微量元素对黄芩有效成分积累的影响

目的研究微量元素锌、镁对黄芩组织生长情况和有效成分积累的影响。方法改变MS培养基中微量元素锌和镁的含量对黄芩愈伤组织进行继代培养;用UV-Vis法和HPLC法分别测定了继代培养所得愈伤组织中提取的总黄酮和黄芩苷的含量。结果 Zn2+浓度0.045 mmol/L、Mg2+浓度2.25 mmol/L情况下有利于黄芩愈伤组织生长;增大Zn2+浓度(0.045 mmol/L)并降低Mg2+浓度有利于黄芩愈伤组织中黄酮类物质的合成;低浓度的Mg2+有利于黄芩愈伤组织中黄芩苷的生物合成。结论通过调控微量元素锌和镁的量可以有效提高黄芩愈伤组织生长率并显著提高其有效成分的含量。     

两种金属离子对黄芩愈伤组织培养及其次生代谢物积累的影响

目的研究Fe%^2＋、Mn^2＋离子对黄芩组织培养过程中组织生长量和次生代谢物积累的影响。方法进行黄芩愈伤组织的诱导、分化，然后改变MS培养基中Fe^2＋、Mn^2＋的含量进行继代培养；提取所得黄芩愈伤组织中的黄酮类化合物，分别用紫外可见分光光度法和高效液相色谱法测定了总黄酮和黄芩苷的含量。结果有利于黄芩愈伤组织生长的金属离子浓度为：Fe^2＋0.15mmol/L、Mn^2＋0.2mmol/L；增大Mn^2＋浓度到1.5mmol/L有利于黄酮类次生代谢物生物合成；而高浓度的Fe^2＋有利于黄芩苷的生物合成。结论通过调节金属离子含量可以有效的提高黄芩愈伤组织生长率并显著提高其有效成分的含量。     

黄芩愈伤组织中多酚氧化酶活性与黄芩苷合成的关系研究

目的 研究黄芩愈伤组织中多酚氧化酶活性变化规律及其与黄芩苷次生合成之间的关系。 方法 采用紫外分光光度法测定多酚氧化酶活性,采用高效液相色谱法测定黄芩苷的量。研究不同添加物(抗坏血酸、氯化钠、苯甲酸、聚乙烯吡咯烷酮、硫酸铜)对多酚氧化酶和黄芩苷量的影响。 结果 在黄芩愈伤组织生长周期的前20 ｄ,黄芩苷基本没有合成,多酚氧化酶的活性低水平表达;20～35 ｄ,黄芩苷大量合成,多酚氧化酶的活性有所降低,但降低趋势较小;35～50 ｄ,多酚氧化酶活性高水平表达,黄芩苷次生合成受到抑制。 结论 多酚氧化酶的高水平表达不利于黄芩苷的次生合成,抗坏血酸、氯化钠、苯甲酸等均可抑制多酚氧化酶的活性,促进黄芩苷次生合成,其中以添加0.02%的抗坏血酸效果最为明显,可使黄芩苷的量提高17.6%(82.3 ｍｇ／ｇ),而聚乙烯吡咯烷酮、硫酸铜对多酚氧化酶的活性有着明显的促进作用,抑制了黄芩苷的积累。     

黄芩不同器官及愈伤组织中3种黄酮类成分和总黄酮的含量比较

目的?比较黄芩植株不同器官（根、茎、叶、花）及黄芩药材（子芩、枯芩、绿芩）和黄芩愈伤组织中野黄芩苷、黄芩苷、汉黄芩苷和总黄酮的含量。方法?采用SYMMETRY C₁₈（4.6?mm×250?mm,5?μm）柱,以乙腈（A）-0.22%乙酸水溶液（B）为流动相,柱温35?℃,波长335?nm,检测3种黄酮类成分的含量;采用紫外分光光度法,以芦丁为对照品,采用硝酸铝显色后于504?nm处测定其总黄酮含量。结果?野黄芩苷、黄芩苷、汉黄芩苷在质量浓度为6.0~192.0、30.0~960.0、12.0~384.0?μg/mL范围内与峰面积呈良好线性关系（R2≤0.998?9）,平均加样回收率为97.8%~103.1%,RSD≤4.0%。总黄酮在质量浓度为40.00~240.00?μg/mL范围内线性关系良好（R2≥0.999?8）,平均加样回收率为98.8%,RSD为1.0%。结论?黄芩地上部分（花、茎、叶）的野黄芩苷含量高于地下部分（根）,以花中的含量为最高,达到0.53%;地下部分的黄芩苷、汉黄芩苷含量高于地上部分,愈伤组织中黄芩苷达到0.60%。总黄酮含量以愈伤组织中含量最高,达到30.42%。为利用黄芩废弃部位花提取野黄芩苷和利用愈伤组织生产黄酮提供了依据。      

