RP-HPLC法测定盾叶薯蓣细胞中薯蓣皂甙元的含量

本文采用KromasilC - 18柱 (2 5 0× 4 6mm ,5 μm ) ,甲醇∶水 =95∶5流动相 ,流速 1 0mL/min ,UV检测器 ,检测波长 2 0 8nm ,对培养的盾叶薯蓣细胞中的薯蓣皂甙元含量进行测定 ,能将薯蓣皂甙元与杂质分离开 ,薯蓣皂甙元在 3 876～ 19 38μg范围内峰面积与其浓度线性关系良好 (R =0 9993) ,样品平均回收率为97 83% ,精密度RSD为 0 2 1% (n =4 )。该法测定样品快速准确 ,重复性好     

胃内酸化消除胃内细菌过生长造成的14C-尿素呼气试验假阳性

目的 :观察胃内细菌过生长是否可以引起 1 4C-尿素呼气试验假阳性 ,同时胃内酸化是否能即刻消除这种影响。方法 :5 8名胃黏膜组织学 Hp阴性、常规 1 4C- UBT( U BT- 1)也阴性的溃疡样型功能性消化不良的病人口服雷贝拉唑 2 0 mg/ d、共 7d。于第 6天常规 1 4C- UBT( U BT- 2 )。U BT- 2阳性者于第 7天再进行一次改良的 1 4C- U BT( U BT- 3) ,方法是在服用 1 4C-尿素胶囊前 2 0 m in和服用胶囊时各加饮 15 0 m l0 .1mol/ L的柠檬酸水。比较三次 U BT的 1 4CO2 呼出率。结果 :雷贝拉唑明显增加 Hp阴性病人 1 4CO2 呼出率 ,U BT- 1为 ( 2 4 .0± 10 .8) dpm / mm ol CO2 ,U BT- 2为 ( 6 3.6± 16 .8) dpm / m mol CO2 ( ( t=2 .310 ,P<0 .0 5 )。U BT- 2有 9例假阳性 ( 9/ 5 8,16 % )。但胃内酸化后的 UBT- 3的 1 4CO2 呼出率又落至 ( 2 1.0± 7.8) dpm/mm ol CO2 ( ( t=2 .0 6 8,P<0 .0 5 ) ,9例假阳性者又恢复真阴性。结论 :胃内细菌过生长可以引起 1 4C-尿素呼气试验假阳性 ,同时胃内酸化能即刻消除这种影响     

水氮添加对内蒙古温带典型草原植被氮含量季节和年际动态的影响

作为对全球变化响应最敏感的生态系统类型之一,草原生态系统植被氮含量的季节、年际变化及其对气候变化(氮沉降、降水格局改变)的响应研究相对匮乏。基于内蒙古温带典型草原5年的氮添加(10 g N m~(-2) a~(-1))和水添加(添加量80 mm,分2 mm×40次、5 mm×16次、10 mm×8次、20 mm×4次、40 mm×2次5种处理)控制试验分析了水氮添加后植被氮含量在生态系统和物种两个水平的季节和年际变化。结果表明,高强度水添加处理有降低(10 mm/次和40 mm/次)生态系统氮含量的趋势,但不显著,小强度水添加处理(2 mm/次、5 mm/次)在不同年份之间无一致的升高或降低趋势,但所有水添加处理有降低两种优势物种整个生长季氮含量的趋势。氮添加促进生态系统和两种优势物种整个生长季的氮含量,但该促进作用可被水添加抵消,且这种抵消作用随水氮添加年限的延长而加剧。水氮添加均增加了生态系统氮含量的年际变异,但对特定物种季节内变异的影响在湿润和干旱年份存在一定差异。本研究将为预测草原生态系统对未来氮沉降增加和降水格局改变的响应及模型改进提供科学依据。     

