IN ACUTE CEREBRAL ISCHEMIA FROM CARDIAC ARREST

In order to investigate the mechanisms of acute cerebral ischemia, and to look for effective drugs on cerebral resuscitation, we made a model of acute complete global brain ischemia, reperfusion and resuscitation on rats according to Garavilla's method.Our results showed that the event of cerebral ischemia and reperfusion injury could resuh in the increase of total brain calcium content, and anisodamine has the same reducing brain calcium contents as diltiazem's, while improving neurological outcome and alleviating injury to neurons.

Unexpected anterograde amnesia associated with Buscopan used as a predmedication for endocscopy

It has been known that peripheral adverse event is caused by peripheral antimuscarinic action, from hyoscine butylbromide (Buscopan; Boehringer Ingelheim, Germany) used as a premedication for endoscopy. However, symptoms or signs associated with the central nervous system are rarely reported in the field of anesthesiology and peripartum labor. This central anticholinergic syndrome is likely caused by blockade of muscarinic cholinergic receptors in the central nervous system. There is no report on Buscopan-induced central anticholinergic syndrome in endoscopy room so far. Three middle-aged females unexpectedly suffered from anterograde amnesia after intramuscular injection of hyoscine butylbromide as an antispasmodic premedication for endoscopy at our endoscopy unit in the Health Promotion Center.

Effect of dexamethasone,anisodamine and rhubarb therapy on rats with acute pancreatitis

Objective:To investigate the therapeutic effects of dexamethasone,anisodamine and rhubarb(DAR) on endotoxin,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and pancreatic damage in rat models of acute pancreatitis(AP).Methods:The AP rat models were prepared and randomly assigned to AP group(n=10) and DAR group(n=10),while other healthy rats were assigned to the sham-operated group(n=10).The rats were euthanized at 6 h after operation,and then the serum levels of endotoxin,TNF-α,IL-6 and histology of pancreas were determined as the indexes of therapeutic effects.Results:At 6 h after operation,serum levels of endotoxin,TNF-α and IL-6,and pancreatic damage were significantly increased in AP group compared with those in sham-operated group(P<0.01).Compared with the AP group,DAR therapy remarkably attenuated the endotoxin,TNF-α,IL-6 levels and reduced pancreatic damage(P<0.05).Conclusion:The inhibition of pancreatic damage by DAR in rats with AP might contribute,in part at least,to the amelioration of pancreatic inflammation.The present study provides beneficial evidence that DAR may be useful in the treatment of AP model of rats.

Low dose transdermal scopolamine increases cardiac vagal tone in patients after acute myocardial infarction

OBJECTIVE: To investigate whether transdermal scopolamine increased cardiac vagal activity in patients during the acute phase of myocardial infarction. METHODS: 30 patients with a first acute myocardial infarction and preserved sinus rhythm who were on no drug that could influence the sinus node were randomly assigned to either treatment group or placebo group. Measures of heart rate variability (HRV) in patients given drug or placebo were obtained by digital 24 hour Holter recording before and after treatment. Baroreflex sensitivity was performed using the phenylephrine method. RESULTS: No significant differences was found in the indices of the time domain and the frequency domain in both groups before treatment. Patients with transdermal scopolamine showed a significant increase in the standard deviation of normal RR intervals (SDNN), standard deviation of all five min mean normal RR intervals (SDANN), root mean square of differences of successive normal RR intervals (rMSSD), total power (TP, 0.000. - 0.40 Hz), low frequency peak (LF, 0.040 - 0.15 Hz), high frequency peak (HF, 0.15 - 0.40 Hz), and Baroreflex sensitivity after treatment (P

