This study aims to investigate the relationship between odor and contents of the chemical compounds in Lonicera japonica, including chlorogenic acid, galuteolin and polyphenols. High performance liquid chromatography (HPLC) was applied to determine the contents of chlorogenic acid and galuteolin in L. japonica. The ponptent of polyphenols was determined by UV-Vis Spectrophotometry. Electronic nose was used to extract and measure the odor of L. japonica. Then SPSS 17.0 software was employed for data processing. There is a significant positive correlation between the comprehensive index value of aroma and the contents of chlorogenic acid and polyphenols. The regression equations have been established. However, the relationship between the comprehensive index value and the content of galuteolin is not obvious. This is proof that the odor of L. japonica has close connection with the chemical compounds. Therefore, this research offered a new method for initially determine or predict the content of the chemical composition in L. japonica,
Chlorogenic Acid ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Electronic Nose ; Lonicera ; chemistry ; Odorants ; analysis ; Polyphenols ; chemistry ; Smell

Objective To investigate the diagnosis value of homocysteine (Hcy) combined with carcinoembryonic antigen (CEA) and squamous cell carcinoma (SCC) antigen for esophageal cancer.Methods The serum levels of Hcy,CEA,CA199,CA724,CA50 and SCC antigen in 163 patients with esophageal cancer and 49 healthy people were measured.The diagnosis efficacy between the combination of Hcy,CEA and SCC antigen and the combination of CEA,CA199,CA724,CA50 and SCC antigen was compared.Results The levels of Hcy,CEA and SCC antigen were significantly raised in patients group as compared with the levels in control group (all P ＜ 0.05).The area under the ROC curve of Hcy was 0.722 (95 ％ CI 0.633-0.811),CEA was 0.619 (95 ％ CI 0.533-0.704) and SCC antigen was 0.685 (95 ％ CI 0.608-0.762),respectively.There were no significant differences among the three area under the ROC curve.The sensitivity,specificity and accuracy of the combination of CEA,CA199,CA724,CA50,SCC antigen were 69.9 ％,59.2 ％ and 67.5 ％,respectively,but the sensitivity,specificity and accuracy of the combination of Hcy,CEA,SCC antigen were 96.3 ％,69.4 ％ and 90.1％,respectively.There was significant difference between the combination of Hcy,CEA,SCC antigen and the combination of CEA,CA199,CA724,CA50,SCC antigen (P ＜ 0.05).Conclusion The combination of Hcy,CEA,SCC antigen is helpful to determine the diagnosis and efficacy of esophageal cancer.

Viral hepatitis was the most common infectious disease in china. But the diagnosis and treatment were varied because the viral hepatitis patients were hospitalized in different kinds of hospital such as infectious disease hospital, general hospital and Chinese medical hospital. It was necessary to know clinical characters and information of viral hepatitis patients in different hospitals. The general information, subtype distribution, prognosis, complication, medication and relations of onset with solar term from 41 180 viral hepatitis patients based on HIS data were analyzed. It was found that the age of patients between 18 to 59 years old was most; most patients were males. The national basic medical insurance was the most type of payment. The outcome of viral hepatitis in the youth and female were better than that in the old and male. Acute hepatitis was easer to restore than chronic hepatitis. Liver cirrhosis and hepatocellular carcinoma were the two most complications. The peak of onset was during summer solstice, slight heat and great heat. The most common Chinese medicine was Diammonium glycyrrhizinate and the most common western medicine was reduced glutathione. The combination of D. glycyrrhizinate with reduced glutathione, polyene phosphatidylcholine and thymosin was the main pattern. But It was not knew if the combination of western and Chinese medicine was the most effective therapy to protect liver function. It was necessary to take deeply research of the relationship between the combination therapy and their effectiveness.
Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Glutathione ; therapeutic use ; Glycyrrhizic Acid ; therapeutic use ; Hepatitis, Viral, Human ; drug therapy ; Hospitals ; Humans ; Male ; Middle Aged ; Young Adult

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling ; Panax ; genetics ; Saponins ; biosynthesis ; Sequence Analysis, RNA

AIMTo investigate the effects of Nrf2 (Nuclear-E2 related factor) on gamma-glutamylcysteine synthase (gamma-GCS) in lung of guinea pigs with bronchial asthma.