HPLC法测定麻仁润肠丸大黄素、大黄酚的含量

采用KromacilC18(4 .6mm× 2 5 0mm ,5 μm)色谱柱 ,以甲醇 - 0 .1%磷酸溶液 (85∶15 )为流动相 ,流速为 1mL·min ,柱温为 30℃ ,检测波长为 2 5 4nm ,以外标法测定了麻仁润肠丸中大黄素、大黄酚的含量。大黄素在 8.992× 10 -3 ～ 116 .896× 10 -3 ,大黄酚在 2 1.376× 10 -3 ～ 2 77.85 8× 10 -3 范围内呈线性关系 ,其在制剂中的平均回收率 (n =6 )分别为 10 1.5 6 % (RSD =1.3% )、96 .78% (RSD =1.3% )。     

红点锦蛇

红点锦蛇 Elaphe rufodorsata ( Cantor)为无毒蛇。体背面淡红褐色 ,背中央一条 2行鳞片宽的橙黄色的纵线。体侧各有暗黑褐色斑纹连成的纵线二条。头背有“∧”形黑斑。腹面密集不规则的褐黑色小方斑。有些色变异个体其黑色部分转变为褐黄色。上唇鳞 7( 8) ,2 ( 3) - 2 - 3式 ,眼前鳞 1 ( 2 ) ,眼后鳞 2 ,颞鳞 2 ( 3,1 ) 3( 2 ,4)。背鳞平滑 ,2 1 ( 2 3) - 1 9( 2 1 ) - 1 7( 1 5)行。腹鳞雄性 1 63～ 1 86,雌性 1 68～ 1 88;尾下鳞雄性 51～ 64,雌性 47～ 56。体长雄性可达 843 1 44mm,雌性可达 71 3 1 0 8mm。该蛇为半水生性 ,常在田…     

半制备高效液相色谱制备5种苯乙醇苷类单体

利用MCI柱层析和半制备型反相高效液相色谱法从牛耳朵中分离制备了5种苯乙醇苷类单体。在Eclipse XDB-C18(9.4mm×250mm,5μm)半制备柱上,以甲醇-水为流动相,流速为4mL/min,采用梯度洗脱方式,制备得到5个组分。经波谱(EI-MS、1H-NMR、13C-NMR)分析,分别鉴定为plantainoside(1)、chirito-sideC(2)、plantaninosideB(3)、plantamajoside(4)和desrhamnosylverbascoside(5)。高效液相色谱分析表明所制备5种化合物纯度均高于98%。该方法分离条件简单,高效,一次可得到5个单体,目标产物纯度较高。     

西红花的离体成花

1植物名称西红花(Crocus sativus). 2材料类别球茎的幼嫩顶芽,长度在20～30 mm. 3培养条件诱导培养基分别为:(1)MS NAA 5mg·L-1(单位下同) 6-BA 5;(2)MS NAA 5 6-BA7;(3)MS KT 5 NAA 4;(4)MS 2,4-D 2 KT0.5;(5)MS 6-BA 2 2,4-D 0.1;(6)MS 6-BA2 IBA 1;(7)B5 KT 5 IAA 4 2,4-D 1 Pro 0.5;(8)B5 KT 5 IAA 4 2,4-D 1 Pro 0.5.以上培养基均添加琼脂0.8%;(1)～(5)中加蔗糖4%,(6)中3%,(7)中6%,(8)中4%;pH 5.8.培养温度(20±2)℃,黑暗培养.     

