Simultaneous Determination of Baicalin, Berberine and Rhein by HPLC in Traditional Chinese Patent Medicine Sanhuang Tablets

Aim To establish a reversed phase liquid chromatographic method forsimultaneous determination of three main medicinal constituents, baicalin, berberine and rhein, inSanhuang tablets. Methods The separation was performed on a Kromasil C_(18) column with TEA-adjusted0.02 mol·L^(-1) H_3PO_4 (pH 6.78)-acetonitrile-methanol (40 : 9 : 7) as mobile phase at aflow-rate of 1.0 mL·min^(-1), with detection at 254 ran. Considering interaction between acidic andalkaline compounds, three standard markers were added respectively and the volume of samplesolution was doubled in recovery experiments. Results Three regression equations revealed excellentlinear relationship between the peak areas and concentrations and the correlation coefficients allsurpassed 0.999 8. The average recovery was 96.1% (RSD = 2.1%) baicalin, 98.5% (RSD = 2.4%) forberberine, and 101.5% (RSD =1.3%) for rhein. Conclusion The method developed can be used to controlthe quality of Sanhuang tablets comprehensively.

Construction of Fusion Expression Vector of α-galactosidase-EGFP in Cucumber

[Objective] The research aimed to construct the fusion protein expression vector of α-galactosidase-EGFP （enhanced green fluorescent protein） in cucumber controlled by CaMV35S promoter.[Method] CaMV35S promoter sequence and the coding region of EGFP were amplified by polymerase chain reactions （PCR） with vector pCambia 1303 as the template.Using reverse transcript PCR technology,with total RNAs of cucumber as template,the coding region of acid α-galactosidase Ⅰ in cucumber was amplified.The above three fragments were inserted into the multiple cloning sites of expression vector pCambia 1381c.The fusion expression vector of α-galactosidase-EGFP located at the C-terminal of the target genes was constructed.[Result] After enzyme digestion and sequencing,the fusion expression of α-galactosidase-EGFP in cucumber was constructed successfully.[Conclusion] The research laid the experimental basis for further study on the subcellular localization of α-galactosidase in cucumber.

南朝鲜发酵工程产品信息

1.发酵法生产5′-IMP:菌体系产氨短杆菌突变株,对Mn++不敏感,腺嘌呤、鸟嘌呤双重缺陷型。产酸率40～50[克/升]。 2.发酵法生产5′-XMP（黄苷酸）,酶法转化成5′-GMP,发酵5′-XMP产率60～70（克/升）,经产氨短杆菌酶法转化为5′-GMP,转化率在90％以上。 3.RNA酶法水解生产5′-AMP、5′-CMP、5′-UMP。

ALCA-R可以促进ddI的代谢以及抗HIV的功能

2′,3′一双脱氧次黄苷(ddI)的抗病毒活性取决于其在细胞内转化为三磷酸双脱氧次黄苷((ddITP)。5—氨基—4咪唑羧酰胺(ALCA-R)是嘌呤生物合成过程中一天然代谢产物,它可转化为次黄苷酸(IMP)。后者是腺苷和鸟苷生物合成的底物,因此可以增进细胞内嘌呤核苷酸的代谢库。同时,在由细胞浆5,—核苷酸酶催化 ddI 磷酸化并产生单磷酸双脱氧次黄苷的过程中,IMP 可以提供磷酸基。因此,在本文中,我们研究 ALCA-R 对ddI 磷酸化以及抗病毒活性的影响。

Characteristics of Deposition of Inosine Monophosphate(IMP) and Intramuscular Fat(IMF) in Muscles of Jinghai Yellow Chicken and Its Crossbreeds

[Objective] This study aimed to investigate the characteristics of deposition of inosine monophosphate （IMP） and intramuscular fat （IMF） in muscles of Jinghai yellow chickens and its crossbred.[Method] The characteristics of IMP and IMF deposition of 112-day-old Jinghai yellow chickens （J×J） and its two different 70-day- old crossbreeds （J×B and B×B） were analyzed. The IMP content in breast muscle and leg muscle were determined by HPLC. [Result] The contents of IMP and cor- rected inosine monophosphate （IMPc） in breast muscle were significantly or ex- tremely significantly higher than that in leg muscle of the chickens in the three groups whether in male or female chickens （P〈0.05 or P〈0.01）. There were no sig- nificant difference in the contents of IMP and IMPc between hens and roosters （P〉 0.05）. The fresh degree of breast muscle and leg muscle was 96,11%-98.16% and 87.22%-93.07%, respectively. And the fresh degree of breast muscle was higher than that of leg muscle. In the three groups, the IMF content in leg muscle was significantly higher than that in breast muscle whether in male or female chickens （P〈0.05）. The contents of IMF in breast muscle and leg muscle were 0.36%-0.75% and 1.84%-2.38%, respectively. The iMP content in breast muscle of chickens in Bx J group was extremely significantly higher than that in breast muscle of chickens in JxJ group （P〈0.01）, but the contents of IMPc and iMF of breast muscle and leg muscle of the chickens in the three groups had no significant difference （P〉0.05）. [Conclusion] To sum up, the freshness and flavor significantly differ between the breast muscle and leg muscle of chickens, but show no significant difference among the three groups.