METHODS20 adult male guinea pigs were randomly divided into two groups (n = 10): control group (C group) and asthmatic group (A group), asthmatic model was established by ovalbumin intraperitoneal and ovalbumin inhalation. The reactive oxygen piece (ROS), reduced glutathione (GSH), oxidant glutathione (GSSG) and total GSH in lung tissue were examined respectively. Inflammatory cell infiltration and index of remodeling of bronchiole were detected. In situ hybridization detected the gamma-GCS heavy subunit (gamma-GCS h) mRNA in lung tissue. Immunohistochemistry detected the expression of Nrf2 protein and gamma-GCS protein in lung tissue. RT-PCR measured the expression of Nrf2 mRNA in lung tissue. The activity of gamma-GCS was measured by coupled enzyme assay.

RESULTS(1) The number of eosinophils and lymphocytes in bronchiole of A group were significantly higher than that of C group (P < 0.05), the remodeling of bronchiole in A group was definite. (2) ROS (U/mg pro), GSSG (micromol/g pro) and total GSH in lung tissue of A group were significantly higher than that of C group (P < 0.01). The GSH/GSSG in lung tissue of A group was much lower than that of C group (P < 0.01), GSH in lung tissue showed no difference between A group and C group. (3) Immunohistochemistry indicated that Nrf2 protein and gamma-GCS protein were more positively expressed in A group than that in C group (P < 0.01). In situ hybridization discovered that the expression of gamma-GCS-h mRNA in lung tissue of A group was more positive than that of C group. (4) RT-PCR showed that the expression of Nrf2 mRNA was no difference between A group and C group (P > 0.05). (5) The activity of gamma-GCS of A group was (28 +/- 8)U which was significantly higher than that of C group (9 +/- 2)U (P < 0.01). (6) Linear correlation analysis indicated that in lung tissue of guinea pig with asthma there existed strongly positive relationship among ROS, GSSG and the expression of Nrf2, gamma-GCS mRNA, gamma-GCS protein, the activity of gamma-GCS, there existed strongly negative relationship among GSH/GSSG and the expression of Nrf2, gamma-GCS mRNA, gamma-GCS protein, the activity of gamma-GCS.

CONCLUSIONThere existed oxidative stress in lung of guinea pigs with bronchial asthma, which possibly positively regulated gamma-GCS via up regulating transcription factor Nrf2.

Animals ; Asthma ; metabolism ; Glutamate-Cysteine Ligase ; metabolism ; Guinea Pigs ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; RNA, Messenger ; genetics

OBJECTIVETo study the expression and significance of cyclin E, cyclin D1, CDK4 and p27 protein in esophageal squamous cell cancer (ESCC) and their correlation with tumor differentiation and lymph node metastasis.

METHODSExpressions of cyclin E, cyclin D1, CDK4 and p27 protein in 65 patients with ESCC were quantitatively detected by flow cytometry.

RESULTSThe expressions of cyclin E, cyclin D1, CDK4 in poorly-differentiated ESCC were higher than those in well-differentiated ESCC (P = 0.0275, 0.0001, 0.0174). The expression of p27 in poorly-differentiated ESCC was lower than that in well-differentiated ESCC (P = 0.0042). There was positive correlation between cyclin E and cyclin D1, cyclin D1 and CDK4, but negative correlation between cyclin D1 and p27. The expressions of all four proteins were not correlated with lymph node metastasis.

CONCLUSIONThe expressions of cyclin E, cyclin D1, CDK4 and p27 are closely related to tumor differentiation of ESCC. An imbalance between positive and negative control of cell cycling might be critical in the carcinogenesis of esophageal squamous cell cancer.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; secondary ; Cell Differentiation ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cyclins ; metabolism ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Flow Cytometry ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged

Enterotoxins ; analysis ; Humans ; Microbial Sensitivity Tests ; Staphylococcal Food Poisoning ; microbiology ; Staphylococcal Protein A ; analysis ; Staphylococcus aureus ; isolation & purification