云南姜花属─新种

Hedychium menglianense Y. Y. Qian sp. nov. (Subgen. Euosmianthus)Species affinis H. sino-aureo Stapf, sed plantis ad arbores epiphyticis, laminis (2--)10--27 cm longis, (0.5 -) 1 .7-4 cm latis, petions 2-8 mm longis, ligulis papyraceis,l.5 -2 mm longis, spicis 10- 17-floribus, 5 -- 8 cm longis, bracteis calycis tubos noncircumnexis, apice emarginatis, calycis tubis bractea parum brevioribus, corollae lobislinearibus 11-13 mm longis, labellis ovatis, 8-9 mm longis, basi sessilibus, filamentis…     

四爪陆龟血细胞形态学参数的初步研究

对四爪陆龟的血细胞进行了显微观察 ,并对红细胞和白细胞及其细胞核的大小和面积、血红蛋白含量以及红细胞渗透脆性等参数进行测量。结果表明 :( 1 )四爪陆龟红细胞含量平均值为 62 ( 5 2～ 83 )万个 mm3 血液 ;( 2 )红细胞的平均长度为 ( 1 6 70± 1 2 5 ) μm ,范围 1 5 60～ 1 7 5 2 μm ;( 3 )红细胞宽度为 ( 9 5 3± 0 92 ) μm ,范围 7 2 0～ 1 0 8μm ;( 4 )红细胞核长度为 ( 5 5 9± 0 5 9) μm ,范围 4 80～ 6 48μm ;( 5 )红细胞核宽度为 ( 3 65± 0 5 6) μm ,范围 2 64～ 4 5 6μm ;( 6)血红蛋白含量为 ( 7 3 0± 0 66)g 1 0 0ml血液 ( 6 5 0～8 5 0 ) ;( 7)红细胞渗透脆性 ,开始溶血浓度为 0 2 4%NaCl溶液 ,完全溶血浓度为 0 1 6%NaCl溶液 ;( 8)红细胞平均大小 (长径×短径 )为 ( 1 6 70× 9 5 3 ) μm2 ;( 9)白细胞平均值 0 5 6( 0 42～ 0 70 )万 mm3 ,其中嗜中性粒细胞大小为 ( 1 2 0 8± 1 86) μm ,嗜酸性粒细胞大小为 ( 9 2 5± 2 3 ) μm ,嗜碱性粒细胞大小为 ( 8 75±0 2 3 ) μm ,单核细胞大小为 ( 1 1 40± 1 0 6) μm ,淋巴细胞大小为 ( 7 5 0± 2 1 ) μm ,凝血细胞大小为 ( 4 2 0±1 40 ) μm。     

自然光照下普陀水仙的组织培养

1　植物名称　水仙 (Narcissustazettavar.chi nensis)品种普陀水仙。2　材料类别　带鳞茎盘的双鳞片 (twinscale)。3　培养条件　以MS为基本培养基。 ( 1 )诱导小鳞茎培养基 :MS + 6 BA 5mg·L- 1 (单位下同 ) +NAA 0 .2 5～ 0 .5 ;( 2 )生根培养基 :1 /2MS(大量元素减半 ) +NAA 0 .0 5。培养基均加 0 .8%琼脂 (a gar) ,( 1 )加0 .5 %活性炭 (activatedcharcoal)和 4%蔗糖 (sucrose) ,( 2 )加 3%蔗糖 ,pH 5 .6～ 5 .8。培养温度 ( 2 5± 2 )℃ ,置于培养室自然光线下 ,不用灯光照明。4　生长与分化情况4.1　无菌材料的获得　 5、6月…     

血清CEA、NSE、CYFRA21-1 联合支气管活检在周围型肺癌诊断鉴别中的应用

目的:探讨血清肿瘤标志物癌胚抗原(CEA)、神经元特异性烯醇化酶(NSE)、细胞角蛋白19片段(CYFRA21-1)联合经纤维支气管镜肺活检(TBLB)在周围型肺癌诊断鉴别中的应用价值。方法:选择2012年9月到2014年9月在我院临床确诊的402例周围型肺癌患者。检测所有患者的血清CEA、NSE和CYFRA21-1的浓度及分析TBLB检测结果,分析不同病理类型和不同病灶直径大小时各项检测指标及联合检测的阳性检出率。结果:所有肺癌患者中,血清CEA、NSE、CYFRA21-1和TBLB的阳性检出率分别为51.74%,35.07%,41.79%和60.70%。四项指标联合检测的阳性检出率为89.05%,明显分别高于四项指标的阳性检出率(均P0.05)。腺癌、鳞癌、小细胞肺癌及其他类型肺癌组四项联合检测的阳性检出率明显高于四项单独检测(均P0.05)。病灶直径为2 cm、2-6 cm和6 cm时,四项联合检测的阳性检出率明显高于四项单独检测(均P0.05)。结论:血清CEA、NSE、CYFRA21-1联合TBLB检测周围型肺癌较单项检测,阳性检出率高,值得在临床上推广应用。     

