Proteomics is a powerful subject focusing on large-scale study of protein structures and functions. A complete enzymatic digestion of protein complexes is the key step in modern high-resolution and high-throughput mass spectrometry (MS)-based identification and quantification. To achieve MS analysis, both peptide sample pretreatment and data acquisition are prerequisite in proteomic studies. In this paper, we summarized both the enzymatic proprieties of three common proteolytic enzymes, Trypsin, Lys-C and Glu-C, the optimization of multi-enzyme combination and an advanced sample pretreatment in proteomics research.
Enzymes ; chemistry ; Mass Spectrometry ; Proteins ; chemistry ; Proteomics ; methods ; Trypsin ; chemistry

Angiofibroma ; diagnosis ; Diagnostic Errors ; Humans ; Male ; Middle Aged ; Pharyngeal Neoplasms ; diagnosis ; Polyps ; diagnosis

In this study, three methods for identification of E.coli were compared. The conventional method was employed to select and identify the suspicious E.coli isolates from a fecal sample. PCR based ARDRA analysis was then carried out to distinguish these E.coli isolates, E.coli MG1655 and other bacterial species. All the potential E.coli isolates and E.coli MG1655 had the identical ARDRA banding pattern while the other bacterial species showed the different patterns.The result indicated that the ARDRA analysis was consistent with the traditional method for identification of E.coli and could be the practical method for distinguishing E.coli from other intestinal bacterial species. The ERIC-PCR analysis provided abundant polymorphism between different E.coli isolates, and might be a powerful approach for elucidating the genetic diversity among isolates of the same species.

Background Researches showed that the content of nitric oxide (NO) and nitric oxide synthetase (NOS) increases in blood,aqueous humor and tear of cataract patient.But the function of NO and NOS in cataract formation is still elusive.Objective The aim of this study was to explore the prevention and treatment effect of NOS inhibitor,1-nitro-arginine methyl ester (L-NAME),on galactose cataract.Methods Sixty clean three-week-old Wistar rats were equally and randomly divided into 3 groups.0.9％ Normal saline solution (30 ml/kg) was subcutaneously injected every day for 30 days in the rats of the control group,and 50％ of D-galactose solution (30 ml/kg) was used in the rats of the model and L-NAME group at the same way.L-NAME eye drops was simultaneously administered in the L-NAME group 3 times per day for 30 days.The eyes of the rats were examined under the slit lamp in 10,20 and 30 days,and the degree of lens opacification was scored.Lenses of the rats were obtained at the end of this experiment for the detect of NO,NOS contents.Flow cytometry was used to assay the caspase-3 level of rat lens.Repeated measurement two factor analysis of variance was used to analyze the difference of lens opacification scores in different groups and different time points,and one-way ANOVA was used to analyze the differences of NO,NOS and caspase-3 contents in lens among the groups.Results Lens opacification appeared in 10 days after injection of 50％ D-galactose solution in the rats of the model group and L-NAME group.Lens opacification score was higher among the different groups and different time points (Ftime =435.251,P =0.000 ;Fgroup =395.120,P=0.000).NO content in the lens was (0.45±0.15) μmol/g,(2.67 ± 0.47) μmol/g and (1.68±0.34) μmol/g in the control group,model group and L-NAME group,showing a significant difference (F=58.872,P=0.000).The NOS contents in the lens was (0.0160±0.0020) U/ml,(0.0370±0.0040) U/ml and (0.0270±0.0010) U/ml in the control group,model group and L-NAME group,showing a significant difference (F =66.174,P=0.000).Caspase-3 contents in the lens was (339.4 ± 37.9),(697.7 ± 46.5) and (650.7 ± 53.1),Showing a significant difference among them (F =100.005,P =0.000).Conclusions The increase of NO,NOS and caspase3 levels are associated with lens opacification.Topical administration of L-NAME eye drops can down-regulate NOS content in lens,reduce the NO formation and inhibit the apoptosis of lens epithelial cells.