诱导子对黄芩毛状根生长及黄芩苷合成的影响

目的 考察诱导子对黄芩毛状根生长和黄芩苷生物合成的影响。方法 用非生物诱导子(Ca²⁺、Mg²⁺和Co²⁺)和从黑曲霉Aspergillus niger、米曲霉A. oryzae、蜜环菌Armillaria mellea中提取的真菌诱导子,与黄芩毛状根共培养;HPLC法测黄芩苷的量。结果 各诱导子对黄芩毛状根生长和黄芩苷合成有不同影响：黑曲霉诱导子和米曲霉诱导子质量浓度为40和20 mg/L时,Co²⁺浓度为1.53×10^-4 mmol/L时,黄芩苷的量从7.64%分别增至9.18%、8.81%和8.62%。结论 诱导子对黄芩毛状根次生代谢产物具有特异性,可选择性地促进某一类次生代谢产物的合成。试验证实黑曲霉诱导子、米曲霉诱导子和Co²⁺是适合黄芩毛状根代谢黄芩苷调控的诱导子。     

甘草酸对黄芩苷增溶作用的研究

目的 通过测定不同配比黄芩甘草合煎液的表面张力及指标性成分含量,探讨甘草酸对黄芩苷增溶作用的机制.方法 采用Wilhelmy吊片法测定不同配比黄芩甘草合煎液的表面张力;采用HPLC测定不同配比条件下黄芩苷含量和甘草酸的浓度,并作出二者之间的关系图.结果 黄芩甘草的比例为3∶2时,黄芩苷的提取率最高,此时甘草酸的临界胶束浓度是0.22 g/L.在0.22 g/L之前,随着甘草酸浓度的增大,黄芩苷的溶出度逐渐增大;当甘草酸浓度达到0.22 g/L后,黄芩苷的溶出度不再增加.结论 甘草酸对黄芩苷有一定的增溶作用,并且在达到它的临界胶束浓度0.22 g/L时,增溶作用最大.     

黄芩主要成分体外抗甲型流感病毒作用的研究

目的比较黄芩的主要成分黄芩苷、黄芩素、汉黄芩素体外抗甲型流感病毒FM1感染的作用,明确黄芩抗流感病毒的主要有效成分。方法采用四甲基偶氮唑蓝(MTT)比色法检测黄芩苷、黄芩素、汉黄芩素对狗肾细胞株(MDCK)的最大无毒浓度(TC0)及半数中毒浓度(TC50),通过细胞病变效应(CPE)法观察药物对病毒致细胞病变作用的影响。MTT法检测药物不同浓度的抗病毒有效率,Probit回归法计算黄芩苷、黄芩素、汉黄芩素对病毒感染的半数抑制浓度(IC50)及治疗指数(TI),荧光法检测黄芩苷对病毒神经氨酸酶活力的影响。结果黄芩苷对流感病毒所致的细胞病变作用有明显的抑制作用,当黄芩苷浓度在0.0156～0.1250g/L时,细胞存活率明显高于病毒感染组,其中以0.1250g/L组的细胞存活率最高,抗病毒有效率达94.6%,其IC50为0.0162g/L,TI为21.34;黄芩素对流感病毒所致的细胞病变有一定的抑制作用,当黄芩素浓度在0.0860g/L时,抗病毒有效率达48.72%,其IC50为0.0778g/L,TI为8.89;汉黄芩素对流感病毒所致的细胞病变无抑制作用,其IC50为0.0236g/L,TI为0.051。当黄芩苷浓度?0.0313g/L时,能明显降低病毒神经氨酸酶的活力。结论黄芩主要成分中抗流感病毒的作用为黄芩苷黄芩素汉黄芩素,说明黄芩抗流感病毒的主要有效成分是黄芩苷。     

