RP-HPLC法测定丹参药材中丹参素及原儿茶醛含量

本文建立了用于测定丹参药材中丹参及原儿茶醛的反相高效液相色谱(RP-HPLC)分析法。方法采用以甲醇和0.25％醋酸溶液分别为流动相A和B的梯度洗脱技术,YWGG_(18)H_(37)色谱柱,UV280nm检测波长以及内标化合物对羟基苯甲酸,丹参药材先以水为溶剂的索氏提取法提取,再将提取液制成供试液。试验结果示分离完全的色谱峰,良好的线性相关(γ>0.99)和满意的回收率(丹参素为104.04±1.7％,原儿茶醛为104.6±2.9％)。按本法测得不同产地丹参药材中的含量之间有明显的差异,而饮片中的含量则均在不同程度上低于原药中的含量,提示饮片切制过程中丹参素及原儿茶醛有不同程度的损失。

高效液相色谱法测定香丹注射液中丹参素和原儿茶醛的含量

目的 建立反相高效液相色谱法测定香丹注射液中丹参素和原儿茶醛的含量。方法 以对羟基苯甲酸为内标 ,色谱柱 :ALL TECH ODS柱 (2 5 0× 4.6mm,5μm) ;流动相 :水 -乙腈 -冰醋酸 ;流速 :1.2 ml· min- 1 ;检测波长 :2 81nm。 结果 丹参素加样回收率为 97.65 %～ 10 0 .10 % ,RSD为 0 .7%～ 1.2 % ;原儿茶醛加样回收率为 97.5 6%～ 10 0 .3 0 % ,RSD为 1.0 %～ 1.6% ,(n=5 )。结论 实验结果表明 ,该方法操作简便 ,重现性好 ,可作为样品的检测方法。

RP-HPLC法测定资木瓜中原儿茶酸的含量

目的:建立以RP-HPLC法测定资木瓜中原儿茶酸含量的方法。方法:色谱柱:Agilent Eclipse XDB-C_(18)(250 mm×4.6mm,5μm),流动相:乙腈-0.1%磷酸水溶液(5:95),流速1 ml·min^(-1),检测波长260 nm。结果:原儿茶酸进样量在0.02～0.20μg范围内具有良好的线性关系(r=0.9999),平均加样回收率为101.23%,RSD为1.83%(n=5)。结论:本方法简便、准确、重复性好,可用于控制资木瓜的质量。

HPLC法测定血尿胶囊中原儿茶酸的含量

目的建立了高效液相色谱法测定中药制剂血尿胶囊中原儿茶酸含量的方法。方法样品用乙醇-醋酸(10:1)超声进取,甲醇溶解,以0.05%磷酸-甲醇(80:20)为流动相,275nm下检测。结果平均回收率98.34%。结论本方法灵敏、准确、专属性好,可作为该制剂的含量测定及质量控制。

HPLC法测定心舒宝胶囊中原儿茶醛的含量

目的:测定心舒宝胶囊中原儿茶醛的含量。方法:采用十八烷基硅烷键合硅胶柱;流动相为甲醇-0.5%冰醋酸溶液(20:80)为流动相,检测波长为280nm;速为1.0ml/min;结果:原儿茶醛线性范围5.05～50.5μg,平均回收率为98.8%,RSD=0.6%(n=6);结论:该方法不仅操作简单,准确,且重现性好,可用于该制剂的质量控制。

