一株海洋芽孢杆菌的鉴定及抗菌活性研究

目的:对一株编号为ZJ8112的分离自东海海域海泥样品的细菌进行鉴定和抗菌活性研究.方法:采用传统的分类学方法观察菌株ZJ8112的形态、耐盐度,结合16S rDNA 序列分析在GenBank上进行比对以确立其进化地位.利用稻瘟霉菌(Pyricularia oryzae)P-2b、枯草芽孢杆菌(Bacillus subtilis)、大肠埃希菌(Escherichia coli)、啤酒酵母(Saccharomyces cerevisiae)、白念珠菌(Candida albicans)和黄瓜黑星病菌(Cladosporium cucumerinum)作为抗菌活性指示菌,检验ZJ8112的抗菌活性.结果:枯草芽孢杆菌ZJ8112革兰染色阳性,芽孢中生.其最佳生长盐度为10%,属于中等嗜盐菌.通过16S rDNA序列分析发现其与Bacillus subtilis F121112在进化位置上最为接近.该菌株的16S rDNA序列已经提交GenBank,接受号为EU100745.综合ZJ8112菌株的培养特性和形态特征,可认为ZJ8112是一株海洋芽孢杆菌.抗菌活性研究的结果表明其具有很强的抗稻瘟霉菌和抗大肠埃希菌活性.结论:ZJ8112为中等嗜盐的海洋细菌,其最适盐度为10%.生物学活性研究的结果表明,其发酵液具有很强的抗稻瘟霉活性和抗大肠埃希菌活性,具有潜在的抗生素研究的价值.     

两株产生大环内酯Macrolactin S的海洋细菌的筛选及鉴定

目的:从中国东海分离筛选抗菌活性微生物,确定活性菌株的分类地位。方法:随机挑选30株本室保存的分离自中国东海的细菌,利用抗大肠杆菌模型进行活性筛选。对活性菌株进行形态特征、生化特征、盐需求测试及16S rDNA序列分析,并将所测得的序列在NCBI数据库进行相似性搜索,选取其中具有代表性的序列以邻接法构建系统进化树,将菌株鉴定到属。结果:筛选出活性菌株F81612和F201721,它们的最适生长盐浓度分别为10%和7.5%,菌株形态特征、生化性质与Bacillus sp.相符,菌株F81612和F201721的16S rDNA序列分别与Bacillus subtilis和Bacillus amyloliquefaciens相似程度最大。结论:利用抗大肠杆菌模型筛选到两株产大环内酯Macrolactin S的细菌,均为中等嗜盐的海洋Bacillus sp.。     

海洋源微生物的分离、鉴定及抑菌与抗肿瘤活性评价

目的从厦门近海采集的海绵样品中分离、纯化及鉴定共附生微生物,并对其进行抑菌与抗肿瘤活性评价。方法根据典型菌落形态特征,共分离纯化得到8株细菌;经16S rDNA鉴定,6株为芽孢杆菌属,1株为变胞杆菌,1株为未分类细菌。结果经纸片法筛查,两株细菌具有较好的应用潜力,其中一株对荧光假单孢菌和白假丝酵母拮抗较强,也具有较好的抗肿瘤活性;另一株则对所测三株指示菌的抑制和抗肿瘤活性都较强。结论筛选出两株兼具抑菌与抗肿瘤活性的海洋源细菌,具有进一步研究应用的价值和潜力。     

