Accidents, Occupational ; Humans ; Hydrogen Sulfide ; poisoning

Objective To establish a stable and rapid separation and purification method for Astragalus membranaceus (Am) pathogenesis-related protein-10 (AmPR-10) using an automatic intelligent protein purification system AKTA Avant 25, and analyze its physiochemical and biological activity. Methods Am was extracted by Tris-HCl buffer. The crude extract was captured by anion exchange chromatography, and finely separated by hydrophobic chromatography and gel filtration chromatography. The relative molecular weight of AmPR-10 was measured by MALDI-TOF/TOF mass spectrometry, the protein identification was determined by mass spectrometry and MS/MS Ion Search, the glycoprotein identification was estimated by periodic acid-Schiff method, and the ribonuclease activity and effect factors were analyzed by agarose gel electrophoresis. Results The electrophoretically pure AmPR-10 was obtained by three-step purification of Q Sepharose Fast Flow, Butyl Sepharose High Performance and SuperdexTM 75 10/300 GL from the crude extraction. The relative molecular weight of AmPR-10 was 16 801. AmPR-10 was highly homologous to PR-10 and has no carbohydrate chains. Incubated at 56 ℃ for 30 min, AmPR-10 exhibited significant ribonuclease activity to total RNA of mammalian cells. The activity was insensitive to NaCl, pH value and mental ions, and weekly inhibited by 0.5 mol/L NaCl, pH 9.0, Mg2+ and Co2+. The activity was the same at EDTA as high as 20 mmol/L. Conclusion The three-step method of exchange chromatography-hydrophobic chromatography-gel filtration chromatography, a stable and rapid separation and purification method of AmPR-10, can be applied for other Chinese herbs. AmPR-10 might play an important role in resistance against virus.

Adolescent ; Adult ; Aged ; Cadherins ; genetics ; metabolism ; physiology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Ki-67 Antigen ; genetics ; metabolism ; Male ; Microtubule-Associated Proteins ; genetics ; metabolism ; physiology ; Middle Aged ; Neoplasm Invasiveness ; physiopathology ; Pituitary Neoplasms ; pathology ; physiopathology ; Young Adult

Aged ; Antigens, CD34 ; analysis ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Solitary Fibrous Tumors ; metabolism ; pathology

Specific primers 1CaA/1CaB for full cry1C gene in Bacillus thuringiensis subsp.colmeri strain 15A3 were designed. The 4.0kb PCR product included the whole ORF and regulation region of cry1C gene. This PCR product was linked with shuttle vector pHT315 by two cloning steps. The recombined plasmid pHT-1C was electroporated into Bacillus cereus 9509, a kind of bacteria that beneficial to crops. The transformant could produce bipyramidal-shaped parasporal inclusions. The 60kD protein band was detected by SDS-PAGE. The bioassay result showed that the cry1C gene transformant of Bc 9509 had insecticidal activity to Spodoptera exigua.

Objective To detect the levels of AKAP12 methylation by methylation‐specific high‐resolution melting curve(MS‐HRM ) in peripheral blood in patients with colorectal cancer and investigate its clinical significance .Methods We used MS‐HRM technology to detect the levels of AKAP12 methylation in peripheral blood in 60 lung cancer patients ,and analyzed the relationship between the levels of AKAP12 methylation and pathological parameters of lung cancer patients .Results 34(56 .7% ) of the 60 lung cancer patients were found to be methylated at the AKAP12 promoter region by MS‐HRM ,the methylation levels of 18 cases ranged between 1% -20% ,14 cases ranged between 20% -60% ,2 cases ranged between 60% -100% .There was no significant differences between the levels of AKAP12 methylation and lung cancer patients′age and gender(P>0 .05) .However ,it was signifi‐cantly higher in the patients with high pathological stage and differentiation degree (P<0 .05) .Conclusion AKAP12 promoter re‐gion methylation was related to tumor progression and malignant degree .

Objective Despite regular treatment,antineutrophil eytoplasmie antibodies(ANCA)asso- ciated systemic vasculitis(AASV),in which the role of Fc?Rs has been established,are still associated with significant long-term mortality and remain an important cause of end-stage renal failure.ANCA plays an im- portant role in the pathogenisis of primary systemic small vessel vasculitis(PSV)by their potential to activate neutrophils.Because the interaction between ANCA and its receptors on the Fc portion of immunoglobulins (Fc?R)on neutrophils is essential in the activation process,we investigate the inhibitory,effect of tg19320 on ANCA induced activation of neutrophils,which is a tetrameric tripeptide that interferes with IgG/Fe?Rs in- teraction.Methods We prepared tg19320 by solid-phase peptide syntbesis.The binding between tg19320 and human IgG was assessed by enzyme-linked immunosorbent assay.The biological activity of tg19320 to intefere with FcF?receptor recognition was identified by rosette formation assay.ANCA IgG was prepared from the sera of active Wegener's granulomatosis(WG)and microscopic polyangiitis(MPA)patients.Neu- trophils isolated from the blood of healthy volunteers were primed with TNF-?(2 ng/ml)and then incubated with ANCA IgG(200?g/ml),or pretreated with tg19320(2.5 mg/ml)and then added with ANCA IgG.Su- peroxide burst of neutrophils was determined by Ferri-cytochrome reduction assay.Results We found that tg19320 bound tightly to human IgG in a dose dependent manner and the inhibition of the rosette formation between SRBC-IgG and U937 cells was statistically significant(20.3% vs 53.2%,P

