广西眼镜王蛇毒酸性磷脂酶A2的分离纯化和性质研究

目的广西眼镜王蛇毒酸性磷脂酶A2的分离纯化和性质研究.方法分离纯化采用SephadexG-75,CM-SepharoseCL-6B和DEAE-SephadexA-50柱层析法;通过电泳,磷脂酶A2活力测定,氨基酸组成测定,N端序列分析及药理实验进行性质研究.结果从广西眼镜王蛇毒中分离到一种酸性的磷脂酶A2(命名为APLA2-1),相对分子质量为14600u,等电点5.4,酶的比活力为108μmol/mg*min-1,N端序列为NLIQFGNMIQCTVPGF.它对实验用小白鼠有致死性(LD506.6mg/kg),具有肌毒性,心脏毒性,抗血小板聚集活性,能诱导水肿形成.结论APLA2-1生化和药理性质的研究,为阐明蛇毒PLA2结构和功能关系,及蛇伤中毒机制,打下了良好基础.     

广西眼镜蛇毒中Natrin蛋白的分离及纯化

目的:从广西眼镜蛇毒中分离出Natrin蛋白.方法:广西眼镜蛇毒经SephadexG75凝胶、CM Sepharose CL-6B离子交换层析及FPLC mono-S柱层析分离纯化出Natrin蛋白.结果:从广西眼镜蛇蛇毒中分离纯化得到的Natrin蛋白,经SDS-PAGE电泳测定其分子量为25 ku,质谱测定其分子量...     

自制CM-C对眼镜王蛇蛇毒的柱层析及其中五种酶的分布

眼镜王蛇(Ophiophagus hannah)广泛分布于我国北回归线以南各省区,我区以河池、柳州、百色、南宁和玉林等专区为多.Joubert用柱层析分离出眼镜王蛇蛇毒两种神经毒素,并测定其氨基酸序列.Mebs测定了眼镜王蛇原蛇毒某些酶的活性.1979年我院     

福建产眼镜王蛇毒神经毒素的分离及活性测定

目的 从眼镜王蛇蛇毒中分离神经毒素，测定其活性和毒性。方法 应用CM—Sephadex C-50离子交换色谱，阶段梯度洗脱分离粗毒；用Sephadex G-50凝胶过滤纯化组分；用大鼠体外膈神经-膈肌标本鉴定分离组分对神经-肌肉接头的作用性质；观察神经毒素对乙酰胆碱(Ach)引起蛙体外腹直肌收缩的量效曲线的影响；以Meier法测定神经毒索对小鼠的半数致死量(LD50)。结果 眼镜王蛇粗毒经阳离子交换色谱后获得A～N共14个蛋白峰，经鉴定E、F、G，H、I、K等6个蛋白峰具有阻断神经一肌肉接头传导的作用，被鉴定为神经毒素，约占粗毒的48．3％；通过凝胶过滤对E、G和K组分进行初步纯化；E、G和K组分可使Ach引起的蛙腹直肌收缩的量效曲线平行右移，它们与N2-胆碱受体(N2-AchR)的解离常数(KD)分别为：145．87，73．53和20．04ng／mL；E、G和K组分小鼠静脉注射的LD50分别为：0．884，0．749和1．58mg／kg。结论 眼镜王蛇粗毒经离子交换色谱获得6个神经毒索组分，经鉴定E、G和K组分为N2-AchR的竞争性拮抗剂。     

广西眼镜王蛇蛇毒磷脂酶A_2的分离纯化及电镜观察

广西眼镜王蛇(Ophiophagus hannah)蛇毒,经羧甲基纤维素离子交换层析,可得20个蛋白峰,磷脂酶A_2活性以第一峰活性最强,第二和第十二峰也有部分活性.取第一峰观察其对红细胞电泳速度的影响及对红细胞膜影响的电镜观察,发现磷脂酶A_2对红细胞电泳速度有明显的延缓作用,并对人的红细胞膜有明显的作用,但对山羊红细胞膜未见明显改变.研究还表明眼镜王蛇毒中磷脂酶A_2也具有很好的热稳定性.     