黄芩苷对高糖环境下肾小球系膜细胞增殖的影响

目的 探讨不同剂量的黄芩苷对高糖环境下肾小球系膜细胞(GMC)增殖的影响,以期探索黄芩苷防治DN的药效机制,为中医药防治DN提供新的视角和实验根据.方法 体外培养大鼠肾小球系统细胞(GMC)并分为9组:正常组(LG 5.5mmol/L)、模型组(HG 25mmol/L)、黄芩苷1组(HG 25mmol/L+黄芩苷3.125μmol/L)、黄芩苷2组(HG 25mmol/L+黄芩苷6.25μmol/L)、黄芩苷3组(黄芩苷(HG 25mmol/L+黄芩苷12.5μmol/L)、黄芩苷4组(HG25mmol/L+黄芩苷100μmol/L)、黄芩苷5组(HG 25mmol/L+黄芩苷50μmol/L)、黄芩苷6组(HG 25mmol/L+黄芩苷100μmol/L)和PKC抑制剂组(HG 25mmol/L+ PKC抑制0.5 nmol/L),每组设平行6孔,培养24h、48h、72h后,应用MTT比色法检测各组GMC增殖情况,倒置显微镜下观察各组GMC生长情况,评价黄芩苷对GMC生长的影响.结果 从各时间点测得的OD值可见,用药组与高糖组比较差异有统计学意义(P<0.05),黄芩苷组与PKC阳性药组比较差异均有统计学意义(P<0.05).从抑制率可见,24h时,除了黄芩苷6组(即给药剂量为100μmol/L)对高糖刺激后GMC增殖有抑制作用之外,其余各组对高糖刺激后GMC增殖均无抑制.48h、72h随着时间增加,各组对高糖刺激后GMC增殖的抑制率逐渐升高,且随着黄芩苷浓度增加抑制率逐渐增高.结论 高糖环境下,体外培养GMC存在异常增殖现象,黄芩苷对高糖刺激GMC增殖有抑制作用,且具有一定剂量效应关系.即抑制率随着黄芩苷浓度的增高而升高,尤其是25μmol/L、50μmol/L、100μmol/L三组的时间剂量依赖关系更加明显,且在72h的时间点抑制率最高,抑制效果最佳.     

南方红豆杉愈伤组织生长动力学及紫杉醇代谢动力学研究

目的 通过愈伤组织细胞生长及其紫杉醇代谢动力学的研究,筛选确定南方红豆杉愈伤组织培养生产紫杉醇的最佳培养基配方与最佳收获期。方法 以改良MS为基本培养基添加不同质量浓度的IBA、6-BA、GA₃组合诱导培养南方红豆杉愈伤组织,通过测定愈伤组织鲜质量进行生长动力学研究,采用HPLC法检测不同生长阶段愈伤组织中紫杉醇的积累量,进行紫杉醇代谢动力学研究。结果 改良MS＋0.2 mg/L IBA＋0.05 mg/L 6-BA＋0.3 mg/L GA₃、MS＋0.2 mg/L IBA＋0.01 mg/L 6-BA＋0.7 mg/L GA₃及MS＋0.2 mg/L IBA＋0.1 mg/L 6-BA＋0.5 mg/L GA₃培养条件均较有利于南方红豆杉愈伤组织的诱导培养,且紫杉醇的积累量也较高,最高可达0.037 76%。结论 在添加IBA及6-BA的基础上适当增补GA₃可使南方红豆杉愈伤组织诱导提前启动,缩短出愈时间,促进生长,降低褐变程度,增加愈伤组织中紫杉醇的积累量。愈伤组织的生长曲线呈S型,紫杉醇的积累呈线型增加,是一个逐渐积累的过程,但积累到一定程度就不再增加,且伴随着愈伤组织不断褐变。     

不同浓度外源激素对黄芩愈伤组织诱导影响

目的：运用植物组织培养技术选择适合黄芩愈伤组织诱导的生长素与细胞分裂素的配比及浓度。方法：以黄芩幼根、叶片和带节茎段为外植体,用不同浓度生长素和细胞分裂素对黄芩进行愈伤组织诱导试验。结果：6-BA1.0mg·L-1诱导形成的愈伤组织疏松,诱导率高。诱导黄芩愈伤组织最佳生长素与细胞分裂素的浓度配比为NAA0.5mg·L-1＋6-BA1.0mg·L-1。结论：生长素与细胞分裂素配合使用时,愈伤组织的诱导率明显高于单个激素的处理,诱导结果也有差异。     

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.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Electro-Acupuncture Combined with the Trigger Point Needle-Embedding for Treatment of Primary Trigeminal Neuralgia in 31 Cases

In recent years, the author of this essay has applied electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases, yielding satis- factory results as reported in the following.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Persist in and Carry Forward the Integrative Medical Research——Speech at the 6th National General Congress of Chinese Association of Integrative Medicine by Academician CHEN Zhu,Minister of Ministry of Health

The 6th National General Congress of Chinese Association of Integrative Medicine （CALM） was convened at 19-20, April 2008 in Beijing. Academician CHEN Zhu, the minister of Ministry of Health indicated at the congress that the integration of Chinese and Western medicine is very well in keeping with the situation of our country and the general rule of development in medical science; and as a good integration of Chinese medicine and Western medicine, it is mutually beneficial and advantageous to both of them. Seeing the creativity shown in integrative medical investigation in theoretic and methodological sides, we should and must persist in and develop it.