The glycosylation heterogeneity of recombinant human pro-urokinase (pro-UK) was assessed using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Firstly, the source of heterogeneity was determined by measuring the M_r of intact protein before and after N-deglycosylation. Glycosylation sites and the proportion of O-glycopeptides then were determined at the peptide level. Finally, the N-glycans were confirmed and quantified using the N-glycan profile. Results show that the structural heterogeneity of pro-UK is mainly caused by glycosylation. All T₁₈ were fucosylated, and 6.4% of S_138/139 was O-glycosylated with two kinds of oligosaccharides with a ratio of 6.0% and 0.4% respectively. All N₃₀₂ positions were N-glycosylated by more than ten types of glycans, among which A2F and A3F accounted for 80% of the total. The assessment of glycosylation heterogeneity of pro-UK will provide a reference for quality standardization.

Carcinoma, Hepatocellular ; complications ; Hemangioma ; complications ; diagnosis ; immunology ; pathology ; Hepatitis B, Chronic ; complications ; Hepatocytes ; cytology ; pathology ; Humans ; Liver Cirrhosis ; complications ; Liver Neoplasms ; complications ; Male ; Middle Aged ; Platelet Endothelial Cell Adhesion Molecule-1 ; immunology ; Spleen ; immunology ; pathology ; Tomography, X-Ray Computed

Objective To investigate the combination of color Doppler flow imaging (CDFI) and transcranial Doppler (TCD) in the assessment of the hemodynamic changes of vertebral artery occlusion disease and their clinical value. Methods A total of 101 patients with vertebral artery occlusion detected by the combination of CDFI and TCD and confirmed by DSA were enrolled from January 2005 to January 2009. Taking the result of digital subtraction angiography (DSA) as a golden standard, The differences between the different types of the side of vertebral artery occlusion and contralateral vertebral artery on the extra- and intracranial segments were compared and analyzed in peak systolic velocity (PSV), end diastolic velocity (EDV), resistive indices (RI), pulsatility index (PI), spectrum morphology, and hemodynamics. Results No blood flow signals were detected by CDFI and TCD in patients of complete occlusion of the vertebral artery; the weak blood flow signals were detected by CDFI in patients of the occlusion in the intracranial segment of the vertebral artery. 3he blood flow signals after the establishment of collateral circulation in patients of the segmental occlusion were detected in the extraeranial segment or intraeranial segment of the vertebral artery. The PSV on the occluded sides of the extraeranial segments were decreased more significantly than that on the unoccluded sides (27.39 ± 12.44 cm/s vs. 62.61 ± 13.22 cm/s, P = 0.000); RI was significantly higher than the unoccluded sides (0. 99 ± 0. 21 vs. 0. 62 ± 0.07, P = 0. 000). When a vertebral artery had the segmental occlusion and the collateral circulation was established, the PSV, EDV, and PI of the intracranial segment of the vertebral artery on the occluded sides were decreased more significantly than those on the unoccluded sides, PSV were 37. 81 ± 12.28 cm/s and 73.17±30. 99 cm/s, respectively (P =0. 000), EDV were 17.58 ± 7.10 cm/s and 29.31 ± 12.94 cm/s, respectively (P = 0. 000), PI were 0.84 ± 0.22 and 1.01 ±0. 18, respectively (P=0. 000). The compard DSA showed that the sites of vertebral artery occlusion were different, There was significant difference in the Doppler flow velocity spectrum between the CDFI and TCD. Conclusions PSV, EDV, RI, and PI are the hemodynamic parameters of accurately assessing vertebral artery occlusion, and the combination of CDFI and TCD has significant value for the hemodynamie changes of different types of vertebral artery occlusion and the clinical comprehensive assessment.

Over the past three decades, more and more antisense drugs have been approved for marketing or clinical trails. Antisense technology has become the focus of pharmaceutical research due to its unique advantages in treating diseases and strong clinical development potential. There is a big difference from traditional small molecule chemical drugs, and macromolecular protein biological drugs. Antisense drugs are a very independent drug form. Antisense drugs were initially used to treat diseases with single gene mutations, but recently they have gradually begun to be used for the treatment of common diseases. Rational antisense drug design is crucial for disease treatment based on genetics. This paper reviews the latest progress in the field of action mechanism, chemical modification and delivery strategy of antisense drugs, and analyzes the current intractable problems. It is believed that with the resolution of these problems, the research of antisense drugs can reach a new level.

Objective To explore the correlation between Helicobacter pylori (Hp) infection and recurrent abdominal pain (RAP) in children over 6 years old and its relative treatment.Me-thods One hundred and eighty children over 6 years old with the diagnosis of RAP from Mar.2007 to Feb.2009 were selected,30 healthy children without RAP were taken as the healthy control group at the same period.14C-urea breath test (14C-UBT) was used to detect whether the patient was infected by Hp.The radical cure of Hp was given to the Hp-positive children with RAP,and the remission rate of children with RAP and the Hp negative rate were observed.Results The positive rate in RAP group was 58.33%,which was higher significantly than that in healthy control group(20.00%)(P0.05).The positive rate of those with and without bad eating habits were 45.56% and 12.78%,and the positive rate of with and without family gastrosis history was 49.44% and 8.89%,which both had significant difference between them(Pa