Acupuncture combined with herb decoction for intractable hiccup

Objective To observe the efficacy of scalp acupuncture combined with oral administration of ènì Tāng（呃逆汤, Hiccup Decoction） for treatment of intractable hiccup. Methods One hundred patients with intractable hiccup were divided into treatment group（acupuncture combined with herb decoction group） and control group（western medicine group） according to the random number table method with 50 patients in each group. In treatment group, scalp acupuncture was conducted in the gastric area and thoracic area of patients, and Hiccup Decoction was taken orally. In control group, anisodamine（654-2） was injected intramuscularly in the dosage of l0 mg. The treatment was conducted for once a day in each group, and treatment for three consecutive times was needed. The number of times and the frequency of hiccup were observed after each treatment, and the results were compared. The standard of therapeutic effectiveness was evaluated by the number of hiccups decreased. Results In treatment group, the total cure rate was 54%, and the total effective rate was 84%, in control group, the total cure rate was 32%, and the total effective rate was 66%. According to the comparison between the two groups in the cure rate and total effective rate, the differences were statistically significant（both P〈0.05）. After the first treatment, there was no statistical difference according to the comparison between the two groups in the cure rate and total effective rate（both P〈0.05）, after the second treatment, the differences were statistically significant according to the comparison between the two groups in the cure rate and total effective rate（both P〈0.05）, and the efficacy in treatment group was obviously superior to that in control group, after the third treatment, there was no statistical difference according to the comparison between the two groups in the cure rate and total effective rate（both P〈0.05）. Conclusion In the one course of treatment, the efficacy of acupuncture combined with herb decoction for treatment of intractable hiccup was obviously superior to that of intramuscular injection with anisodamine. The therapeutic effect of intramuscular injection with anisodamine for treatment of intractable hiccup was the best at the first treatment, and the therapeutic effect of acupuncture combined with herb decoction was the best at the second treatment.

Protective effect of anisodamine on respiratory function after severe brain inju ry

To study the clinical therapeutic e ffect of anisodamine on respiratory function after severe brain injury. Methods: Ninety patients with respiratory dysfunction following severe brain injury were divided into two groups: a treatment group (n=45, trea ted with routine therapy plus anisodamine) and a control group (n=45, treated wi th routine therapy only). The pulmonary ventilation function and oxygenation fun ction were compared between the two groups. Results: In the treatment group, 12 hours after treatment the r espiratory rate reduced, the partial pressure of carbon dioxide (PCO 2), the pa rtial pressure of oxygen in arterial blood (PaO 2) and oxygenation exponent inc reased, the dead space ventilation dose and the pulmonary alveolus partial pres sure of arterial oxygen difference decreased, and the ventilation function of th e respiratory tract and pulmonary oxygenation function improved. There was a sig nificant difference between the two groups (P< 0.01 ). N o side effect was found except a slight increase of intracranial pressure and h eart rate. Conclusions: Anisodamine can improve pulmonary ventilation func tion and oxygenation function and decrease the incidence of hypoxemia markedly. It is effective in treating respiratory dysfunction after severe brain injury.

Ameliorating effects of constituents from Cortex Acanthopanacis Radicis on memory impairment in mice induced by scopolamine

OBJECTIVE:To investigate the effects of active constituents extracted from Cortex Acanthopanacis Radicison improving the impaired memory in mice models.METHODS:The mice models of memory impairment were established using scopolamine.Ameliorating effects of the fractions and constituents on scopolamine-induced memory impairment in vivo were investigated using passive avoidance and Morris water-maze task tests,and their anti-acetylcholinesterase(AChE)and antioxidant activities in vitro examined.The isolation of constituents was performed by chromatographic methods and their structures were identified on the basis of instrumental analysis.RESULTS:Among the fractions tested,ethylacetate fraction exhibited the anti-AChE activity(25.83%±0.23%)properly and excellent 2,2-diphenyl-1-picrylhydrazyl(DPPH)radical and superoxide anion scavenging capacity(87.50%±0.83%and 60.22%±0.43%,respectively).However,the methylene chloride fraction was much more active than the ethylacetate fraction in the passive avoidance task test(167.5%increase of step-through latency time)and Morris water-maze task test(33.3%decrease of es-cape latency time).Four constituents,β-sitosterol,stigmasterol,sesamin,and hyperin were isolated from the methylene chloride fraction,among them,hyperin showed anti-acetylcholinesterase and anti-oxidant activities remarkably.Moreover,hyperin exerted a potent effect(146±38)s on memory improvement in terms of passive avoidance task test compared with the reference compound tacrine(162±43)s at a dose of 2.5 mg/kg.CONCLUSION:Hyperin,a flavonoid glucoside isolated from Cortex Acanthopanacis Radicis,inhibited AChE activity and potently ameliorated scopolamine-induced memory impairment,and its action may be partially mediated by the acetylcholine-enhancing cholinergic nervous system.