Population genetic structure of the mantis shrimp Oratosquilla oratoria(Crustacea:Squillidae)in the Yellow Sea and East China Sea

The mantis shrimp O ratosquilla oratoria is an ecologically and economically important species in the Western Pacific. In present study, the population genetic structure of O ratosquilla oratoria from the Yellow Sea and East China Sea was examined with mitochondrial DNA control region sequences. In total, 394 samples were collected from 18 locations and 102 haplotypes were obtained. For the Yellow Sea, the overall nucleotide diversity and haplotype diversity were 0.006 9 and 0.946 8, respectively; while across all the East China Sea locations, the overall nucleotide diversity and haplotype diversity were 0.027 94 and 0.979 0, respectively. The results of AMOVA and pairwise F_(ST)(0.145 2, P <0.001) revealed moderate differentiation between the Yellow Sea and East China Sea populations of O. oratoria. However, neither the neighbor-joining tree nor haplotype network showed clades with geographic pattern, which indicated considerable gene flow was existed between the Yellow Sea and East China Sea, and supporting the high larval dispersal ability in this species. Mismatch distribution analysis and neutrality tests suggested that O. oratoria has undergone population expansion event, and the Pleistocene glacial cycles might have an impact on the historical demography of O. oratoria. The genetic information obtained in this study can provide useful information for sustainable improvements for capture fisheries management strategies.

卡介苗联合Poly I：C接种对新生期小鼠脾脏T细胞的影响

目的研究卡介苗（BCG）联合Poly I：C新生期接种对小鼠脾脏T细胞功能亚群发育的影响。方法新生清洁级BALB／c小鼠32只，分4组：对照组、BCG组、Poly I：C组、BCG和Poly I：C联合接种组，新生期接种4周后取脾细胞用流式细胞仪检测CD3^＋CD8^＋IFN-γ^＋、CD3^＋CD8^-IFN-γ^＋、CD3^＋CD8^＋IL-4^＋、CD3^＋CD8^-IL-4^＋、CD4^＋Foxp3^＋T细胞的比例，其分别代表Te1、TH1、Tc2、TH2和调节性T细胞（Treg）亚群。结果BCG组、Poly I：C组、BCG和Poly I：C联合接种组TH1、Tc1细胞比例明显高于对照组（P〈0．05或P〈0．01），3个接种组间比较差异无统计学意义；Tc2、TH2和Treh比例各组间比较差异没有统计学意义；3个接种组TH1／TH2和总IFN-γ／IL-4比值明显高于对照组（P〈0．05或P〈0．01），联合接种组TH1／TH2比值高于BCG组（P〈0．05），Tc1／Tc2比值各组间差异没有统计学意义；CD4^＋Foxp3^＋T细胞比例各组间比较差异没有统计学意义。结论新生期BCG和Poly I：C接种均能促进TH1、Te1细胞亚群的发育，提高TH1／TH2比值，其联合接种在TH1／TH2水平可能具有一定的协同作用，但不能影响CD4^＋Foxp3^＋Treg细胞的比例。

异源蓝舌病毒dsRNA与PolyI：C诱导人源体细胞产生IFN-β的比较研究

目的探讨异源动物蓝舌病毒dsRNA诱导人源细胞产生干扰素（IFN）的能力。方法借助体外培养系统，利用ELISA技术检测IFN诱导剂多聚次黄苷酸-胞苷酸（PolyI：C）、蓝舌病毒（BTV）及BTV dsRNA3种不同处理因素作用后，培养细胞上清液中IFN-β的含量，以比较研究其诱导人肺癌细胞A549和人胚肺细胞HEL产生IFN-β的能力和特征。结果这3种因子均能诱导人肺癌A549细胞和人正常肺细胞HEL产生IFN-β，BTVdsRNA诱导IFN-β产生的能力显著高于PolyI：C和BTV。浓度高的BTVdsRNA诱导细胞产生的IFN-β水平高，说明二者之间存在剂量依赖性。BTVdsRNA对不同性质的人源细胞（人肺癌A549细胞、人正常肺细胞HEL）具有不同的诱导产生IFN-β的能力，诱导人正常肺细胞HEL产生IFN-β的能力显著地高于诱导人肺癌细胞A549产生IFN-β的能力（P〈0．05）。结论裸露的BTVdsRNA分子能有效地诱导人源细胞产生IFN-β；结合BTV和dsRNA对人的安全性以及人类正在努力寻找新的内源性IFN诱导剂的事实，可以推断BTV dsRNA具有发展成内源性IFN诱导剂之潜能。

