With development of bio-technique, more and more proteins were applied as clinical approaches. However, the protein homogeneity, especially the N-glycosylation limited the further research and application of these protein drugs. The analysis method for N-glycans is believed to be critical in protein drugs development. To enhance the N-glycans isolation efficiency and accelerate the pretreatment, a new strategy was built on ultrafiltration-devices. New methods increased the isolation efficiency of N-glycans containing N-acetylglucosa mine with 10%-20%. The degrading of N-glycans containing sialic acids was also minimized with this method. 20%-100% more N-glycans with sialic acids were isolated. The pretreatment was finished within 30 min. Coupled with HPLC-HRMS, an effective and reliable strategy designed for protein drugs N-glycans analysis were developed.
Glycoproteins ; chemistry ; Glycosylation ; Polysaccharides ; chemistry ; Ultrafiltration ; instrumentation

Ascites ; therapy ; Humans ; Treatment Outcome ; Ultrafiltration ; instrumentation

OBJECTIVES: To investigate the interference of postmortem hemolysis on the detection of creatinine and whether ultrafiltration can reduce the interference. METHODS: A total of 33 non-hemolyzed whole blood samples from the left heart were collected. Hemolyzed samples with 4 hemoglobin mass concentration gradients H1-H4 were artificially prepared. Ultrafiltration was performed on each hemolyzed sample. Creatinine concentrations in non-hemolyzed serum (baseline serum), hemolyzed samples and ultrafiltrate were detected. Bias (B), Pearson correlation and receiver operator characteristic (ROC) of baseline creatinine concentration between before and after ultrafiltration were analyzed. RESULTS: As the hemoglobin mass concentration increased, B of the hemolyzed samples in the H1-H4 groups gradually increased, the |B| was 2.41(0.82, 8.25)-51.31(41.79, 188.25), reaching a maximum of 589.06%, and there was no statistically significant between the creatinine concentration and the baseline creatinine concentration (P=0.472 7, r=0.129 5). After ultrafiltration of hemolyzed samples, the interference of creatinine concentration in ultrafiltrate was significantly reduced, the |B| was 5.32(2.26, 9.22)-21.74(20.06, 25.58), reaching a maximum of 32.14%, and there was a positive correlation with baseline creatinine concentration (P<0.05, r=0.918 2). In the hemolyzed samples of H3 and H4 groups, there were 7 false-positive samples and 1 false-negative sample; in the ultrafiltrate samples, there were no false-positive sample and 1 false-negative sample. ROC analysis results showed the hemolyzed samples were lack of diagnostic value (P=0.117 5). CONCLUSIONS The postmortem hemolysis significantly interferes creatinine detection results of blood samples, ultrafiltration can reduce hemolysis-induced interference in postmortem creatinine detection.
Humans ; Creatinine ; Hemolysis ; Ultrafiltration ; Serum ; Hemoglobins

OBJECTIVE: We determined the influence of 3-OMD in the protein binding of levodopa to estimate the effect of 3-OMD on the penetration of levodopa into brain. METHOD: P-glycoprotein in the brain may serve to limit drug penetration into the brain. Because it is not available as an experimental substance, but has similar binding properties with alpha 1 acid glycoprotein(AGP), we used AGP in this study. Additionally, we used blood plasma to see the affinity of plasma binding of levodopa. The final concentration of chemicals used in this study were 125, 250, 500, 1000, 2000, 4000 microgram/l for levodopa and 0, 1250, 5000, 10,000 microgram/l for 3-OMD, 1 mg/l for AGP. The free fraction of levodopa in blood plasma and AGP were separated by ultrafiltration method and determined by beta-counter, respectively. RESULTS: We found that levodopa did not bind with AGP, but only 22-24% from 125 microgram/l to 4000 microgram/l of it bound with blood plasma. The addition of 3-OMD to the blood plasma did not significantly change the binding of levodopa. CONCLUSIONS: We can conclude that 3-OMD does not influence the penetration of levodopa into brain. These small amount of the binding does not expect to influence to other drugs on the binding with plasma.
Brain ; Drug Interactions ; Levodopa* ; P-Glycoprotein ; Plasma* ; Protein Binding ; Ultrafiltration

In order to evaluate the peritoneal transport characteristics in Korean non-diabetic and diabetic end- stage renal disease patients, peritoneal equilibration test(PET) proposed by Twardowski et al. were performed on patients who had been on continuous ambulatory peritoneal dialysis(CAPD) for 2 to 6 months. The results were as follows : 1) Fifty four patients(including 24 diabetics) on CAPD were studied with a mean age of 48.7 years. And male/female ratio was 1 : 1.08. 2) In non-diabetics, the dialysate to dialysate prior to infusion ratio for glucose(D/D0 glu) at 2-, and 4-hour dwell times were 0.61+/-0.09, and 0.39+/-0.10, and the dialysate-to-plasma ratio for creatinine (D/P cr) at 2-, and 4-hour dwell times were 0.40+/-0.11, 0.63+/-0.12, respectively. 3) In diabetic patients, D/D0 glu at 2-, and 4- hour dwell times were 0.60+/-0.09, 0.39+/-0.08, respectively, and D/P cr at same dwell times were 0.50+/-0.08, and 0.71+/-0.08, which were significantly higher than in non-diabetics(p<0.05). 4) According to the two-hour plasma glucose concentration, diabetic patients were subdivided into hyperglycemic(>or=150mg/dL) and normoglycemic(<150mg/ dL) patients. The values of D/Pcr at 2-, and 4-hour dwell times in hyper-glycemic patients were signficantly higher than in non-diabetic patients (D2/P2 cr : 0.50+/-0.09 vs. 0.40+/-0.11, D4/P4 cr : 0.72+/-0.07 vs 0.63+/-0.12, respectively, p<0.05). 5) Net ultrafiltration did not differ between any of subgroups. 6) In non-diabetic patients, the ranges of D4/P4 cr and D4/D0 glu for high, high average, low average, and low transporters were defined as D4/P4 cr : 0.87-0.75, 0.75-0.63, 0.63-0.51, 0.51-0.39, D4/D0 glu : 0.19-0.29 0.29-0.39, 0.39-0.49, 0.49-0.59, respectively, which were remarkably simliar as suggested by Twardowski et al. In conclusion, the creatinine and glucose transfers assessed by dialysate-plasma ratio of creatinine and glucose are remarkably similar between Korean and North American patients. And the creatinine transport rate in Korean diabetic patient is higher than non-diabetic patient while ultrafiltration is achievable in non-diabetic patient.
Blood Glucose ; Creatinine ; Glucose ; Humans ; Peritoneal Dialysis, Continuous Ambulatory* ; Ultrafiltration

