To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.
Animals ; Benzaldehydes ; administration & dosage ; blood ; pharmacokinetics ; Benzofurans ; administration & dosage ; blood ; pharmacokinetics ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Caffeic Acids ; administration & dosage ; blood ; pharmacokinetics ; Catechols ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, Liquid ; methods ; Diterpenes, Abietane ; administration & dosage ; blood ; pharmacokinetics ; Hydroxybenzoates ; administration & dosage ; blood ; pharmacokinetics ; Injections, Intravenous ; Lactates ; administration & dosage ; blood ; pharmacokinetics ; Phenanthrenes ; administration & dosage ; blood ; pharmacokinetics ; Plant Preparations ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Tandem Mass Spectrometry ; methods

OBJECTIVETo further articulate the conceptual framework of sub-health by way of literature review.

METHODSPublished papers and relevant literature about sub-health were collected, the representative publishing about theoretical exploration and practical surveys were focused for analyzing and assessing the category of sub-health, investigative objects, assessing tools, conceptual framework, diagnostic criteria and its operability, foundation support, etc.

RESULTSSub-health conditions were categorized into three kinds in most documents, namely, the somatic, the mental and the social adaptative sub-health, and the relevant symptoms were classified into three levels. However, contents regarding the homogeneous issues, such as meaning and definition of sub-health, its diagnostic criteria and operability, in different documents were discrepant.

CONCLUSIONDiscrepancy exists between the theoretical and practical studies of sub-health. And detailed description is needed to articulate the problems respectively on specific and non-specific sub-health. The perspicuity of conceptual framework is the basis of sub-health researches. The authors recommended that to form the framework the four-level hierarchic structure should be adopted; in developing the assessing tool for sub-health diagnosis, besides the basic assessment on concrete conditions, the differential evaluation on the belonging of overall manifestations, subjective complain or objective symptom, should also be paid attention to.

The yeast Sacchromyces cerevisiae is most widely used for producing bioethanol in alcoholic industry due to its higher ethanol yield and fermentation rate. However, the toxic effect of accumulated ethanol is one of the main factors, which limit high ethanol production. Thus, investigating the mechanisms of yeast ethanol tolerance will provide the basis for solving the industrial problem. This article reviewed the mechanisms of Sacchromyces cerevisiae ethanol tolerance focusing on its cell physiological behaviors, structure and biochemical composition, as well as its genetic basis.

Objective To investigate the relationship between the endoscopic characteristics of chronic gastritis,duodenitis,peptic ulcer and their histopathologic findings in children,and explore the relationship between Helicobacter pylori(Hp) infection and the severity of histopathologic changes of gastroduodenal mucosa in children. Methods Three hundreds and sixty-six children with chronic upper gastrointestinal symptoms who were examined by gastroscopy were enrolled.The gastric and duodenal mucosal biopsy specimens of all the patients were studied histopathologically. ResultsTypes of chronic gastroduodenal diseases in all these patients were: chronic gastritis(n=206,56.3%),chronic gastritis combined with duodenitis(n=112,30.6%),chronic gastritis combined with peptic ulcer(n=48,13.1%).There was chronic inflammation in gastric mucosa and duodenal bulb mucosa in all the cases examined by histopathologic examination.Hp infection was found in 106 cases(28.96%).The gastric mucosal inflammation was more severe in those with Hp infection than those without(P0.05).The were significant differences in the incidences of inflammation activity,atrophia and nodulus lymphaticus of gastric mucosa between those with Hp infection and those without(P

Objective To study the treatment efficacy of auricular point sticking for primary dysmenorrhea in college students, and to seek a convenient effective treatment method for primary dysmenorrhea.Method A total of 144 female college students with primary dysmenorrhea were randomized into an auricular point sticking group, a medication group, and a blank control group to receive the corresponding intervention. The dysmenorrhea intensity score and traditional Chinese medicine symptoms score for dysmenorrhea were used for observation.Result After the intervention, the recovery rate and total effective rate were respectively 64.6% and 91.7% in the auricular point sticking group, versus 39.6% and 70.8% in the medication group. There was a significant difference in comparing the therapeutic efficacy between the auricular point sticking group and medication group (P<0.05).Conclusion Auricular point sticking can produce a significant efficacy in treating primary dysmenorrhea in female college students, without adverse effects and convenient, and has a content long-term efficacy.