中国东海亚硫酸杆菌属微生物的鉴定及生物学活性分析

目的:对44株中国东海来源的亚硫酸杆菌进行16S rDNA的序列测定和生物学活性的初步分析.方法:用PCR方法对细菌的16S rDNA序列进行扩增,并在GenBank上进行相似性搜索.将全部亚硫酸杆菌的16S rDNA序列运用Clustal X1.8软件进行多序列比对并用MEGA4.1绘制进化树.利用稻瘟霉菌(Pyricularia oryzae) P-2b、枯草芽孢杆菌 (Bacillus subtilis) 和大肠埃希菌 (Escherichia coli) 作为指示菌检测菌株发酵液的抗菌活性.以HL-60、HepG2细胞作为细胞毒活性指示细胞株检测菌株发酵液的细胞毒活性.采用ABTS和DPPH抗氧化模型检测菌株发酵液的自由基清除能力.结果:经与GenBank数据库比对,44株菌均与亚硫酸杆菌属有很高的相似性(97%～100%).所有菌株发酵液对指示菌未显示出抗菌活性.菌株发酵液有不同程度的细胞毒活性和清除自由基的能力.19株菌的发酵液对HepG2细胞有抑制作用,抑制率为6.8%～42.8%,18株菌的发酵液对HL-60细胞有抑制活性,抑制率为6.9%～43.6%.12株菌的发酵液对ABTS自由基有清除能力,清除率为4.49%～23.08%,8株菌发酵液对DPPH自由基有清除能力,清除率为1.23%～30.76%.结论:44株中国东海来源的亚硫酸杆菌表现出显著的细胞毒活性及抗氧化活性,具有潜在的药用价值.     

一株高产淀粉酶枯草芽孢杆菌的筛选、鉴定及产酶条件的优化

目的：筛选出α-淀粉酶高产菌株,对其发酵特性及分类鉴定进行研究,为丰富产淀粉酶菌种资源和进一步挖掘该菌株的应用价值奠定基础。方法：通过初筛、复筛获得高酶活菌株XW86,利用传统的形态观察、系统生理生化反应并结合16S rDNA的序列分析,经Blast序列比对构建其系统进化树,确定其分类定位。通过单因素选择实验以及进一步的正交实验和多因素交互实验,进行产酶条件优化。结果：XW86与已报道的枯草芽孢杆菌（Bacillus subtilis strain QD517）亲缘关系最近,二者的16S rDNA序列相似性为99%,生理生化反应谱与16S rDNA序列分析一致鉴定为枯草芽孢杆菌。XW86最适产酶条件为：在LB基础培养基中添加0.75%吐温、0.5%淀粉,初始pH 6.5,35℃培养120 h。液体发酵得到的粗酶液酶活可高达2 200 U/dL,比优化前提高2.8倍。结论：初步鉴定XW86为枯草芽孢杆菌,产酶因素的最佳组合对其淀粉酶活性有很大的优化空间。该野生型菌株可为淀粉酶发酵工业新备选菌株的开发应用提供资源,也为进一步克隆获得产淀粉酶基因的研究奠定基础。     

苯丙氨酸脱氨酶产生菌的筛选、鉴定及酶学性质研究

目的 筛选一株苯丙氨酸脱氨酶高产菌株.方法 从肥沃菜园土样中分离得一株苯丙氨酸脱氨酶产生菌,通过形态、生理生化特性和16S rDNA序列分析,对菌株进行鉴定,并对其酶学性质初步研究.结果 初步鉴定该菌为蜡样芽孢杆菌(Bacillus Cereus),所产苯丙氨酸脱氨酶的最适反应pH5.8,最适反应温度40℃,60℃以下热稳定性较好,此条件下酶活达1 218 U/mg细胞.结论 所获得的蜡样芽孢杆菌NJU110具有一定的苯丙氨酸脱氨酶生产能力,并为该菌株的菌种改良及高效基因工程菌的构建奠定了基础.     

市售川产水豆豉分离株中高产蛋白酶与β-葡萄糖苷酶菌株的菌属鉴定与产酶能力评价

目的 筛选并鉴定72株川产水豆豉分离株中高产蛋白酶及β-葡萄糖苷酶的菌株,为深入研究水豆豉的营养价值及开发其发酵菌株提供实验依据。 方法 采用酪蛋白法和对硝基苯-β-D半乳糖吡喃糖苷(pNPG)法分别测定菌株产蛋白酶和β-葡萄糖苷酶的能力,选出优势菌株,通过形态学、生化特征、16S rRNA和基质辅助激光解析-电离飞行时间质谱(MALDI-TOF-MS)对其进行鉴定。绘制菌株生长曲线和产酶曲线并测得其遗传稳定性。 结果 选择产蛋白酶能力前10名菌株中β-葡萄糖苷酶活力最高者(0.084 U/L)作为优势菌株,表型及分子生物学法鉴定其为枯草芽孢杆菌(Bacillus subtilis)。菌株生长曲线、产酶曲线符合枯草芽孢杆菌特性,遗传稳定性结果显示其产酶能力能稳定遗传至第9代并于第10代开始下降。 结论 从72株川产水豆豉分离株中筛选出的高产蛋白酶和β-葡萄糖苷酶菌株经鉴定为枯草芽孢杆菌。     