AIM: To explore the fundus fluorescein angiographic characteristics and relevant clinical significance of age-related macular degeneration(AMD).METHODS: Fundus fluorescein angiography (FFA) was performed on 149 eyes of 112 patients using Nikon NF-505 fundus camera.RESULTS: Out of 149 eyes, 90 eyes were atrophic AMD (60.4%), 59 eyes were exudative AMD (39.6%) which were further divided, according to the composition and location of lesion, into subfoveal choroidal neovasculari-zation (CNV)(7 eyes of classic type, 26 eyes of occult type, 9 eyes with disciform cicatrices, juxtafoveal CNV(2 eyes of classic type, 12 eyes of occult type), and extrafoveal CNV(3 eyes of occult type).CONCLUSION: FFA can show CNV of AMD patients and its quality and location, which is helpful to guide the treatment and evaluate the prognosis.

Fermentation for Eicosapentaenoic Acid(EPA) production by Nitzschia laevis at various temperature between 10℃ and 30℃ was investigated and the dynamics characteristics during fermentation process were also analyzed.Based on the results,a varying temperature nursing method of two stage control strategy is proposed:During the first stage,which comprises the delay phase and the initial index phase,the temperature is maintained at 25℃;then the temperature is shifted to 20℃ and kept up till the end of the fermentation process.By this method,a EPA content of 6.0% and a yield of 291.60 mg/L have been gained.These are 24.07% and 18.81% higher than that of fixed temperature(25℃) fermentation,respectively.

OBJECTIVE To investigate the risk factors,clinical characteristics and resistance of Staphylococcus aureus nosocomial infections so as to guide the treatment of S.aureus infection.METHODS To collect clinical materials of S.aureus nosocomial infection and analyze risk factors and clinical characteristics and detect sensitivity of isolated strains to antibacterial agents.RESULTS Severe underlying diseases existed among 73 cases of S.aureus nosocomial infections,82.19 percent of patients had received invasive interventions.Lower respiratory tract was the most common infective site.Seventy-nine strains of S.aureus were isolated,including 66 meticillin-resistant S.aureus(MRSA) and 13 meticillin-sensitive S.aureus(MSSA).S.aureus showed general resistance to many kinds of antibiotic drugs.The resistant rates of MRSA were much higher than those of MSSA(P

Objective To investigate the effect of tea polyphenols (TP) on renal damage in rats model induced by D-galactose. Methods Rats were injected with D-galactose (150 mg/kg?d),ip for 8 w,to induce renal damage. From the 3rd week,TP (150,75,37.5 mg/kg?d),aminoguanidine (150 mg/kg) and vitamin E (150 mg/kg) were administered with D-galactose for 6 w. After treatment,fasting blood glucose and 2 h blood glucose in oral glucose tolerance test were measured. The levels of HbA1C and fructosamine in serum,the activity of aldose reductase and content of advanced glycation end products (AGEs) in plasma and in kidney tissues and the activity of SOD,GSH-Px,and the contents of MDA in kidney tissues were measured,and 24h urinary protein,blood urea nitrogen (BUN) and serum creatinine (Cr) were detected. The apoptosis of renal cells were detected by flow cytometer. Results After treatment of D-galactose for 8 w,2h glucose level in oral glucose talerance test was increased significantly,the activity of aldose reductase and the content of AGES were increased significantly in blood. The levels of AGEs and MDA in renal tissues were also enhanced significantly. However,the activities of SOD and GSH-Px decreased. Additionally,the contents of 24h urine protein,BUN,Cr and the apoptotic rate of renal cells were increased significantly. High and middle dose of TP could can decrease the activity of aldose reductase in red blood cells,and inhibit the formation of glycation products in model rats induced by D-galactose. Also,TP could enhance the antioxidative activities and decrease the contents of AGEs and MDA in renal tissues. Mesnwhile,24h urine protein,BUN and Cr and the apoptotic rate of renal cells were increased significantly. Conclusion TP can inhibit glycation reaction induced by D-galactose and then protect renal from damage caused by glycation.

OBJECTIVETo study the genotoxicity of components of diesel engine exhausts with ethanol-diesel blending fuel. To provide scientific arguments to find more economical and less polluted fuels.

METHODSAmes test, comet assay and GC-MS technique were used to test the genotoxicity and 16 kinds of PAHs on diesel engine exhausts with different proportions of ethanol (E0, E5, E10, E20).

RESULTSBoth Ames test and comet assay were positive. It shows that diesel engine exhausts can lead to mutation and DNA damage, especially in pure diesel oil. But the content of 16 kinds of PAHs and DNA damage level decreased in exhausts of E5. With the increase of ethanol proportion in diesel oil, the content of 16 kinds of PAHs and DNA damage level increased.

CONCLUSIONCompared with pure diesel oil and high proportion of ethanol fuel, E5 can reduce the genotoxicity and the brake specific exhausts of PAHs.

Air Pollutants ; toxicity ; Air Pollution ; Carbon Monoxide ; Comet Assay ; DNA Damage ; Ethanol ; toxicity ; Gasoline ; toxicity ; Mutagenicity Tests ; Particulate Matter ; Vehicle Emissions ; toxicity