Specific and sensitive detection of Ralstonia solanacearum in soil with quantitative, real-time PCR assays

Aims:  The aim of this study was to develop a sensitive and an effective method suitable for large-scale detection and quantification of Ralstonia solanacearum in soil.
Methods and Results:  Based on the specific sequence of R. solanacearum strain G1000, the primer pair R.sol1-R.sol2 and the TaqMan probe Rs-pro were designed, and specific and sensitive PCR detection methods were successfully established. The detection limit was 100 fg μl⁻¹ DNA in conventional PCR and 1·2 fg μl⁻¹ in real-time PCR. By combining real-time PCR with the modified protocols to extract DNA from soil, it was possible to achieve real-time detection of R. solanacearum in soil, and the degree of sensitivity was 100 fg μl⁻¹. To detect inhibition in soil samples, an exogenous internal positive control (IPC) was included preventing false negative results, and IPC was successfully amplified from all samples tested. The methodology developed was used to detect the presence of R. solanacearum in tobacco fields in China.
Conclusions:  The real-time PCR combined with the protocol to extract DNA from soil led to the development of a specific, sensitive and rapid detection method for R. solanacearum in soil.
Significance and Impact of the Study:  The real-time PCR improves the detection sensitivity and specificity and provides an important tool for routine detection of R. solanacearum in soil samples and for epidemiological and ecological studies.     

The development of rapid real-time PCR detection system for Vibrio parahaemolyticus in raw oyster

Aims:  To develop a new rapid real-time polymerase chain reaction (PCR) based detection system for Vibrio parahaemolyticus ( V. parahaemolyticus ) applicable to raw oyster samples.
Methods and Results:  V. parahaemolyticus cells were artificially inoculated to oysters. Samples were homogenized in 100 ml of sterile saline water and serially diluted to 1·5 CFU ml⁻¹ level. One millilitre of diluents was centrifuged and the pellet was resuspended with 100  μ l of de-ionized water. DNA was extracted by boiling for 20 min, and 0·5  μ l was used as a template for PCR reaction. Real-time PCR was performed with TMC-1000 system (1  μ l PCR system). The detection system was found to achieve detection limit of 1·5 CFU g⁻¹ for V. parahaemolyticus . Furthermore, the specificities of these assay systems were confirmed with more than 20 bacterial strains, including various Vibrio species.
Conclusions:  Rapid and sensitive food-borne pathogen detection techniques for V. parahaemolyticus is important to the food industry and consumers. The direct detection of V. parahaemolyticus from food is possible with micro real-time PCR system.
Significance and Impact of the Study:  This study shows that oyster samples can be tested for V. parahaemolyticus with a rapid, specific and simple procedure.     

Assessment of the stability of human viruses and coliphage in groundwater by PCR and infectivity methods