川芎嗪-原儿茶酸络合物的合成和其抗缺血性脑损伤作用初步研究

目的:以川芎嗪、原儿茶酸为前体原料,运用药物拼合理论,合成川芎嗪-原儿茶酸络合物,并对其进行初步体内外缺血性脑损伤活性研究。方法:以川芎嗪(Tetramethylpyrazine,TMP)和原儿茶酸(Protocatechuic acid,PCA)为原料合成川芎嗪-原儿茶酸络合物(Tetramethylpyrazine-Protocatechuic acid,TP),运用UV、1H-NMR、13C-NMR、MS、X-Ray确定该化合物的结构。通过①鸡胚绒毛尿囊膜血管生成(Chicken chorioallantoic membrane,CAM)模型观察TP对新生血管生长的影响;②MTT法测定TP对原代培养的正常胎鼠皮层神经细胞、谷氨酸(Glutamate,Glu)和NO损伤神经细胞模型的影响;③TP对小鼠断头张口呼吸时间的影响等实验初步观察其对缺血性脑损伤的拮抗作用。结果:确定TMP和PCA两个分子片断之间借助一个O-H…O氢键和一个O-H…N氢键连接;TP对新生血管的生长具有显著的促进作用,改善Glu、NO对神经细胞的损伤,并能显著延长小鼠断头后张口呼吸时间。结论:由TMP和PCA拼合而成的TP对缺血性脑损伤具有明显改善作用,显示了其良好的新药开发研究前景。

HPLC法同时测定复方儿茶酊中儿茶素与原儿茶素的含量

目的:建立同时测定复方儿茶酊中儿茶素与原儿茶素含量的方法。方法:采用高效液相色谱法。色谱柱为YMC ODS(250mm×4.6mm,5μm),流动相为0.04mol·L-1枸橼酸溶液-N,N-二甲基甲酰胺-四氢呋喃(45∶8∶2,V/V/V),流速为1.0mL·min-1,检测波长为280nm,柱温为35℃。结果:儿茶素与原儿茶素的检测浓度分别在0.0518～0.2590、0.0101～0.2030mg·mL-1(r均为0.9999)范围内与各自峰面积积分值呈良好的线性关系;平均加样回收率分别为101.63%和100.49%,RSD分别为1.27%和1.12%(n均为9)。结论:本方法简便、准确、稳定,能有效控制复方儿茶酊的质量。

原儿茶酸调控PPARγ通路对小鼠胆总管梗阻肠屏障损伤的保护作用

目的:研究原儿茶酸对小鼠胆总管梗阻肠屏障损伤的保护作用及作用机制。方法:构建小鼠胆总管结扎(BDL)术继发小肠屏障损伤模型。将小鼠随机分为假手术组、模型组、模型+原儿茶酸低、中、高剂量(20 mg/kg、40 mg/kg、80 mg/kg)组,各给药组于术前2天给至术后5天腹腔注射相应剂量原儿茶酸。第5天末处死小鼠,检测小肠形态学、小肠PPARγ通路相关氧化应激和细胞凋亡指标及全身炎症指标;利用Western blot检测小肠PPARγ、Occludin和Bcl-2蛋白表达。结果:模型组小鼠较对照组病理损伤评分显著升高,血清TNF-α、IL-6和TB显著升高,小肠组织SOD、GSH、GSH-Px降低,MDA升高;小肠细胞凋亡显著。原儿茶酸腹腔注射原儿茶酸20 mg/kg、40 mg/kg、80 mg/kg组可不同程度改善上述指标变化,并呈剂量依赖趋势。模型组较对照组小肠PPARγ、Occludin、Bcl-2表达降低;原儿茶酸处理组较模型组可显著提升上述蛋白在小肠组织中的表达。结论:原儿茶酸能在小鼠胆总管梗阻肠屏障损伤中发挥保护作用,其机制可能与提高小肠PPARγ表达,减轻小肠组织的氧化应激和细胞凋亡相关。

The Chemical Constituents from the Bulbs of Ornithogalum caudatum

A new natural product (1) together with 26 know compounds were isolated from the Bulbs of Ornithogalum caudatum. Their structures were established on the basis of spectral analyses as n-butyl pyroglutamate (1), nonadecyl alcohol(2), eicosanol(3), behenic acid(4), b-sitosterol(5), stigmasterol(6), glycerol 1-monocerotate(7), pyrocatechol(8), p-ethoxybenzoic acid(9), p-coumarinic acid(10), protocatechuric acid(11), ursolic acid(12), betulinic acid(13), fumaric acid(14), succinic acid(15), uracil(16), xanthine(17), quercetin(18), kaempferol (19), isorham-netin(20), adenosine(21), daucosterol(22), stigmasterol 3-O-b-D-glucopyranoside(23), quercetin 3-O-b-D-glucopyra-noside(24), kaempferol 3-O-b-D-glucopyranoside(25), rutin(26), and kaempferol 3-O-b-rutinoside(27). All of them, except compound 5, were isolated from this plant for the first time.

