OBJECTIVE: To characterize the metabolism of genistin and study its enzymatic kinetics in rat liver microsome by HPLC-MS. METHODS: Genistin was incubated with rat liver microsomal incubation system. HPLC-MS method was used to characterize the metabolites. A metabolite generation method was established for quantitative analysis of genistein with sulfamethoxazole as internal standard. The enzyme kinetics parameters Vmax and Km was calculated by the GraphPad Prism 5.0 software. RESULTS: The metabolites in vitro incubation system was identified as genistein. The optimal time in rat liver microsomes incubation time of 40 min, the optimal protein concentration of 1 mg · mL^-1, a substrate concentration of 50 μmol · L^-1. The enzyme kinetics parameters of genistein were as follows; Vmax=(0.1042 ± 0.0033) μmol · min^-1 · mg(pro)^-1, Km=(28.96 ± 2.80) μmol · L^-1. CONCLUSION: The results indicate that genistin can be metabolited as the form of hydroxylation in rat liver microsome. Metabolite generation method is a reliable and simple method for determination of kinetic parameters of hepatic microsomal enzymes, and enzyme kinetic parameters obtained of genistin provide important parameters for further study.

Esophageal and Gastric Varices ; diagnosis ; etiology ; pathology ; Hemorrhage ; Humans ; Liver Cirrhosis ; complications ; pathology

Objective To explore the clinical features and operative treatment of floating shoulder injuries. Methods The clinical data of 36 patients with floating shoulder injuries that had been admitted to our hospital from June 1999 to June 2005 were retrospectively analyzed. The scapular neck fractures associated with clavicle fractures were in 31 cases and acromioclavicular joint dislocation in 5 cases. All cases were accompanied by associated injuries, in which rib fractures combined hemopneumothorax and/or lung contusion was 88.9%. The mean time from primary injury to the fracture operation was 9.6 days (range, 3 to 43 days). The clavicle fractures or acromioclavicular joint dislocation were dealt with firstly, then scapular neck fractures treated through modified Judet posterior approach. Thirty-three cases received internal fixation with both injuries of scapular neck and clavicle or acromioclavicular joint and 3 cases received internal fixation only in clavicle shaft. Results All fractures had been restored anatomical reduction in the target site. The mean followed up time was 19.7 months (range, 6 to 69 months). According to Constant and Murley's evaluation, the median score of functional results was 93% (mean 81.3%, range from 9% to 100%). Based on Herscovici's evaluation, 25 (69.4%) cases showed excellent results, 6 (16.7) good, 4 (11.1%) fair, and 1 (2.8%) poor. The recurrence of primary hemopneumothorax was found in 1 case, shoulder joint abduction weakness and subacromial space pain in 3 cases, delayed lesion of suprascapular nerve in 1 case, and posttraumatic shoulder joint instability secondary to arthritis in 1 case postoperation. Conclusion The double injuries with scapular neck and their suspensory device of clavicular shaft or acromioclavicular joint in floating shoulder injuries can hardly be corrected and reduced in three dimensional displacement of distal fracture unit without operation. It is an effective way for such unstable injuries to get good results after open reduction and internal fixation in early stage.

?Acute retinal necrosis syndrome ( ARN) is a serious eye disease, which caused by Herpes virus mostly, with unknown pathogenesis. Because of the aggressive progression, treatment of ARN is difficult, and the blindness rate is extremely high. Current treatment strategies are the combination of the drug therapy and the operative treatment. Drugs commonly used are antiviral drugs, glucocorticoids, and antiplatelet drugs, and the operative treatment includes laser photocoagulation and vitrectomy.

The rodipicolinate synthase gene was cloned from Nicotiana tabacum L.cv,which encodes the key enzyme in the synthesis of lysine,and then a vitro molecular reform reliefing feedback suppression was made. Some resistant seedlings were acquired using the mutated gene as selectable marker and analog of lysine as selectable reagent,and the detection results were positive by means of PCR and Real-Time PCR,however the phenotype is somewhat abnormal.

Adolescent ; Adult ; Automobile Driving ; China ; Electrocardiography ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Noise, Occupational ; adverse effects ; Occupational Exposure ; adverse effects ; Stress, Psychological ; etiology ; physiopathology ; Vibration ; adverse effects

BACKGROUNDWheat-dependent exercise-induced anaphylaxis (WDEIA) is a complex disease resulting from interaction of environmental and genetic factors. The aim of this study was to investigate the association of three single nucleotide polymorphisms (SNPs) (IL-4-C590T, IL-4RA A1727G and IL-10-A627C) with WDEIA.

METHODSSNP genotyping was conducted among the case subset composing 51 patients with WDEIA and four control subsets by sequencing DNA yielded from polymerase chain reaction (PCR). Statistical analysis of genotype/allele's frequencies between cases and controls were carried out through Fisher's exact test with the software of SPSS16.0.