Aim: To investigate the potential health hazard from infectious viruses where coliphages, or viruses by polymerase chain reaction (PCR), have been detected in groundwater. Two aspects were investigated: the relationship between infectivity and detection by PCR and the stability of coliphage compared to human viruses. Methods and Results: Virus decay (1 year) and detection (2 years) studies were undertaken on groundwater at 12°C. The order of virus stability from most to least stable in groundwater, based on first‐order inactivation, was: coliphage ΦX174 (0·5 d^?1) > adenovirus 2 > coliphage PRD1 > poliovirus 3 > coxsackie virus B1 (0·13 d^?1). The order for PCR results was: norovirus genotype II > adenovirus > norovirus genotype I > enterovirus. Conclusions: Enterovirus and adenovirus detection by PCR and the duration of infectivity in groundwater followed similar trends over the time period studied. Adenovirus might be a better method for assessing groundwater contamination than using enterovirus; norovirus detection would provide information on a significant human health hazard. Bacteriophage is a good alternative indicator. Significance and Impact of the Study: PCR is a useful tool for identifying the health hazard from faecal contamination in groundwater where conditions are conducive to the survival of viruses and their nucleic acid.     

血清MCP-1、RANTES及CysC联合检测对急性冠状动脉综合征的诊断价值

目的:探讨联合检测血清单核细胞趋化蛋白-1(MCP-1)、活化T细胞趋化因子(RANTES)及半胱氨酸蛋白酶抑制剂C(Cys C)对急性冠状动脉综合征(ACS)患者的早期诊断和预后评估价值。方法:选择200例于2012年10月到2013年10月在本院就诊的冠心病患者200例,其中ACS患者120例(为ACS组),另稳定型心绞痛(SAP)患者80例(SAP组),另选择同期40例健康人为对照组。115例患者有冠状动脉粥样硬化,其中40例钙化斑块组、42例纤维斑块组及33例软斑块组,进行CT检查;用双抗体夹心酶联免疫(ELISA)法检测患者血清MCP-1、RANTES、Cys C及高敏C反应蛋白(Hs-CRP)水平,并用logistic回归方程评价联合检测MCP-1、RANTES及Cys C预测ACS的效果。结果:与SAP组、对照组相比,ACS组患者MCP-1、RANTES、Cys-C、Hs-CRP、LDL-C及空腹血糖(FBG)浓度均明显升高(P0.05);HDL-C浓度明显降低(P0.05);与纤维斑块组、钙化斑块组相比,软斑块组MCP-1、RANTES及Cys-C浓度均明显升高(P0.05);与钙化斑块组相比,纤维斑块组MCP-1、RANTES及Cys-C浓度均明显升高(P0.05);联合检测血清MCP-1、RANTES、Cys-C预测ACS患者,阳性准确率为89.2%,阴性准确率为92.5%,综合准确率为90.8%,明显高于单纯Hs-CRP的检测方法(P0.05)。结论:血清MCP-1、RANTES及Cys C联合检测对急性冠状动脉综合征的诊断效果优于单纯检测Hs-CRP。该联合检测方法具有一定的临床应用价值。     

新疆维吾尔自治区20年间胃癌的临床特点分析

目的:分析新疆地区近20年来胃癌流行病学特征,探讨其变化规律及发展趋势。方法:回顾性分析和比较1991年、2001年、2011年经新疆维吾自治区人民医院胃镜及病理学诊断确诊为胃癌的病例的一般资料、病理学及内镜下特点,包括性别、年龄、病理类型、发病部位。结果:1991年组:胃癌检出率为2.48%,中位年龄为54岁,男女之比为3.3:1.0,发病部位以胃窦部癌为主,占39.1%;2001年组:检出率为2.39%,中位年龄为61岁,男女之比为3.0:1.0,发病部位以胃体部癌为主,占42.1%;2011年组:检出率为1.48%,中位年龄为63岁,男女之比为3.9:1.0,发病部位以贲门胃底部为主,占34.8%。三组病理学类型均以腺癌为主,检出率有逐年升高趋势,但差异无统计学意义(P0.05)。结论:(1)近20年来胃癌发病部位有上移现象,且胃癌检发病率有下降趋势;(2)男性胃癌患者发病率明显高于女性,且近20年来胃癌患者男女比例无明显改变;(3)近20年来胃癌发病中位年龄逐渐增高,且随着年龄的增长发病率逐渐升高,以中老年发病率最高;(4)癌患者病理类型仍以腺癌为主,且近20年来腺癌所占比例无明显变化。     

