Current quality control patterns are limited to industrial application, for most of the natural chemical reference substances are expensive and unavailable. Herein, a method, quantitative analysis of multi-components with single marker (QAMS), was established and validated to simultaneously determine nine ginsenosides (ginsenoside Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2, Rb3, Rd) in P. ginseng and four ginsenosides (ginsenoside Rg1, Rh1, Rb1, Rd) in P. notoginseng. Using ginsenoside Rb1 as the contrast, the relative correction factors (RCF) of the other eight ginsenosides were determined by HPLC-DAD. Within the linear ranges, the values of RCF of ginsenoside Rb1 to ginsenoside Rg1, Re, Rf, Rh1, Rc, Rb2, Rb3 and Rd were 1.400, 1.215, 1.517, 1.801, 0.944, 1.012, 1.143, and 1.135, respectively. The RCF had a good reproducibility in various instruments, chromatographic columns (RSD = 0.30% - 3.9%). According to their RCF, we simultaneously determined nine ginsenosides in P. ginseng only using one marker. In addition, the RCF of ginsenosides were used to simultaneously quantitative analysis of four ginsenosides in P. notoginseng. The results of QAMS method were validated by comparing with that of external standard method, and no obvious significant difference was found.
Chromatography, High Pressure Liquid ; methods ; Ginsenosides ; analysis ; Panax ; chemistry ; classification ; Quality Control ; Reproducibility of Results

OBJECTIVETo study and research the transcription pattern of UL131A-128 mRNA in human cytomegalovirus (HCMV) clinical low passage strains.

METHODSThe UL131A-128 mRNAs of from different clinical strains and kinetic periods were amplified using 3' RACE and analyzed by sequencing. Meanwhile, clones containing UL131A-128 transcripts in a HCMV cDNA library of clinical strain were selected and sequenced.

RESULTSIt is successful to obtain the transcription pattern of UL131A, UL130 and UL128 gene in HCMV clinical low passage strains, the UL131A gene consisted of two exons and the coding region of UL130 gene was not interrupted by any intron in the region as reported before. However, the transcript of UL128 gene showed two patterns, one pattern consisted of the three exons, the length is 519bp, and the other one contained the three exons and the sequence of the first intron further, the length is 642bp. The quantities of UL128 transcript containing the sequence of the first intron were higher than that of transcript only containing the three exons in the studied clinical strains at all kinetic classes. It was demonstrated that the UL131A-128 mRNA were expressed with immediately early, early and late kinetics. The result of 3'RACE and HCMV cDNA library of clinical strain is conformity.

CONCLUSIONSOur results demonstrated that the UL131A, UL130 and UL128 genes were transcribed with 3'-coterminal, although the initiation points of their mRNA may be different. The variation of the transcripts found in our study indicated complex nature of transcription of UL131A-128 genes in HCMV clinical strains.

Cytomegalovirus ; genetics ; Gene Library ; Humans ; Membrane Glycoproteins ; genetics ; RNA, Messenger ; analysis ; Transcription, Genetic ; Viral Envelope Proteins ; genetics

OBJECTIVETo observe the performance of liver metastases with contrast-enhanced ultrasound (CEUS) and assess its clinical application.

METHODSTwenty-one patients with 21 untreated liver metastases underwent CEUS with low mechanical index imaging. The characteristic appearances of CEUS in different vascular phases were observed.

RESULTSOf 21 metastases, 19 (90.5%) were identified as fast-in and fast-out enhancement pattern. In the arterial phase, all the 21 lesions showed enhancement but with varied appearances: 12 (57.1%) showed early diffuse enhancement, 8 (38.1%) showed ring-like enhancement, and the remaining one lesion of large size showed slowly enhomogenous minor enhancement. In the late phase, sharp defects were found in 20 lesions (95.2%), and more lesions were detected in 3 patients (14.3%).

CONCLUSIONCEUS can show the characteristic appearance of liver metastases, and can be used for the screening and diagnosis of liver cancers.

Contrast Media ; Humans ; Liver Neoplasms ; diagnostic imaging ; secondary ; Ultrasonography