Isolation and structural elucidation of an unstable isoflavonoid glycoside malonate from Radix hedysari

Based on the stability results of Radix hedysari extract, an unstable compound was extracted using acidic methanol and purified by rapid chromatographic methods successfully from Radix hedysari for the first time guided by HPLC analysis. Its structure was identified as formononetin 7-O-β-D-（6＂-O-malonyl）-glucopyranoside by spectroscopic analyses.

Metabolite identification and the development of a simultaneous quantification method for wogonin and wogonin-7-O-glucuronide:application to a distribution study in mice liver

Wogonin possesses potent inhibitory activities against cancer cell growth in vitro and in vivo and has attractive safety profiles.A highly sensitive liquid chromatography coupled with tandem mass spectrometry method was developed for the identification of major metabolites in mice liver after intravenous administration of wogonin.Five metabolites were identified and biotransformation pathways were elucidated as well.Furthermore,a method was developed and validated for the simultaneous quantitatively determination of wogonin and wogonin-7-O-glucuronide in mice liver.After liquid-liquid extraction by ethyl acetate,the analytes were separated on a C_（18）column with a mobile phase of methanol-10 mM ammonium acetate water （80：20,v/v）.The detection was operated with negative selected reaction monitoring mode using electrospray ionization technique. The linear response range was 0.2-40μg/g for both wogonin and wogonin-7-O-glucuronide in mice liver.The developed quantification method was suitable for distribution study after intravenous infusion of wogonin injection in animals.

Simultaneous determination of 6 organic acids,3 nucleosides,and ephedrine in Pinellia ternata by HPLC

Pinellia ternata has a long history of medicinal use in China, Japan, and Korea. Herein, we present a sensitive, selective, and robust HPLC method for the simultaneous determination of 6 organic acids （oxalic acid, malic acid, citric acid, formic acid, acetic acid, and succinic acid）, 3 nucleosides （uridine, guanosine hydrate, and adenosine）, and ephedrine in P. ternata. The Rhizoma pinelliae samples were collected from different parts of Southwest China, and the 10 constituents were detected at a wavelength of 210 nm. Chromatographic separation was achieved on a Boston Green ODS reversed-phase column along with a gradient mobile phase of methanol and phosphate buffer （pH 2.3, 10 mmol/L, v/v） at a flow rate of 1.0 mL/min. The range of recovery was between 90.8% and 101.9%, and the calibration curves showed high linearity. The relative standard deviations of the new method were less than 3.0% for intra- and inter-day analyses. All samples were stable for at least 48 h. The abundance of the 10 compounds were dependent on their specific locations. The new HPLC method is a powerful technique for the quality control ofP. Ternata.

Bovine HSD17B8 gene and its relationship with growth and meat quality traits

17 β-Hydroxysteroid dehydrogenase type 8 （HSD17B8） is an important regulator of lipid and steroid metabolism. In the present study, we aimed to assess the effects of HSD17B8 on growth and meat quality traits in cattle. Transcription profile analysis showed that HSD17B8 was primarily expressed in the salpinx, liver, and testis. Meanwhile, we identified three SNPs （SNPI： intron 1-A91G; SNP2： exon 1-A90G; and SNP3： intron 8-A86G） of the bovine HSD17B8 gene and investigated its haplotype frequencies and linkage disequilibrium. The detected SNPs were found associated with growth traits （body weight, body length, height at withers, heart girth, hip width, and average daily gain） in native cattle populations （Nanyang and Jiaxian） as well as the meat quality traits （Warner- Bratzler shear force, rib area, dressing percentage, carcass weight, and backfat thickness） in commercial breeds （Angus, Hereford, Limousin, Luxi, Simmental, and Jinnan）. Our results provided evidence that polymorphisms in the HSD17B8 gene were associated with growth traits and meat quality traits. Moreover, our findings might be used for marker-assisted selection in beef cattle breeding program