To establish a new method for identifying genus of Lilium by DNA barcoding technology, ITS, ITS2, psbA-trnH, matK and rbcL sequences were analyzed in term of variation of inter- and intra-species, barcoding gap, neighbor-joining tree to distinguish genus of Lilium based on 978 sequences from experimental and GenBank database, and identification efficiency was evaluated by Nearest distance and BLAST1 methods. The results showed that DNA barcoding could identify different species in genus of Lilium. ITS sequence performed higher identification efficiency, and had significant difference between intra- and inter-species. And NJ tree could also divide species into different clades. Results indicate that DNA barcoding can identify genus of Lilium accurately. ITS sequence can be the optimal barcode to identify species of Lilium.
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Lilium ; classification

This study provides the candidate sequences in the identification of Radix et Rhizoma Clematidis and its adulterants using DNA barcoding. We amplified and sequenced the region psbA-trnH, with the data of 284 sequences from GenBank, the differential intra- and inter-specific divergences, genetic distance, barcoding gap were used to evaluate five barcodes, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The results showed that psbA-trnH barcodes performed high identification efficiency and inter-specific divergences among the five different DNA barcodes. Analysis of the barcoding gap and NJ tree showed psbA-trnH was superior to other barcodes. Based on the identification and PCR amplification efficiency, psbA-trnH can be the ideal barcode to identify Radix et Rhizoma Clematidis and its adulterants accurately.
DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; Drug Contamination ; Nucleic Acid Amplification Techniques ; methods ; Plant Roots ; genetics ; Plants, Medicinal ; classification ; genetics ; Ranunculaceae ; classification ; genetics ; Rhizome ; genetics ; Species Specificity

Antineoplastic Agents ; adverse effects ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Neoplasms ; drug therapy ; Ondansetron ; therapeutic use ; Phytotherapy ; Vomiting, Anticipatory ; drug therapy

HPLC analysis was performed to study the changes in chemical composition of ginseng extracts prepared from high quality ginseng with 0, 2, 4, 8 h of steamed times. An UFLC-MS/MS multiple-reaction monitoring (MRM) quantitative analysis was made to investigate the pharmacokinetic behavior differences of ginsenosides in mice ig administered of ginseng extracts with different steamed times in the negative ion mode, with Digoxin as the internal standard substance. The mice were injected with LPS to establish inflammation model after ig administration of ginseng for a week and the contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in mice plasma were detected by ELISA, in order to study on anti-inflammatory effects of ginseng with different steamed times. It was determined that levels of TNF-α and IL-1β were significantly decreased in inflammation model group ig administered of ginseng extracts with 8h of steamed time. The results showed that the chemical components in ginseng changed after steaming and the components into the blood changed, correspondingly. Ginseng with steamed 8 h contributes to anti-inflammatory effects. These results provided an experimental basis for revealing the active substance basis and dose-effect relationship of ginseng on anti-inflammatory effect.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacokinetics ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Ginsenosides ; chemistry ; pharmacokinetics ; Hot Temperature ; Humans ; Inflammation ; drug therapy ; Male ; Mice ; Mice, Inbred ICR ; Panax ; chemistry ; Time Factors

Objectives To investigate the characteristics of changes in serum metabolic profile before and after resection of carcinoma tissues to establish a disease distinguishing model,to analyze the changing trend of characteristic metabolites,and to determine the molecular mechanism and potential clinical value of characteristic metabolic markers for HBV-related liver cancer.Methods Ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) was used to analyze the serum metabolites of 15 patients with hepatocellular carcinoma (HCC) before and after partial hepatectomy and on 25 healthy volunteers.The pattern recognition method and nonparametric test analyzes were used to analyze the data and to identify the specific metabolites and their changes after resection of carcinoma tissues.Results We established the principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) disease distinguishing model for HCC patients before and after operation as against the healthy volunteers.To distinguish between the liver cancer group and the normal control group,27 characteristic metabolites were selected from the patients before and after resection of carcinoma tissues.Eight moved towards the normal control after resection of carcinoma tissues.This indicated that liver carcinoma was an important impacting factor for these metabolites.Finally,7 metabolites were identified,and these metabolites had high diagnostic value as shown on ROC curves.Conclusions Through serum metabolic profiling of patients before and after resection of carcinoma tissues,a high correlation between metabolism and hepatocellular carcinoma was found.Researches on endogenous metabolites and pathways in liver diseases will provide a better understanding of the molecular mechanisms and provide further directions for clinical diagnosis and treatment.