AFP蛋白芯片技术的检测流程优化

目的:通过优化现有的蛋白芯片检测过程,在保证检测准确性的同时缩短甲胎蛋白(Alpha Fetal protein,AFP)的检测时间,提高检测效率,为原发性肝癌的筛查提供经济、便捷、省时、有效的检测方法。方法:本研究在传统蛋白芯片检测流程(1-1.5小时)的基础上,通过优化检测流程将检测时间缩短至18分钟,并且通过和传统方法进行比较,评价该优化方法的检测效能。结果:与传统蛋白芯片检测方法相比,本优化方法的检测时间缩短至18分钟。重复检测同一样本,传统方法 AFP水平为16.50±1.172ng/m L,优化方法 AFP水平为18.33±1.029 ng/m L,结果无明显统计学差异(P=0.251)。结论:本研究成功地优化了AFP的蛋白芯片检测流程,在缩短检测时间的同时,保证了检测的准确率,是一种经济省时易操作的AFP检测方法。     

Trials of direct detection and identification of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG 1 and AG 2 subgroups using specifically primed PCR analysis

Specifically primed polymerase chain reaction (PCR) analysis was used for direct detection and identification of Rhizoctonia solani isolates belonging to AG 1 subgroups (IA, IB, and IC) and AG 2 subgroups (2-1 and 2-2). A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract PCR templates. PCR-specific primer sets for each group were designed from sequences in the regions of the 28S ribosomal DNA of this fungus. The results of specifically primed PCR analysis showed that AG 1-IA, AG 1-IB, AG 1-IC, AG 2-1, and AG 2-2 primers sets contributed detection from the same AG isolates and could escape detection from different AG isolates at a high level of frequency. In this experiment, we suggested that our synthesized primer sets might provide a method for the direct detection and identification of AGs of R. solani. Received: June 28, 2001 / Accepted: November 14, 2001     

糖化血红蛋白在糖尿病中的应用价值分析

目的:研究糖化血红蛋白的检测在糖尿病患者中的诊断价值及对代谢指标的影响。方法:对126例临床确诊为糖尿痛患者进行糖化血红蛋白及空腹血糖检测,并且将糖化血红蛋白分为A组(HbA1c≤10%)和B组(HbA1c>10%),比较两组TG,TC,LDL.HD等指标。结果:FPG在B组患者的水平明显高于A组患者的水平,差异有显著的统计学意义(P<0.01),而2hPG,TC,LDL B组患者的水平明显高于A组患者的水平,差异有明显的统计学意义(P<0.05)。结论:检测糖化血红蛋白在糖尿病患者具有重要的临床价值。     

The diagnostic value of combined detection of genetic markers and serum protein markers on breast cancer

Objective: The research is to explore the diagnostic value of several detection methods including separated and combined detection of the related genes and related proteins of breast cancer and combined detection of all genetic markers and serum protein markers on breast cancer. Method: The mRNA level expression of the related genes of breast cancer was detected by FQ-PCR technique and the ratio of BRCA-1, Myc, C-erbB2 and β2 micro-globulin was used to express levels of BRCA-1, Myc and C-erbB2; the related proteins of breast cancer were detected through ELISA. Then the research data was analyzed by SPSS19.0 software with t-test as comparison method, and ROC curve was used to calculate the sensitivity, specificity and accuracy of the diagnostic models. Result: No difference can be found among the six indexes in the control group and benign breast tumor group while compared with the benign breast tumor group and the control group, the breast cancer group was significantly different from them; combined detection of genes and that of proteins were both superior to their separated detection; all-marker combined detection was superior to separated detection, which is consistent with combined detection of genes and proteins. Conclusion: More detection indexes will not necessarily outcome better detection effect. Hence, appropriate detection indexes and number are needed to achieve better diagnosis effect. In order to conduct more specific method, more test samples are needed for further researches.