Effects of Water─soluble Components Isolated from Salviamilltiorrhiza on Oxygen Free Radical Generation and LipidPeroxidation

The effects of water─soluble components isolated from Salvia miltiorrhiza on oxygen free radical prodction and lipid peroxidation were estimated. Five components(10 μmol/L)were shown to inhibit superoxide anion generation by xanthine─xanthine oxidase system. It was also demonstrated that six components( 100 μmol/L) prevented H_2O_2─induced hemolysis and MDA fonnation in mouse erythrocytes.The effects of Sal A, Sal B, Sal C and Ros A were dose dependent. In Langendorff rat heart, pretreatment with Sal A 20μmol/L significantly prevented MDA production induced by 30 min reoxygenation after 45 min anoxia.The results indicated that there were also other potent antioxidant componentts in Salvia milliorrhiza besides Dphl and Pal. The protection of Sal A against myocardial anoxiareoxygenation inury may be mainly ascribed to its oxygen free radical scavenging activity.

基于HPLC法研究广藿香中加入陈皮后水溶性成分的变化规律

采用加热回流法提取广藿香药材的水溶性成分,建立高效液相色谱法(High Performance Liquid Chromatography,HPLC),测定广藿香药材中加入陈皮药材后其水溶性物质(香草酸、原儿茶酸)含量的变化。使用超快速液相色谱法对广藿香化学成分中的水溶性物质的线性关系进行了考察,分别测定了两种提取液中香草酸和原儿茶酸的含量。对广藿香-陈皮混合药材进行提取得到的原儿茶酸的含量比配伍前调高了7倍,香草酸的含量提高了2倍,加入陈皮后广藿香的两种水溶性成分的含量均有所增长。实验的重复性也较好,可对加入陈皮后的广藿香水溶性成分提取效率变化的进一步研究提供参考。

基于HPLC法对加入厚朴后广藿香水溶性物质的含量影响研究

建立高效液相色谱法测后对其广藿香加入厚朴两种水溶性成分香草酸和原儿茶酸的含量影响。采用蒸馏水为溶剂的加热回流法进行提取,广藿香与厚朴配伍后原儿茶酸含量是配伍前的1.6倍,香草酸含量升高明显,配伍后香草酸含量约是配伍前的15.9倍。采用HPLC法对广藿香加入厚朴前后的两种水溶性成分(香草酸,原儿茶素酸)进行测定,该方法科学易行、操作简单、结果准确。

Transformation of hepatitis B serologic markers in babies born to hepatitis B surface antigen positive mothers

AIM:To better understand the clinical significance of hepatitis B seroiogic markers in babies born to hepatitis B surface antigen (HBsAg) positive mothers, the incidence of maternal seroiogic markers of hepatitis B via placenta and its transformation in these babies were investigated. METHODS: Mothers with positive HBsAg were selected in the third trimester of pregnancy. Their babies received immunoprophylaxis with hepatitis B immunoglobulin and hepatitis B vaccine after birth, and were consecutively followed up for hepatitis B seroiogic markers and HBV DNA at birth, mo 1, 4, 7, 12, and 24. RESULTS: Forty-two babies entered the study, including 16 born to hepatitis B e antigen (HBeAg)-positive HBsAg carrier mothers and 26 to HBeAg-negative HBsAg carrier mothers. Apart from four babies born to HBeAg-positive carrier mothers and demonstrated persistent positive HBeAg eventually became HBV carriers, all other babies developed anti-HBs before 12 mo of age. Among the other 12 babies born to HBeAg-positive carrier mothers, HBeAg was detected in 7 at birth, in 4 at mo 1, and in none of them thereafter. No antibody response to the transplacental HBeAg was detected. Among the babies born to HBeAg-negative carrier mothers, anti-HBe was detected 100% at birth and mo 1, in 88.5% at mo 4, in 46.2% at mo 7, in 4.2% at mo 12 and none in mo 24. Among all the immunoprophylaxis-protected babies born to either HBeAg-positive or HBeAg-negative carrier mothers, anti-HBc was detected in 100% at birth, mo 1 and mo 4, in 78.9% at mo 7, in 36.1% at mo 12 and in none at mo 24. CONCLUSION: HBeAg can pass through human placenta from mother to fetus and become undetectable before 4 mo of age, but no antibodies response to the transplacental HBeAg can be detected till mo 24 in the immunoprophylaxis-protected babies. The sole existence of anti-HBe before 1 year of age or anti-HBc before 2 years of age in babies born to HBsAg carrier mothers may simply represent the transplacental maternal antibodies, instead of indicators of HBV infection status.