广西眼镜蛇毒细胞毒素对人卵巢癌细胞和宫颈癌细胞的抑制作用研究

目的：研究广西眼镜蛇毒细胞毒不对体外培养的人卵巢癌细胞和宫颈癌细胞株的抑制作用。方法：通过SephadexG-50,GM-Sepharose,CL-6B柱层析从广西眼镜蛇毒中分离纯化细胞毒素,应用四素氮唑蓝（MTT）法测定其对人卵巢癌细胞和宫颈癌细胞株的体外细胞毒作用及其最效关系,并与粗毒的抑瘤作用进行比较,结果：从广西眼镜蛇毒中分离到一种细胞毒素（CTX CM－5）,其对体外培养的人卵巢癌细胞和宫颈癌细胞株的抑制作用呈明显的剂量依赖关系。抑制率随剂是增加而呈迅速上升培养的人卵巢 细胞和宫颈癌细胞株具有较强的抑制作用。     

福建产舟山眼镜蛇毒细胞毒素的快速分离纯化及鉴定

目的 从舟山眼镜蛇毒中分离纯化细胞毒素(CTX)，并鉴定其理化性质。方法 采用SP—Sephadex C-25阳离子交换色谱及Sephasil Peptide C18反相高效液相色谱法从舟山眼镜蛇毒中分离纯化CTX，SDS—PAGE(Tris—Tricine系统)鉴定纯度，Edman降解法测定N端氨基酸序列。结果 粗毒经阳离子交换色谱，得到14个蛋白峰，其中第X～XⅢ峰具CTX活性；再分别经反相色谱纯化，得到4个CTX．总得率为32.48％；SDS—PAGE显示为均一蛋白，分子量依次为：7．28，7．33，7．24和7．38kD；测定它们N端20个氨基酸序列。结论 采用阳离子交换和反相高效液相色谱可快速、高效地从眼镜蛇毒中获得4个CTX纯品。     

舟山眼镜蛇毒L-氨基酸氧化酶的分离纯化、理化及酶学性质

目的 从舟山眼镜蛇蛇毒中分离L-氨基酸氧化酶(LAO),测定其理化和酶学性质.方法 应用Sephadex G-100凝胶色谱和POROS CM20离子交换色谱分离纯化LAO;Wellner和Lichtenberg法测定LAO活力,SDS-PAGE法测定相对分子质量.结果 舟山眼镜蛇蛇毒经Sephadex G-100凝胶色谱和两次POROS 20离子交换色谱后,获得LAO纯品(暂定名NA-LAO).NA-LAO在非还原和还原条件下相对分子质量均为58 kD左右;其反应最适pH为8.0,最适温度为60℃,对L-苯丙氨酸的米氏常数(Km)为3.08 mmoL/L.结论 应用凝胶过滤色谱和离子交换色谱可从舟山眼镜蛇毒中分离纯化LAO.     

眼镜王蛇神经毒素的提纯及其性质的研究

本文用SP-SephadexC-25柱层析法分离广西眼镜王蛇蛇毒,得到19个组分,其中7个组分有毒性。对毒性较强,含量较高的4个毒性组分(即神经毒素Ⅴ、Ⅶ、Ⅷ和Ⅹ)用SephadexG-50凝胶过滤法进行纯化,并对神蛇经毒素Ⅶ和Ⅹ用CM-SephadexC-50柱层析法做了进一步纯化。经聚丙烯酰胺凝胶电泳(PAGE)鉴定,这4个神经毒素均为单一条带。用SDS-PAGE法测得神经毒素Ⅴ、Ⅶ、Ⅷ和Ⅹ的分子量分别为7800,6700,8400和8000,并用它们进行了小鸡颈二腹肌神经—肌肉接头传递的阻遏试验,结果表明这4个神经毒素均有阻遏神经—肌肉接头传递的作用,而且阻遇作用基本不可逆,属于突触后神经毒素。     

蝮蛇蛇毒类纤溶酶的分离纯化及一些特性的研究

蝮蛇(Agkistrodon haly palls)蛇毒先用Sephadex G-100柱层析分离出有纤溶活性的第2、3峰,再用DEAE-Sephadex A-50分离有纤溶活性的第14峰,经PAA圆盘电泳仍有多条区带.该组分用高效液相配用Mono-Q阴离子交换柱层析二次纯化,得有纤溶活性的第2峰,PAA圆盘电泳为第二条紧靠的区带,SDS/PAGE测得该纤溶组分分子量为21000±1000;pI为3.9±0.05,对酪蛋白有一定水解能力,EDTA能降低酶活性,氨基酸组成分析为酸性氨基酸居多的蛋白质.     

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.