Objective To evaluate the role of Ras homolog family member A (RhoA)/Rho-associated coiled-coil containing protein kinase 2 (ROCK2) signaling pathway in propofol-induced neuroapoptosis in the hippocampus of newborn rats.Methods Experiment Ⅰ Primarily cultured hippocampal neurons were seeded in 6-well culture plates at a density of 1×106 cells/ml and divided into 2 groups (n=6 each) using a random number table:solvent control group (C group) and propofol group (P group).Propofol was added with the final concentration of 60 μg/ml in group P.Dimethyl sulfoxide was added with the final concentration of 0.04％ in group C.The expression of RhoA and ROCK2 in hippocampal neurons was measured by Western blot at 24 h of incubation.Experiment Ⅱ Primarily cultured hippocampal neurons were seeded in 6-well culture plates at a density of 1 × 106 cells/ml and divided into 3 groups (n =6 each) using a random number table:solvent control group (C group),propofol group (P group) and propofol plus specific RhoA/ROCK2 signaling pathway blocker Y27632 group (P+Y group).Propofol was added with the final concentration of 60 μg/ml in group P.Propofol at the final concentration of 60 μg/ml and Y27632 at the final concentration of 10 μmol/L were added in group P+Y.Dimethyl sulfoxide was added with the final concentrauon of 0.04％ in group C.At 24 h of incubation,the neuroapoptosis in hippocampi was detected by flow cytometry,and the expression of activated caspase-3 in hippocampal neurons was measured by Western blot.The apoptotic rate was calculated.Results Experiment Ⅰ Compared with group C,the expression of RhoA and ROCK2 in hippocampal neurons was significantly up-regulated in group P (P＜0.05).Experiment Ⅱ Compared with group C,the apoptotic rate of hippocampal neurons was significantly increased,and the expression of activated caspase-3 was up-regulated in P and P+Y groups (P＜0.05 or 0.01).Compared with group P,the apoptotic rate of hippocampal neurons was significantly decreased,and the expression of activated caspase-3 was down-rcgulatcd in group P + Y (P＜ 0.05).Conclusion Activation of RhoA/ROCK2 signaling pathway is involved in propofol-induced neuroapoptosis in hippocampi of newborn rats.

OBJECTIVETo explore the effect of Telbivudine (LDT) Tablet combined with Jianpi Bushen Recipe (JBR) on serum hepatitis B virus (HBV) specific cytotoxic T lymphocyte (CTL) and HBeAg seroconversion in chronic hepatitis B (CHB) patients.

METHODSTotally 90 HBeAg-positive and human leukocyte antigen (HLA)-A2 positive CHB patients were randomly assigned to the treatment group and the control group, 45 cases in each group. Patients in the treatment group took LDT Tablet (600 mg, once per day) combined with JBR granule (twice per day), while those in the control group took LDT Tablet alone. The therapeutic course for all was one year. HBV DNA negative conversion rate, HBeAg seroconversion rate, and level of HBV specific CTL were compared after 1 year treatment; liver function, drug resistance mutations, and adverse reactions were also compared between the two groups.

RESULTSAfter 1 year treatment, HBV DNA negative conversion rate and HBeAg seroconversion rate were 88.89% (40/45) and 40.00% (18/45) in the treatment group, higher than those of the control group [68.89% (31/45) and 20.00% (9/45)], with statistical difference (P < 0.05). Level of HBV specific CTL in the treatment group was 0.78% +/- 0.09% after treatment, higher than that of the control group after 1 year treatment (0.54% +/- 0.11%) and that before treatment (0.36% +/- 0.07%), with statistical difference (P < 0.01). Level of HBV specific CTL in 27 patients with HBeAg seroconversion was 0.81% 0.10%, higher than that of 63 patients without HBeAg seroconversion (0.60% +/- 0.09%), with statistical difference (P < 0.01). ALT returned to normal in 44 cases of the treatment group (97.78%), while it was 42 cases (93.33%) of the control group, with no statistical difference between the two groups (P > 0.05). Total bilirubin (TBil) in the two groups all turned to normal. rtM204I variation occurred in 1 case (2.22%) of the treatment group and 2 cases (4.44%) in the control group. No obvious adverse reaction occurred in the two groups.