Low Blood Lead Concentrations and Cognitive Development of Primary School Children from Three Areas in Malaysia

A study on to identify the relationship between blood lead and cognitive development was conducted on primary school children in Malaysia. About 413 children aged 62^/1 - 82^/1 years from urban （236）, rural （80） and industrial areas （97） were studied. Blood lead was analyzed using the Atomic Absorption Spectrophotometer （GFAAS）. Cognitive development was measured by the ＇McCarthy Scales of Children＇s Abilities Test＇ （MSCA）. Significant differences found in the mean cognitive scores between the urban （94.40）, industrial （102.90） and suburban children （101.24） （p 〈 0.001）, with the blood lead between the urban （3.66 p.g/dL）, industrial （3.54 μg/dL） and suburban children （3.04μg/dL） （p = 0.022）. Significant inverse correlations between blood lead and cognitive scores found for all groups （p 〈 0.001）, urban （p 〈 0.001） and suburban children （p 〈 0.001）. Low blood lead significantly influenced the cognitive development for all children after adjusting for confounders （p _〈 0.001）. The urban children＇s cognitive scores were significantly influenced by blood lead levels and household income （p 〈 0.001）. However, for the suburban children, the cognitive score were significantly influenced by the blood lead levels, the mothers＇ education, number of siblings, sequence in the family and the household income （p 〈 0.001 ）. Blood lead below 10μg/dL influenced the cognitive development. Urban children had higher blood lead but suburban children with lower blood lead were also vulnerable to the effect of lead on their cognitive development.

Analysis of Serum Levels of IgA Antibodies to Epstein Barr Virus Capsid Antigens in the Spouses and the Children of Patients with Nasopharyngeal Carcinoma

OBJECTIVE To analyze the serum levels of IgA antibodies to Epstein Barr virus capsid antigens （EBV/IgA/VCA） in patients with nasopharyngeal carcinoma （NPC） and in their spouses and children in order to further evaluate the risk of developing the disease in family members of NPC patients. METHODS Four categories of sera were used to detect EBV/ IgA/VCA using the immunoenzyme method. In our study 317 biopsy-confirmed NPC patients, 317 spouses and 317 children of the NPC patients, as well as 413 healthy subjects as the controls that came from the same area were enrolled. RESULTS The positive rate of EBV/lgA/VCA was 97.2%, 14.2%, 19.9% and 3.1% in the NPC patients, the spouse and child groups, and in the control group, respectively. The positive rate was significantly higher in the NPC group than in the other 3 groups, and it was also significantly higher in the spouse and in the child groups than in the control group （P 〈 0.001）. The results of the relative to identified distribution unit （Ridit） analysis showed that the average Ridit values were 0.860, 0.404, 0.424 and 0.356 respectively in the NPC patients, in the spouse and child groups, and in the control group. The antibody titer of IgA/VCA was significantly higher in the NPC group than in the other 3 groups, and it was also significantly higher in both the spouse and child groups than in the control group （P 〈 0.001）. The OR values of positive EBV/IgA/VCA antibody were 5.09 and 7.63, respectively in the spouse and child groups. No significant differences were found in the positive rates or in the titers of IgA/VCA antibody between the groups of spouses and children （P 〉 0.05）. CONCLUSION Positive detection of EBV/IgA/VCA antibody occurs in familial aggregation, as there is ample opportunity for EBV reactivation in the spouses and in the children of NPC patients. These individuals with a high risk of developing the NPC should be closely followed in order to detect the disease at an early stage.