16S rDNA序列分析在临床不常见细菌鉴定中的初步应用

目的建立细菌16S rDNA序列分析技术并探讨其在临床不常见细菌鉴定中的应用价值。方法收集经全自动微生物分析系统鉴定其可靠率在99%及以上的常见细菌10株和常规方法难以鉴定或少见菌17株,通过PCR扩增16S rDNA v3-v4区基因片段并进行序列测定,序列与16SpathDB 2.0数据库上已知细菌的16S rDNA序列进行比对分析确定细菌种属。当v3-v4序列片段不能明确鉴定细菌时,结合16S rDNA长片段测序及管家基因dnaJ和rpoB的序列做进一步分析。结果经16S rDNA v3-v4区序列分析,10株(100%)临床常见菌都能鉴定到种水平,其可靠性大于98.0%。17株不常见菌株中,10株(58.8%)细菌只与1种菌相似性大于98.0%,成功鉴定到单个种水平;6株(35.3%)细菌与不止1种菌的相似比大于98.0%;1株菌与已知菌相似性在96.0%~98.0%之间只能鉴定到属(棒状杆菌)。进一步结合16S rDNA长片段及管家基因dnaJ和rpoB的序列分析,所有菌株均可鉴定到单个种水平。结论结合16S rDNA和管家基因序列分析,可用于临床常见及不常见细菌的分子鉴定。     

实验棉铃虫携带微生物的检测

目的 通过对本所长期室内人工饲育的不同季节不同龄期的棉铃虫(Heliothis armigera)进行体内外的微生物分离培养和鉴定,掌握棉铃虫体内外微生物携带情况.方法 利用普通培养基和真菌培养基分别对棉铃虫体内外进行微生物分离培养、纯化,通过菌落形态观察、革兰氏染色和显微镜观察进行初步鉴定,然后应用16SrRNA和18SrDNA分析技术对分离的细菌和真菌进行进一步的鉴定.同时,利用PCR技术对棉铃虫的高度致病病毒-核型多角体病毒(HaNPV)进行扩增检测.结果 分离培养得到4株芽孢杆菌(Bacillus):分别为枯草芽孢杆菌(Bacillus subtilis Cohn),地衣芽孢杆菌(Bacillus licheniformis),蜡状芽孢杆菌(Bacillus cereus)和芽孢杆菌(Bacillus).4株真菌(fungi):其中2株为酵母菌(Yeasts),1株为青霉菌(Aenicil lium Link),1株为曲霉菌(Aspergillus Micheli).检测核型多角体病毒(HaNPV)为阴性.结论 室内人工饲养的棉铃虫生长状态良好,虫体健康,未携带致病微生物,为科研提供的实验昆虫质量稳定可靠.     

两株耐酸耐胆盐人肠源菌株的鉴定及系统发育分析

目的对两株来源于健康人肠道的耐酸耐胆盐菌株R92-2和R111进行鉴定和系统发育分析，以筛选优良的宿主菌株为最终构建能高效表达和分泌β-半乳糖苷酶的工程菌奠定基础。方法表型特征分析后，利用16S rDNA两端的保守序列作为PCR引物，扩增两株菌的16S rDNA序列并测序，测序结果于GenBank中Blastn并构建系统发育树。结果表型特征分析初步鉴定两株菌为乳酸菌。R92-2的16S rDNA序列与Weissella cibaria的16S rDNA序列同源性为100％；R111的16S rDNA序列与Enterococcus faecium的16S rDNA序列同源性为100％。结论R92-2为Weissella cibaria；R111为Enterococcus faeciura。两株菌有望用于构建能高效表达和分泌β-半乳糖苷酶的工程菌。     

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.     

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…     

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.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

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.