OBJECTIVETo investigate the effect of removing bacterial endotoxin in the key processes of Reduning injection.

METHODThe content of bacterial endotoxins was detected by kenitic-turbidimetry and the removal efficacy was studied before and after using 0.8% of activated carbon and ultrafiltration with molecular weight cut-off of 10 x 10(3).

RESULTThe adsorption rate of bacterial endotoxins was 78.7% by using activated carbon, while the removal efficacy of bacterial endotoxins was 99.6% with ultrafiltration membrane at cut-off molecular weight 10 x 10(3).

CONCLUSIONThe key technology can effectively guarantee the safety of Reduning injection.

Sclerosing peritonitis is an uncommon complication of continuous ambulatory peritoneal dialysis (CAPD) and can lead to small bowel dysfunction involving abdominal pain, progressive loss of ultrafiltration, and small intestinal obstruction. Peritoneal thickening, in which calcification can develop, often starts as a small plaque which gradually becomes larger. We report a case of CAPD-related calcifying peritonitis.
Abdominal Pain ; Intestinal Obstruction ; Peritoneal Dialysis, Continuous Ambulatory ; Peritonitis* ; Ultrafiltration

OBJECTIVEThe UF process of model systerm of Huanglian Jiedu decoction is researched in the study to lay a foundation for the exploring of the substance foudation of membrane fouling, and for the the optimal design of membrane technology in the production of Chinese drugs preparation.

METHODUsing the membrane flux, fouling degree of membrane, preserving degree and adsorption capacity of effective materials as guide line, trying to find out the influence of macromolecule materials on small molecules in Huanglian Jiedu decoction.

RESULTThe two small molecules berberine and gardenoside in Huanglian Jiedu decoction have high permeation rate. But for the reasons of interaction of macromolecule, the permeation rate of small molecules reduced greatly, membrane fouling and membrane flux declined intensify.

CONCLUSIONStarch and pectin are the two main macromolecule materials in Huanglian Jiedu decoction which cause membrane fouling and flux falling.

Objective To investigate the treatment effect of hollow fiber ultrafiltration technology on hemolytic samples and the differences between IgE concentration and serum concentration before hemolysis in ultrafiltrate. Methods The 33 postmortem blood samples of non-frozen corpses within 72 hours after death were collected, 4 mL blood was taken from each case, among which 1 mL was centrifuged to get serum, and the remaining 3 mL blood was frozen-thawed 3-5 times to cause complete hemolysis. The 2 mL hemolytic samples were processed by hollow fiber ultrafiltration to obtain ultrafiltrate. The hemoglobin concentration in serum, complete hemolytic sample and ultrafiltrate was determined by Van-Zij solution-cyanated methemoglobin assay method, and the total IgE in serum and ultrafiltrate was determined by electrochemical luminescence method. Results The hemoglobin concentration in ultrafiltrate was significantly lower than that in complete hemolytic samples （P<0.05）. There was a good correlation between the total IgE detection values of ultrafiltrate and serum （r=0.984）. The difference between the serum and the value of IgE in ultrafiltrate after correction had no statistical significance, and the differences between the two in positive rates had no statistical significance （P>0.05）. Conclusion Ultrafiltration technology has a good treatment effect on complete hemolytic samples, and the correction value of ultrafiltrate detection is close to the serum level before hemolysis, and therefore, it can be applied to the detection of total IgE of frozen corpse hemolytic samples.
Autopsy ; Hemolysis ; Humans ; Immunoglobulin E/analysis* ; Serum ; Ultrafiltration

Peritoneal ultrafiltration failure is a common reason for peritoneal dialysis (PD) withdrawal as well as mortality in PD patients. Based on the three-pore system, inter-cellular small pores and trans-cellular ultra-small pores (aquaporin-1) are mainly responsible for water transfer across the peritoneum. Both small and ultra-small pores-dependent water (free water) transport decline accompanied with time on PD, with more significant decrease in free water, resulting in peritoneal ultrafiltration failure. The reduction of free water transport is associated with fast peritoneal solute transfer, reduced crystalloid osmotic gradient due to increased interstitial glucose absorption, and declined osmotic conductance to glucose resulted from impaired aquaporin-1 function and peritoneal interstitial fibrosis. The decline of small pore-based water is mainly because of fast loss of crystalloid osmotic gradient, decrease of hydrostatic pressure mediated by peritoneal vasculopathy, as well as reduced absolute number of small pores. The current review discusses the advance on pathogenesis of acquired peritoneal ultrafiltration failure in long-term PD.
Humans ; Peritoneum ; Ultrafiltration ; Dialysis Solutions ; Peritoneal Dialysis/methods* ; Water ; Glucose