THE STUDY ON RELATED GENES IN THE NEOPLASTIC TRANSFORMATION OF IMMORTALIZED HUMAN FETAL TRACHEAL FIBROBLAST CELLS INDUCED BY IRRADIATION

In this study, we investigated the genes related to the transformation of immortalized human fetal tracheal fibroblast cell line induced by alpha particles by means of differential display mRNA method. The result revealed that there were 23 DNA fragments that were expressed intensively in alphaSHTF cells (SHTF cells forming clone on agar after irradiated by alpha particles emitted by 238Pu) only and not in SHTF (SV40-immortalized human fetal tracheal fibroblast) cells. Northern dot confirmed two fragments, C17-5, C23-1 which showed intensive mRNA expression in alphaSHTF cells, but not in SHTF cells. The length of the C17-5 fragment was 310bp. Searching in BLAST database revealed that the C17-5 fragment might be an unknown sequence.

The causes of pre-primary school and strategies of removal of pre-primary school

Psychological and physiological characteristics of preschool children determine their lives should be the main game, but because of the content and the way too pre-primary school of teachers and parents give their children pressure of learning early, they often give children physical and cause a series of adverse consequences of psychological, so that children lively nature is destroyed. In the development context of pre-primary schools it will not only hinder enhance children＇ s thinking ability, but also devoid of the child＇ s talent and mettle, for children to learn and live very unfavorable late. Preschool Primary School, out of the office park on the right track, is serious impediment to the normal development of preschool children and grow. This paper describes the dangers of pre-school, pre-primary schools and the causes are analyzed and we proposed the removal of the primary effective strategy, in order to fully mobilize the child＇ s lively and good mobility and lay its future development a solid foundation.

Submicroscopic Plasmodial Infection May Lead to Severe Malaria in Children

Malaria is one of the leading causes of consultation in African pediatric hospitals. In Gabon, malaria transmission is perennial. Plasmodium falciparum, responsible for the most severe form of the disease, represents more than 95% of all species. In P. falciparum infection, the hyperparasitemia accounts among the main criteria of disease severity. However, in many endemic countries, a significant decrease of malaria burden accompanying with the diminution of parasite load in infected individuals has been demonstrated. The objective of the study was to analyze the occurrence of febrile syndrome in childhood and investigate whether the acute febrile illness could be associated with P. falciparum submicroscopic infection. A cross-sectional study was carried out during January to March 2013 in Franceville. A total of 203 acute febrile children were enrolled. A clinical examination and biomedical analysis including parasitological diagnosis by microscope were carried out in all the patients and PCR on microscope negative ones. Of 203 children recruited for febrile syndrome, 111 have been diagnosed positive for P. falciparum infection, 73 （35.9%） by microscope （ME） and 38 （18.71%） by PCR （submicroscopic infection = SM1） with an overall prevalence of 54.68%. Of the 11 1 P. falciparum infected individuals and according to the WHO criteria, 35 （31.53%） children showed a clinical picture of severe malaria against 76 （68.47%） others classified as uncomplicated malaria. The overall prevalence rates were therefore estimated as 17.24 （35/203） for severe cases and 37.43% （76/203） for uncomplicated ones. Clinically, these severe malaria cases （27 ME＋ and 8 PCR＋） were mainly composed of 85.71% of anaemic patients （30/35）, 71.14% of prostrated individuals （25/35） and 57.14% of children with clinical icterus （20/35）. However, only two cases of severe anaemia were observed, the remaining others cases were moderate （10） and mild anaemia （18）. More interestingly, eight submicroscopic infected patients （22.85%） were found with neurological manifestations （prostration） and all were experiencing thrombocytopenia. Lastly, 1 hyperparasitemia, 6 hypoglycemia and 2 respiratory distresses were also observed among these severe malaria cases. P. falciparum submicroscopic infection may lead to severe malaria in perennial transmission area.