OJECTIVE: To explore the changes of serum IgE and tryptase caused by anaphylactic shock rats and discuss the relation to PMI and preservative environment of corpse and specimen. METHODS: Rats were used for establishing anaphylactic shock models and randomly divided into room temperature group, refrigeration group, frozen group, manual hemolysis group, specimen preservation group. And the control group was also established. The blood samples were collected after rats were sacrificed. The degree of hemolysis was graded according to the color of the upper layer of the serum. The mass concentration of IgE and tryptase in each group was detected by ELISA. RESULTS: The levels of serum IgE and tryptase in anaphylactic shock dead rats were higher than that of the control group. Room temperature and frozen made obviously differences on the levels of serum IgE and tryptase with various PMI. The levels of serum IgE and tryptase in refrigeration group showed relatively stable. The levels of serum tryptase and IgE were elevated with differently increasing hemolysis. The levels of serum IgE and tryptase showed no obvious changes during the specimen kept under different temperature conditions for 25 days. CONCLUSION Serum IgE and tryptase obviously increased in anaphylactic shock rats. However, the levels were influenced by PMI and environmental temperature, especially under the conditions of room temperature and frozen.
Anaphylaxis/blood* ; Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulin E/blood* ; Rats ; Temperature ; Tryptases/blood*

Objective To investigate the value of tumor-stroma ratio(TSR) in evaluating the prognosis of patients with triple-negative breast cancer (TNBC).Methods A total of 87 patients with TNBC were selected from January 2010 to December 2014 in the Second People's Hospital of Yibin.The TSR was detected by Hematoxylin eosin staining,the patients with TSR ＜50％ were included in the stroma-rich group,and the patients with TSR≥50％ were included in the stroma-rare group.The influencing factors of tumor-free survival and overall survival in TNBC patients were analyzed by COX regression model.The effect of TSR on tumor-free survival and overall survival in TNBC patients was further analyzed by Kaplan-Meier method.Results Among the 87 TNBC patients,there were 38 patients with TSR≥50％ (stroma-rare group) and 49 patients with TSR ＜ 50％ (stroma-rich group).The 5-year tumor-free survival rate and overall survival rate of the 87 TNBC patients was 52.9％ (46/87) and 56.3％ (49/87) respectively.Single factor COX regression analysis showed that the tumor diameter staging,lymph node metastasis,the number of tumors and TSR were significantly correlated with the overall survival rate and tumor-free survival rate of patients with TNBC (P ＜ 0.05).Multivariate COX regression analysis showed that the tumor diameter staging,lymph node metastasis,the number of tumors and TSR were independent risk factors for overall survival rate and tumor-free survival rate of patients with TNBC (P ＜ 0.05).Kaplan-Meier survival analysis showed that the tumor-free survival rate and the overall survival rate of TNBC patients in the stroma-rare group were 68.4％ (26/38) and 73.7％ (28/38),and they were 40.8％ (20/49) and 42.9％ (21/49) in the stroma-rich group.The Log-Rank test showed that the overall survival rate and disease-free survival rate of the TNBC patients in the stroma-rare group were significantly higher than those in the stroma-rich group (x2 =7.788,9.799;P ＜ 0.05).Conclusion TSR is significantly associated with the prognosis of TNBC patients,and it is an independent risk factor for the prognosis of TNBC patients.TSR can be used as an indicator for evaluating the prognosis of TNBC patients.

WRKY transcription factor is one of the Zinc finger proteins which contains a highly conserved WRKY domain and is a family of the plant-specific transcription factor. The plasmid pET28a-SmWRKY1 harboring Salvia miltiorrhiza WRKY1 (SmWRKY1) gene was successfully transformed and expressed in Escherichia coli BL21 (DE3). The conditions on protein expression of SmWRKY1 in E. coli, including induction duration, temperature, IPTG concentration and the E. coli concentration were optimized. The results showed that the highest protein expression of SmWRKY1 was obtained at 24 hours after the E. coli was cultured in the presence of 0.2 mol x L(-1) IPTG at 20 degrees C with A600 values of 1.0-1.5. This recombinant histidine-tagged protein was expressed at 2.454 g x L(-1) as inclusion body, which was first extracted using urea, and then purified by Ni2+ affinity chromatography and identified by SDS-PAGE. The expression of SmWRKY1 in E. coli was further confirmed by western blotting analysis.
Blotting, Western ; Cloning, Molecular ; DNA-Binding Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; chemistry ; genetics ; metabolism ; Molecular Weight ; Plant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Salvia miltiorrhiza ; genetics

Hemangioma ; pathology ; Humans ; Joint Diseases ; pathology ; Male ; Middle Aged ; Synovial Membrane ; pathology

Fatal anaphylactic shock is common in forensic practice. However, it is difficult to diagnose for lacking specific pathological and morphologic changes in forensic autopsy. The application of some biochemical indicators is of great significance. This paper reviews the biological characteristics of some biochemical indicators and detection methods. The forensic application, problems and prospects of these indicators are also introduced in details. The stable biochemical indicators, IgE, tryptase and chymase, show great potential and advantages in the identification of fatal anaphylactic shock in forensic medicine.
Anaphylaxis/metabolism* ; Autopsy ; Biomarkers ; Chymases ; Forensic Medicine ; Humans ; Tryptases