1.0,achieved the standard request of Disinfection Technology standard.RESULTS Through EOW disinfection,the bacteria were not found on the object surfac of transports and conformed to the object surface disinfection request of Disinfection Technology Standard.CONCLUSIONS Applying EOW to disinfection of polluted instrument recycling vehicle for clinical purpose can fulfill the State standards completely;EOW has strongly germicidal activity,also with characteristics as rapidity,safety and environment security,without any irritant odor.These results indicate a widely potential use of EOW as in hospital disinfection supply center,especially,with a positive significance in health care for pediatric patients.

OBJECTIVETo explore the effect and action mechanism of moxibustion on healing of cutaneous wound in rats.

METHODSTwenty-four SD rats were selected and made into linear full-thickness skin injury model. With randomized digital table, rats were randomly divided into a treatment group and a model group, 12 cases in each one. Then according to treatment time, each group was again divided into a 1d group, a 3d group and a 7d group, 4 cases in each one. The moxibustion at injured skin was applied in the treatment group, 30 min per time, once a day. Hematoxylineosin (HE) staining method was adapted to measure growth status of capillary and number of vascular endothelial cell; immunohistochemical method was used to measure the expression of vascular endothelial growth factor (VEGF).

RESULTSThe wound healing indices in the treatment 7d group were higher than those in the model 7d group on both the 4th day and 8th day after treatment (both P < 0.05). The number of capillary in the treatment 1d group and 3d group was higher than that in the model 1 d and 3 d groups (both 1 < 0.05). The number of capillary in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The number of vascular endothelial cell in the treatment 3d group was higher than that in the model 3d group (P < 0.05). The number of vascular endothelial cell in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of number of vascular endothelial cell between the treatment 1d group and model 1d group was not significant (P > 0.05). Positive cells accumulated score of V EGF expression in the treatment 3d group was higher than that in the model 3d group (P < 0.05). Positive cells accumulated score of VEGF expression in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of positive cells accumulated score of VEGF expression between the treatment 1d group and model 1d group was not significant (P > 0.05).

CONCLUSIONMoxibustion could improve the healing of skin wound in rats, which could be related with regulating vascular endothelial cell and VEGF in wound tissue at different time.

Animals ; Endothelial Cells ; metabolism ; Female ; Humans ; Male ; Moxibustion ; Rats ; Rats, Sprague-Dawley ; Skin ; injuries ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Wounds and Injuries ; genetics ; metabolism ; therapy

Objective To investigate the effects of simvastatin on lipopolysaccharides (LPS)induced upregulation of Lp-PLA2 in human peripheral blood monocytes-macrophages and the related mechanisms. Methods Peripheral blood monocytes of healthy volunteer were isolated and incubated for 2-3 days. Monocytes were incubated with various concentrations of LPS for 6 h or with 1 μg/ml of LPS for different times in LPS group. In simvastatin group and MAPK inhibitors groups, cells were pre-treated with simvastatin ( 10-2 - 10-7mmol/L)or various MAPK inhibitors ( 10 μmol/L SB203580, 20 μmol/L U0126,and 20 μmol/L SP600125) before LPS co-incubation. Lp-PLA2 activity was measured by chromometry, LpPLA2 mRNA expression was detected by RT-PCR. Protein expressions of Lp-PLA2 and p38MAPK and phosphorylated p38MAPK were examined by Western blot. Results ( 1 ) LPS significantly upregulated LpPLA2 mRNA and protein expression, as well as the enzyme activity in a time and concentration dependentmanner, which could be significantly attenuated by simvastatin in a time and concentration dependent manner. (2) Simvastatin significantly reduced LPS-induced p38MAPK phosphorylation. The p38 MAPK inhibitor SB203580, but not MEK1/2 inhibitor U0126 and JNK inhibitor SP600125, completely prevented LPS-mediated up-regulation of Lp-PLA2 at protein level. Conclusion This study demonstrated that LPS significantly upregulated Lp-PLA2 mRNA and protein expression, as well as the enzyme activity in a time and concentration dependent manner via Rho-p38MAPK pathway, which could be significantly suppressed by simvastatin.