The regulator of expression of virion(Rev)protein binds specifically to the Rev-responsive element(RRE)RNA in order to regulate the expression of the human immunodeficiency virus(HIV)-1 genes.Fluores-cence indicator displacement assays have been used to identify ligands that can inhibit the Rev-RRE interaction;however,the small fluorescence indicators cannot fully replace the Rev peptide or protein.As a result,a single rhodamine B labeled Rev(RB-Rev)model peptide was utilized in this study to develop a direct and efficient Rev-RRE inhibitor screening model.Due to photon-induced electron transfer quenching of the tryptophan residue on the RB fluorophore,the fluorescence of RB in Rev was weakened and could be dramatically reactivated by interaction with RRE RNA in ammonium acetate buffer(approximately six times).The interaction could reduce the electron transfer between tryptophan and RB,and RRE could also increase RB fluorescence.The inhibitor screening model was evaluated using three known positive Rev-RRE inhibitors,namely,proflavin,6-chloro-9-[3-(2-chloroethylamino)pro-pylamino]-2-methoxyacridine(ICR 191),and neomycin,as well as a negative drug,arginine.With the addition of the positive drugs,the fluorescence of the Rev-RRE decreased,indicating the displacement of RB-Rev.This was confirmed using atomic force microscopy(AFM)and the fluorescence was essentially unaffected by the addition of arginine.The results demonstrated that RB-Rev can be used as a fluorescent probe for recognizing small ligands that target RRE RNA.The Rev-RRE inhibitor screening model offers a novel approach to evaluating and identifying long-acting Rev inhibitors.

To explore the possible mechanism of liver injury, the effects of Ploygoni Multiflori Caulis and its extractive on the function of bilirubin-associated transporters were investigated in normal (N) and idiosyncratic (LPS) rats (M). The normal and LPS rats were respectively administrated powder of Ploygoni Multiflori Caulis, its extractive and same volume of 0.5% CMC-Na solution for 7 d. BSP, a substrate of the transporters of Oatp1a1 and Oatp1b2 was selected, and its pharmacokinetic parameters of intravenous injection were determined to examined the activity these transporters. Meanwhile the mRNA expressions of transporters were detected. Compared with N-blank control group, besides M-powder group, the Cmax has no significantly different from other groups, t1/2, AUC0-t and AUC0-∞ were significantly increased, and CL were significantly decreased. However, compared with N- blank control group, AST and ALT decreased significantly. The expression of Oatp1a1, Oatp1b2 and MRP2 mRNA was significantly decreased (P<0.05), but there was no act synergistically when Ploygoni Multiflori Caulis and extractive were combined with LPS. The function of Oatp1a1, Oatp1b2 and MRP2 in rats were significantly inhibited by Ploygoni Multiflori Caulis and extractive, which may be an important cause of hepatotoxicity.

OBJECTIVE: To investigate the changes in serum levels of endothelin-1 (ET-1) and connective tissue growth factor (CTGF) in patients with atrial fibrillation (AF) and their value for predicting recurrence of AF after radiofrequency ablation (RFCA). METHODS: Sixty-six patients with paroxysmal AF (PaAF) and 72 with persistent AF (PaAF) admitted in our hospital were recruited as AF group and 80 patients with sinus rhythm as the control group, and in all the participants, serum levels of ET-1 and CTGF were measured using ELISA and Western blotting. From 6 patients with AF and 6 with sinus rhythm undergoing cardiac surgery in our hospital, tissue samples of the right atrial appendage were taken intraoperatively for observation of structural changes of the cardiomyocytes, myocardial fibrosis and expression of ET-1 and CTGF protein. In AF group, the patients receiving RFCA were followed up for 6 months following the procedure for assessment of the outcomes. RESULTS: Compared with the control patients, the patients with AF showed obvious damages of the cardiomyocyte structure and myocardial fibrosis. Serum levels of ET-1 and CTGF levels were significantly higher in PaAF and PeAF groups than in the control group, and were higher in PeAF group than in PaAF group. In the patients with AF, serum ET-1 and CTGF levels were positively correlated with left atrial diameter (LAD) (P < 0.05), and ET-1 was positively correlated with CTGF levels (P < 0.05). In patients with postoperative AF recurrence, the serum levels of ET-1 and CTGF were significantly higher than those in patients without recurrence; serum ET-1 and CTGF levels before and after the operation were positively correlated with the recurrence of PeAF, and elevated serum levels of ET- 1 and CTGF were identified by logistic regression analysis as independent risk factors for postoperative recurrence of PeAF. CONCLUSION Serum levels of ET-1 and CTGF are significantly elevated in AF patients in positive correlation with AF duration. ET-1 and CTGF levels are higher in AF patients with postoperative recurrence, and they both have predictive value for recurrence of PeAF following RFCA.
Humans ; Atrial Fibrillation ; Endothelin-1 ; Connective Tissue Growth Factor ; Chronic Disease ; Atrial Appendage ; Fibrosis

An eco-friendly and fast HPLC method was developed for the determination of adenosine, inosine, guanosine and uridine in Cordyceps and related products (fermented mycelia of Hirsutella sinensis andPaecilomyces hepiali). The sample was ultrasonically extracted using 0.5% phosphoric acid solutions for 2.5 min. Sample separation was performed on a Poroshell SB-Aq column (50 mm × 4.6 mm, 2.7 μm) using eco-friendly mobile phase consisting of formic acid and ammonium formate aqueous solution at a flow rate of 1.0 mL·min
Adenosine ; Chromatography, High Pressure Liquid ; Cordyceps ; Nucleosides