CONCLUSIONLDT Tablet combined with JBR could elevate levels of HBV specific CTL and HBeAg seroconversion in CHB patients.

Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; Hepatitis B, Chronic ; drug therapy ; Humans ; Seroconversion ; T-Lymphocytes, Cytotoxic ; immunology ; Tablets ; Thymidine ; analogs & derivatives ; therapeutic use

Rat aorta media, adventitia and cultured vascular smooth muscle cells (VSMCs) were used in this study to identify the source of nitric oxide (NO) generation from various cell types of vascular tissues and to elucidate the mechanisms involved in NO formation. Treatment of vascular media and VSMCs with lipopolysaccharide (LPS) or cytokines [tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta)] resulted in a dose-dependent increase of NO release. Inducible nitric oxide synthase (iNOS) in the stimulated VSMCs was significantly upregulated as shown by Western blot analysis. Protein kinase C (PKC) inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7) prevented LPS-, TNF-alpha- and IL-1beta-induced NO production, whereas N-(2-guanidinoethyl)-5-isoquinoline-sulfonamide (HA1004), an H7 analogue with little activity towards PKC, had no inhibition effect. The role of PKC in LPS- and cytokine-induced changes on NO formation was confirmed by using another structurally distinct PKC inhibitor chelerythrine. Treatment of VSMCs with protein tyrosine kinase (PTK) inhibitor genistein or tyrphostin AG18 also reduced the NO production evoked by LPS, TNF-alpha or IL-1beta, which was associated with inhibition of iNOS protein expression. In contrast, PKC inhibitor chelerythrine did not affect iNOS expression. These results suggest that PTK mediates LPS- and cytokine-induced NO formation by upregulation of iNOS expression. PKC may be involved in the post-translational modification of iNOS or the regulation of the availability of iNOS substrates and cofactors.
Animals ; Aorta, Thoracic ; cytology ; Cells, Cultured ; Cytokines ; pharmacology ; Interleukin-1beta ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Protein Kinase C ; physiology ; Protein-Tyrosine Kinases ; physiology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; pharmacology

Amino Acids ; analysis ; Animals ; Chromatography, Gas ; methods ; Drug Contamination ; Electrophoresis, Polyacrylamide Gel ; Materia Medica ; chemistry ; Quality Control ; Swallows ; classification

Objective To investigate the serum levels of endothelin-1 (ET-1), tumor necrosis factor- α (TNF-α) and their dynamic changes in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) and their relation with the condition variation of the patients.Methods The serum ET-1 and TNF-α levels were determined with enzyme-linked immunno-sorbent assay (ELISA) in 31 patients with DEACMP, and the dynamic changes of patients' condition were monitored by use of the activity of daily living (ADL) scale, the information-memory-concentration test (IMCT) and the Hasegawa's dementia scale (HDS). The comparisons between patients with DEACMP and both 30 patients with acute carbon momoxide poisoning (ACMP) but without DEACMP and 30 normal controls were also conducted. Results At the acute stage of the DEACMP group, the serum levels of ET-1 and TNF-α were both significantly higher than those in the normal control group (P＜0.05); that of TNF-α was significantly lower than that in the ACMP group (P＜0.05), but that of ET-1 was not significantly different from that in the ACMP group (P＞0.05). The serum levels of ET-1 and TNF- α in the ACMP group were both significantly higher than those in the normal control group(P＜0.05). In the DEACMP group, the serum level of ET-1 at the convalescent stage was significantly lower than that at the acute stage (P＜0.05), but the serum level of TNF-α was not significantly different from that at the acute stage (P＞0.05). At the acute stage of the DEACMP group, ADL scores were significantly higher than those in norms, and IMCT scores and HDS scores were significantly lower than those in norms (P＜0.05). In the DEACMP group, the ADL scores at the convalescent stage were significantly lower than those at the acute stage (P＜0.05), IMCT scores and HDS scores were significantly higher than those at the acute stage (P＜0.05). Significant correlations between scores of any 2 of 3 scales in patients with DEACMP at both acute and convalescent stage were noted (P＜0.05). Conclusion The dynamic detection of serum ET-1 and TNF-α level variations could be used as an indicator for condition severity in patients with DEACMP.