RESULTSFor IL-4-C590T, there were statistically significant differences of genotype frequencies in case-control 1 (P = 0.03) and case-control 4 (P = 0.001) and statistically significant differences of allele frequencies in three case-control models (case-control 1: OR = 4.27 (95%CI = 1.40 - 13.07), P = 0.009; case-control 3: OR = 1.99 (95%CI = 1.13 - 3.50), P = 0.02; case-control 4: OR = 2.39 (95%CI = 1.49 - 3.84), P = 0.001). All other association studies showed no statistically significant (P > 0.05).

CONCLUSIONSIL-4-C590T may be related to the susceptibility of WDEIA, and the minor allele C might be a potential risk factor accounting for WDEIA. IL-4RA A1727G and IL-10-A627C might not be involved in the occurrence of WDEIA.

Adolescent ; Adult ; Aged ; Anaphylaxis ; genetics ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Exercise ; physiology ; Female ; Gene Frequency ; genetics ; Genotype ; Gliadin ; immunology ; Glutens ; immunology ; Humans ; Immunoglobulin E ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Triticum ; immunology ; Young Adult

OBJECTIVETo study the dynamic changes in the protein marker expression in the spermatogonial stem cells (SSCs) of mice at different ages by iTRAQ protein mass spectrometry and to screen new markers using the bioinformatic proteome database.

METHODSBased on the postnatal weeks, we divided 80 healthy male C57BL/6 mice into eight age groups of equal number, harvested their testicular tissues, extracted proteins following purification of the SSCs by compound enzyme digestion and magnetic-activated cell sorting. Then we analyzed and identified proteins using two-dimensional electrophoresis, protein mass spectrometry, and protein database information.

RESULTSTotally, 248,510 mass spectra were obtained from the MS experiment and 1132 proteins were identified. By the criteria of >1.2-fold for protein abundance difference and P value <0.05, we identified 298 differentially expressed proteins and 9 currently known makers of SSCs (PCNA, GFRalpha1, CDH1, Annexin A7, UCHL1, VASA, CD49f, CD29, and PLZf). Compara- tive analysis showed different expressions of the proteins in the SSCs of the mice of different ages, and the differences in the expressions of GFRalpha1, CD49f, and CD29 were consistent with the findings in other published literature. Ten proteins (P63, CD71, CD98, K19, ACE, K18, K15, K17, SH2, and SH3) were selected as SSC markers to be further studied.

CONCLUSIONThe proteins in SSCs are differentially expressed in mice of different ages. The technology of iTRAQ protein mass spectrometry can be used to analyze and compare the proteome information of mouse SSCs, obtain differentially expressed proteins in mice of different ages, and thus offers a new ap- proach to further analysis and study of the function and roles of these differential proteins.

Adult Stem Cells ; cytology ; metabolism ; Age Factors ; Animals ; Biomarkers ; analysis ; metabolism ; Cell Separation ; methods ; Electrophoresis, Gel, Two-Dimensional ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred C57BL ; Proteins ; analysis ; metabolism ; Spermatogonia ; cytology

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Evidence-Based Medicine ; Humans ; Lung Neoplasms ; complications ; therapy ; Neoplasms ; complications ; drug therapy ; Pain, Intractable ; drug therapy ; Phytotherapy ; Stomach Neoplasms ; complications ; therapy

Objective To compare the dosimetric verification results of Varian Portal Dosimetry and Matrixx,and to assess the reliability of the clinical application of electronic portal imaging device (EPID) verification.Methods Varian TrueBeam linear accelerator,which was equipped with a 120-leaf multileaf collimator and an amorphous silicon EPID,as well as portal dose prediction software.IBA I′mRT Matrixx ion chamber array was used.EPID algorithm configuration,dose calibration,and testing before use were performed.The sliding-window protocol was used.There were 77 patients with tumors involving the head and neck (mainly nasopharyngeal carcinoma),mediastinum,abdomen,and pelvic cavity were selected.The verification plan of the portal dose was created with a source-detector distance of 100 cm,and the gantry angle was kept the same as the treatment plan.The verification plan was carried out in the TrueBeam machine,and the data were collected at the same time by EPID.Comparison between the measured and calculated dose images was performed,and the evaluation standard was gamma index (3%/3 mm).The paired t-test was used for difference analysis.Results For the 77 patients,the Gamma passing rates of both methods were above 97%.Except for head and neck carcinoma were a significant difference between the results of dosimetric results using EPID and Matrixx in intensity-modulated radiotherapy (P=0.018) other remaining all P> 0.05.Conclusions The dosimetric verification results of EPID are consistent with those of Matrixx.EPID can be used for dosimetric verification,and Matrixx ion chamber array can be used only in case of a low Gamma passing rate.