Objective To analyze the prognostic factors of patients with adenocarcinoma of gastroesophageal junction (AGEJ) after radical resection. Methods In this retrospective study, 269 patients with AGEJ who underwent curative resection from March 2004 to June 2007 were enrolled.The survival curve was drawn by Kaplan-Meier method and the survival analysis was done by Log-rank test. Univariate and multivariate analyses of the prognostic factors of AGEJ were made by Cox model. Results The overall 5-year survival rate of 269 patients was 25.0 % with the median survival time of 22 months.The Siewert type and the number of positive lymph nodes could influence the survival rate (both P < 0.05). Among the 269 patients, 216 (80.3 %) were Siewert Ⅱ type, their median survival time was 30 and 12 months in the thoracic surgery group and the laparotomy group when the tumor diameter was 3-7 cm, the difference was statistically significant (χ2= 5.036, P= 0.025). Univariate analysis showed that tumor diameter, pT, pN, operation time, age and sex were significantly associated with survival rate (all P < 0.05). Cox multivariate analysis showed that patients with a more advanced tumor size suffered from a poorer prognosis (P< 0.05). The risk of postoperative death in patients with lymph node metastasis was 1.854 times that of patients without lymph node metastasis. Conclusions Patients with AGEJ are predominantly Siewert Ⅱ type, the transthoracic approach or thoracoabdominal approach are reasonable selections for these patients. Tumor diameter and pN are independent prognostic factors of AGEJ patients.

Objective To establish a real time PCR assay with a novel fluorescence quencher for identification of mutation of clarithromycin -resistance gene of Helicobacter pylori.Methods Two mutations of 23S rDNA gene in Helicobacter pylori,No.2142 and 2143,were chosen as targets for detection,and then the primers and the probe with a novel fluorescence quencher were designed.The genome DNA of Helicobacter pylori was extracted,and then detected by real time PCR reported here.Meanwhile,the specificity,reproducibility and sensitivity of the assay were evaluated.Finally,the real time PCR described here,the real time PCR based on TaqMan,and a sequencing assay were applied to detect 55 Helicobacter pylori strains isolated from clinical specimens,respectively.The results from three assays were compared with each other in order to further evaluate the applicability of this assay in clinic.Results It indicated that the mutation points related to clarithromycin -resistance,A2142G and A2143G,were identified by real time PCR with a novel fluorescence quencher rapidly and accurately.Moreover the coefficient of variation was less than 5%.The limit of detection was 100 copies/reaction.While this assay was applied directly to detect 55 Helicobacter pylori strains,the results were in accordance with those obtained from a TaqMan real time PCR and a sequencing assay,respectively.Conclusion The real time PCR described here was a simple,reliable and accurate approach and substituted for the TaqMan real time PCR for identification of two mutation points of clarithromycin -resistance,A2142G and A2143G in Helicobacter pylori.Thus,a novel tool for diagnosis of gene mutation was provided and the results might be regarded as a substantial evidence for clinical individual therapy.

The study established a UPLC-MS/MS method that is used for simultaneous determination nine major bioactive compounds of Dachengqi Tang in rat plasma. Using Aglient C18 column (2.1 mm x 50 mm,1.7 microm) was chromatographed, using methanol-5 mmol x L(-1) ammonium formate mobile phase gradient, elution 0.3 mL x min(-1). In the plasma pre-treatment process, not only the method of methanol and acetonitrile protein precipitation was investigated, and different factors extraction solvent, the type of the scroll time, the number and the type of extraction solvent, the extraction volume of the extraction solution of liquid-liquid extraction is investigated. Finally, with ibuprofen as an internal standard, using ethyl acetate liquid-liquid extraction method pretreatment blood, N2 dry reconstituted supernatant after centrifugation UPLC-MS/MS analysis, in electrospray ionization (ESI) negative mode, using multiple reaction monitoring mode for testing. The linear range of emodin, rhein, aloe-emodin, chrysophanol, magnolol, honokiol, hesperidin and hesperitin is 0.33-660, 0.40-792, 0.41-827, 0.34-680, 0.45-907, 0.46-927, 0.43-867, 0.34-683, 0.39-787 microg x L(-1) respectively, good linear relationship; and extraction recovery were greater than 69.39%, days after the day of the RSD is less than 15%. This method can be used to study the rat gastric large bearing gas after Dachengqi Tang, the simultaneous determination of nine components in plasma for its pharmacokinetics and efficacy material base to provide a theoretical basis.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods