蟾酥抗肿瘤有效成分的活性追踪分离及急性毒性研究

目的 从传统中药蟾酥中寻找抗肿瘤活性强而毒性低的化合物。方法 采用追踪分离的方式,以体外肿瘤细胞生长抑制强度为活性指标,同时结合小鼠急性毒性实验,对蟾酥的主要化学成分进行初步研究。结果 采用活性追踪分离方式,从蟾酥中分离得到具有抑制肿瘤细胞生长活性同时对小鼠急性毒性较低的化合物蟾毒它灵,其对人非小细胞肺癌A549细胞具有较好的生长抑制作用,小鼠iv给药的LD₅₀约为蟾毒灵的4倍,而且稳定性较好。结论 综合化合物的活性、毒性及稳定性等因素,初步判断蟾毒它灵较其他同类化合物具有更好的抗肿瘤药物开发前景。     

应用HPLC法测定六应丸中蟾毒内酯的含量

应用ＨＰＬＣ法测定六应丸中蟾毒内酯的含量黄东亮黄小玲１（深圳制药厂，广东５１８０２９；１深圳市慢性病防治院）关键词六应丸；蟾毒内酯；高效液相色谱法中图号Ｒ９２７．２蟾酥是蟾蜍科动物中华大蟾蜍Ｂｕｆｕｇａｒｇａｒｉｚａｎｓｃａｎｔｏｒ或黑眶蟾蜍Ｂｕｆｕ...     

微乳毛细管电泳法测定蟾酥及其制剂中蟾毒内酯类成分的含量

目的应用微乳毛细管电泳法测定蟾酥及其制剂中蟾毒内酯类成分的含量。方法运行微乳缓冲液由0.81%正庚烷、6.61%正丁醇、3.31%SDS及89.27mmol/L硼砂溶液组成,pH=9.2。运行电压15kV(恒压分析);柱温40℃;检测波长298nm。结果华蟾毒精和酯蟾毒配基的线性范围分别为54.8~548.0mg/L及51.6~516.0mg/L;平均回收率分别为97.29%~99.82%(RSD为1.86%~2.91%,n=6)及97.52%~101.69%(RSD为2.14%~3.02%,n=6)。结论该方法简便、准确、重现性好,可作为蟾酥及其制剂中蟾毒内酯类成分的质量控制方法。     

蟾毒灵、华蟾酥毒基及酯蟾毒配基大鼠体内药动学研究

目的 建立检测大鼠血浆中3种蟾蜍甾烯类化合物蟾毒灵、华蟾酥毒基及酯蟾毒配基的HPLC法,并用于蟾酥在大鼠体内的药动学研究。方法 分别于大鼠尾iv给予蟾酥提取物0.8 mg/kg后2、5、10、15、20、30、45、60、90 min,眼眶取血,采用乙腈沉淀蛋白与液液萃取相结合方法进行血浆样品预处理,以HPLC法测定大鼠血浆中蟾毒灵、华蟾酥毒基及酯蟾毒配基的质量浓度,以Kinetica软件拟合药动学参数。结果 蟾毒灵、华蟾酥毒基及酯蟾毒配基均得到很好的分离,重现性、精密度、线性关系良好,达到体内分析要求;经非房室模型拟合,得到蟾毒灵、华蟾酥毒基及酯蟾毒配基在大鼠体内主要药动学参数;iv给药30 min时,血药浓度均降至C_max的1/5以下。结论 建立的蟾毒灵、华蟾酥毒基及酯蟾毒配基血药浓度测定方法操作简单、结果准确可靠;蟾蜍甾烯类成分在大鼠体内代谢较为迅速,所得数据可为蟾酥提取物的药动学研究提供参考。     

脂蟾毒配基、华蟾酥毒基、蟾毒灵配伍对BEL-7402人肝癌细胞的抑制作用

目的:研究蟾酥中脂蟾毒配基、华蟾酥毒基和蟾毒灵三种成分单体及配伍对BEL-7402人肝癌细胞的抑制作用。方法:采用MTT法测定蟾毒灵、华蟾酥毒基、脂蟾毒配基单体不同浓度和时间点,三种成分两两配伍以及三种成分配伍对BEL-7402人肝癌细胞增殖的抑制作用。结果:蟾毒灵对BEL-7402人肝癌细胞增殖的抑制作用最强,其次是华蟾酥毒基,脂蟾毒配基的作用最弱;脂蟾毒配基不能增强华蟾酥毒基、蟾毒灵的作用,且有抑制作用的趋势;华蟾酥毒基与蟾毒灵混合使用与单独使用的抑制作用无明显区别。结论:欲发挥三种成分的抗肿瘤作用,应该单独使用,或者华蟾酥毒基与蟾毒灵混合使用。     

脂蟾毒配基、华蟾酥毒基、蟾毒灵不同配伍的镇痛作用比较

目的:研究蟾酥中脂蟾毒配基、华蟾酥毒基和蟾毒灵三种成分配伍后的镇痛效果,以筛选出三种成分的最佳配伍方式。方法:采用醋酸致小鼠扭体法进行镇痛实验研究。结果:蟾毒灵的镇痛作用最强,其次是华蟾酥毒基,脂蟾毒配基的作用最弱;脂蟾毒配基用量较大时,与华蟾酥毒基配伍表现为相互的抑制作用,用量较小时则为协同作用;脂蟾毒配基与蟾毒灵配伍表现为相互的抑制作用;华蟾酥毒基与蟾毒灵配伍主要表现为蟾毒灵的镇痛作用;三者共同配伍时,未表现出相互间的协同或抑制作用。结论:欲发挥三种成分的镇痛作用,应该单独使用,或者华蟾酥毒基与蟾毒灵混合使用,以保证其安全性、有效性和质量可控性。     

三种蟾毒单体对SMMC-7721和BEL-7402人肝癌细胞生长的抑制作用

目的：探讨蟾蜍中主要蟾毒内酯类成分蟾毒灵、华蟾酥毒基、酯蟾毒配基的抗肝癌作用及机制。方法：以SMMC—7721、BEL—7402人肝癌细胞为靶细胞，以丝裂霉素为阳性对照药物，应用四甲基偶氟唑盐比色法、锥虫蓝染色和DNA染色分析法观察这3种蟾毒单体对肝癌细胞的生长抑制和诱导凋亡作用。结果：蟾毒灵和华蟾酥毒基对上述肝癌细胞具有明显生长抑制作用，华蟾酥毒基较蟾毒灵作用略弱，但两者达到相同生长抑制效果所需浓度均低于丝裂霉素，酯蟾毒配基只有在较高浓度下才显示对肝癌细胞的生长抑制作用；不同浓度蟾毒灵、华蟾酥毒基对人肝癌细胞细胞膜具有破坏作用；可使人肝癌细胞阻止于G2／M期，使处于S期的细施比例降低，并有明显诱导肝癌细胞凋亡作用。结论：蟾毒灵、华蟾酥毒基对上述人肝癌细胞具有明显生长抑制作用，具有对肝癌细胞膜的直接破坏作用和诱导凋亡作用；酯蟾毒配基抗肿瘤作用较弱。     

HPLC法测定"心力丸"中华蟾酥毒基和脂蟾毒配基的含量

目的建立测定心力丸中华蟾酥毒基和脂蟾毒配基含量的方法。方法采用反相高效液相色谱法,色谱柱Spherisorb C18(5μm,4.6mm×200 mm),流动相为0.5%(ω)磷酸二氢钾溶液-乙腈(体积比60∶40)(用磷酸调节pH值为3.2),流速1.0 mL/m in,柱温35℃,检测波长为296 nm。结果华蟾酥毒基回收率99.20%,RSD=1.27%;脂蟾毒配基回收率99.40%,RSD=0.86%。结论本法简便、准确,可用于控制该制剂质量。     

脂蟾毒配基对失血性休克和实验性心衰的作用

脂蟾毒配基(resibufogenin, RBG)是从中药蟾酥中提得的具有较强生物活性的物质。其结构类似于洋地黄毒甙配基。现认为RBG是蟾酥甾族化合物中毒性最小的一种。其安     

华蟾素精和羟基华蟾毒精的人肠内细菌代谢研究

目的：研究人肠内细菌对蟾酥中主要成分华蟾毒精和羟基华蟾毒精的代谢。方法：采用体外人肠内细菌粗酶与华蟾毒精和羟基华蟾毒精温孵法;根据波谱学和化学数据鉴定代谢产物的结构;根据原形化合物和代谢产物的体外抑制人癌细胞系（HCT-8、KB、BGC、BIU和HeLa)的生长评价其抗肿瘤活性。结果：华蟾毒精可由人肠内细菌代谢产生去乙酰基华蟾毒精;羟基华蟾毒精可由人肠内细菌代谢产生去乙酰基羟基华蟾毒精;原形化合物具有强抑制人癌细胞生长的活性,而其代谢产物无活性。结论：华蟾毒精和羟基华蟾毒精由肠内细菌代谢而失活。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.     

Possible Spread of adenovirus type 3 from poultry to humans ：indirect evidence from an outbreak in China

Objective:To explore the epidemiology and etiology for an outbreak of acute respiratory tract infection that occurred in one county of Jiangsu Province, China 2004. Methods: Only cases meeting the case definition were included in the study. We reviewed the medical records of the cases who were admitted to the local hospitals, interviewed cases by a standard questionnaire, and then described the epidemiotogic features and analyzed risk factors by means of a case-control study. We collected pharyngeal swab specimens and sent them to different laboratories for isolation and culture. The laboratory used different detection methods such as DIP, PCR, electron microscope examination and microneutralization assay, to identify and then type the positive specimens. Results:A total of 871 cases were reported during the period from April 18 to July 4,2004. The distribution of onset times presented two peaks, one in late May and another in middle June. The epidemic occurred mainly in the elementary and junior high schools in ten townships of one county, and the mean age of the cases was 12 years (range 7 months to 18 years). The course of the disease was acute, and was characterized by fever accompanied with sore throat and tonsillitis. The WBC count of cases was normal or elevated. The mean duration of illness was 5 days (range 2 to 12 days). No fatalities from illness were reported. A case-control study indicated that the possible risk factors were close contact with a case and/or poultry before onset and sharing of towels among members of the family. The typical CPE was observed through inoculating pharyngeal swab specimens into the HEP-2 cell cultures in different laboratories. An infection of adenovirus type 3 was verified by detecting positive specimens in different methods. Conclusion:This investigation demonstrated that the acute respiratory infection in cases was caused by adenovirus type 3. Cases occurred in over 70 schools in ten townships in 2004, and the route of transmission was possibly close contact with cases or droplet transmission.