CAS (Chrome azurol S) assay was a universal method for detecting the bacterial siderophores, and Sugar-Asn liquid medium has been applied to the studies on siderophores from Pieudomonas. In this paper, Asp has been substituted for Asn, and MSA-CAS agar plate was developed by integrating the MSA (sugar-Asp) medium and CAS bright blue dye, which has been used in the universal CAS assay. On the aspect of siderophores detection , 8 strains of 7 species from Pseudomonas had been screened on MSA-CAS agar plates and universal CAS assay respectively. The results showed that MSA-CAS agar was higher-sensitive and lower basic fluorescent than universal CAS assay.

A novel stable blue agar plate which is convenient to preparing and more effective than the universal chrome azurol sulfonate (CAS) assay established by Schwyn and Neilands was designed by replacing MM9 growth medium and pipes with certain concentrate of phosphorus buffer solution which pH could be stabled at 6.8. It is more suitable for screening over- siderophores production bacteria. Since OD_ 630 of the sample is usually out of the range of spectrophotometer with CAS assay solution when quantifying the siderophores and the outcome is not steady,the measuring wavelength had been changed to 680 nm corresponding to the middle of max absorbance and the correlation between siderophores concentrations and OD was unchanged. But the detecting sensitivity is elevated by enlarged the absorbance differences among samples with different productivity of siderophores at 680 nm .

OBJECTIVETo evaluate the therapeutic effect of Liujunzi decoction combined with Zuojin pills in treating the radioactive duodenitis and their mechanism, and compare with clinical routine acid suppressants combined with mucous membrane protective preparations to study the mechanism of their efficacy.

METHODAccording to the study of Williams J P and characteristics of duodenitis, and by reference to the radiation enteritis modeling standard, we took the lead in establishing the mouse radioactive duodenal injury model. The model mice were randomly divided into the control group (n = 26), traditional Chinese medicine (TCM) group (n = 16) and the western medicine (oral administration with famotidine 0.5 mL + almagate suspension 0.3 mL per mouse, once a day) group (n = 16). After the standard administrating, such objective indexes as general condition, weight, changes in health score, pathology and expression of inflammatory factors were observed to evaluate the efficacy.

RESULTThe radioactive duodenitis model of mice was successfully established with 12 Gy. Mice in the control group suffered from weight loss, anorexia, low fluid intake, loose stools, and occasionally mucous bloody stool, poor spirit, dim fur, lack of exercise and arch back. Mice in drug intervention groups were generally better than those in the pure irradiation group. The IL-6, IL-1beta, TNF-alpha mRNA expressions in spleen and mesenteric lymph node tissues in TCM and western medicine groups showed a declining trend compared with the control group. Their concentrations in peripheral blood serum also slightly changed. The TCM group revealed notable advantage in reducing inflammatory factors. The microscopic observation showed that a better mucosa repair in intervention groups than the pure irradiation group. The improved Chiu's scoring method showed a statistical significance in the difference between TCM and western medicine groups (P < 0.05).

CONCLUSIONLiujunzi decoction combined with Zuojin pills could treat acute radiation enteritis, regulate organic immunity, and inhibit acute injury, promote local tissue repair, with the potential to resist such adverse effects as radiation intestinal fibrosis. The regulation of inflammatory factor release is one of efficacy generation mechanisms.

Animals ; Cobalt Radioisotopes ; adverse effects ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Duodenitis ; blood ; drug therapy ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Mice ; Mice, Inbred BALB C ; Mucous Membrane ; drug effects ; radiation effects ; Radiation Injuries, Experimental ; blood ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

Adenocarcinoma, Bronchiolo-Alveolar ; metabolism ; pathology ; Antigens, CD34 ; metabolism ; Diagnosis, Differential ; Female ; Hemangioendothelioma, Epithelioid ; metabolism ; pathology ; surgery ; Hemangioma ; metabolism ; pathology ; Hemangiosarcoma ; metabolism ; pathology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Middle Aged ; Vimentin ; metabolism

Purple bacteria are extensively used in bacterial photosynthesis research. This review describes the structure of light-harvesting I, light-harvesting II and reaction center of photosynthetic apparatus of pur- ple bacteria and discusses the regulation mechanisms of photosynthesis (PS) gene transcription, with an emphasis laid on the transcriptional regulation of PS gene by PpsR/AppA system.