基于指纹图谱和化学模式识别评价杜仲叶质量

目的建立杜仲Eucommia ulmoides叶高效液相色谱(HPLC)指纹图谱,结合化学模式识别评价其质量。方法采用Ultimate XB-C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%磷酸溶液为流动相梯度洗脱,体积流量1.0 mL/min,柱温30℃,进样量20μL,波长为207、240、277、350 nm,检测杜仲叶中12种活性成分含量并建立指纹图谱,应用SPSS 25.0和SIMCA 14.1软件进行聚类分析(clustering analysis,CA)、主成分分析(principal component analysis,PCA)和偏最小二乘法-判别分析(partial least squares discrimination analysis,PLS-DA),对不同产地杜仲叶进行质量评价。结果不同产地杜仲叶桃叶珊瑚苷、京尼平苷酸、原儿茶素、绿原酸、儿茶素、车叶草苷、咖啡酸、京尼平苷、松脂醇二葡萄糖苷、芦丁、异槲皮素、紫云英苷差异显著,质量分数范围分别为0.053%~4.563%、0.078%~0.977%、0.018%~0.084%、2.634%~6.183%、0.010%~0.088%、0.124%~0.582%、0.001%~0.242%、0.033%~0.163%、0.065%~0.215%、0.045%~0.865%、0.199%~0.910%、0.020%~0.221%;20批杜仲叶HPLC指纹图谱确定了25个共有峰,经对照品指认11个色谱峰,相似度0.905~0.996;通过聚类分析将样品按不同产地分为2大类,最后筛选出绿原酸、桃叶珊瑚苷、车叶草苷、芦丁、松脂醇二葡萄糖苷是导致不同产地样品差异的标志性化合物。结论建立的指纹图谱方法稳定、可靠、重复性好,结合化学模式识别可用于杜仲叶质量评价。

Early diagnosis and monitoring of active HCMV infection in children with systemic lupus erythematosus

Objective To investigate the prevalence and features of active human cytomegalovirus (HCMV) infection in children with systemic lupus erythematosus (SLE) and evaluate the diagnostic value of the HCMV using antigenemia assay, serum polymerase chain reaction (PCR) and serology test. Methods Twenty-one SLE children undergoing immunosuppressive therapy were enrolled in this study. Immunofluorescence assay, PCR and serology tests were used to determine HCMV pp65 and p72 antigens in leukocytes, HCMV DNA in sera, and HCMV specific IgM and IgG antibodies, respectively. As a control group, twenty-one immunocompetent children with skeletal malformation were involved in this study. Statistical analysis was performed using Chi-square test or Fisher's exact test (Systat, USA), P values less than 0.05 were considered significant.Results Active HCMV infection was diagnosed in 28.6% (6/21) of SLE patients, with none in the control group; the difference between the two groups was significant (P=0.027). Two out of 6 SLE patients developed active HCMV infection before immunosuppressive therapy and the remaining 4 patients developed SLE after immunosuppressive therapy. Among the 21 SLE children, HCMV pp65 antigenemia was detected in 5 patients, p72 antigenemia in 3 patients, serum HCMV DNA in 9 patients, serum HCMV-specific IgM in 2 patients, and IgG in 19 patients. The sensitivity and specificity for diagnosis of active HCMV infection were 83.3% and 100%, respectively for pp65 antigenemia; 50% and 100% for p72 antigenemia; 100% and 80% for serum PCR; 33.3% and 100% for HCMV IgM serology; 50% and 100% for HCMV IgG serology. Conclusions Compared with the control group, active HCMV infection is much more frequent in SLE children, and can occur before treatment with immunosuppressive agents, but most often occur after immunosuppressive therapy. In comparison with the other techniques used in this study, the pp65 antigenemia assay seems to be a better method for the early diagnosis and monitoring of active HCMV infection in children with SLE.