The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.
Amino Acid Sequence ; Chromatography, High Pressure Liquid ; methods ; Interferons ; standards ; Mass Spectrometry ; methods ; Molecular Weight ; Oxidation-Reduction ; Peptide Mapping ; Protein Processing, Post-Translational ; Reference Standards

AIM: To explore the frequency of drug injection of alloxan diabetes on the established model of rabbit.METHODS: Thirty-six healthy rabbits, weighing 2-2.5kg, were randomly divided into one time drug injection group (group A, n=12), two times drug injection group (group B, n=12) and three times drug injection group (group C, n=12). Each rabbit was injected with a total amount of 150mg/kg of alloxan. Fasting blood glucose was measured. The success rate and death rate of each group were also calculated.RESULTS: The success rate of diabetic rabbit model in group B was higher than that in group A (P<0.01) and its death rate was lower than that of group A (P<0.01); the success rate of diabetic rabbit model in group C was highest and the death rate was the lowest in three groups(P<0.01). CONCLUSION: Multiple administration of alloxan can improve success rate in establishing diabetic rabbit model with decreased death rate and increased stability.

AIM: To evaluate the effects of 23G vs 20G pars plana vitrectomy ( PPV ) combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation for macular epiretinal membrane with cataract. ·METHODS: Totally 45 eyes of 45 patients with macular epiretinal membrane and cataract were enrolled in this retrospective non-randomized controlled clinical study. All eyes were treated with PPV combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation. There were 20 eyes in 23G PPV group, and 25 eyes in 20G PPV group. The best corrected visual acuity ( BCVA ) , intraocular pressure (IOP), counting of corneal endothelial cells ( CEC) and central retinal thickness ( CRT ) were examined before surgery. BCVA results were converted to the logarithm of the minimum angle of resolution ( LogMAR ) visual acuity. All operations were performed by the same doctor. Operation time for vitrectomy and membrane peeling, average ultrasound energy ( AVE) and effective phacoemulsification time ( EPT ) were recorded. BCVA and CRT were observed postoperatively at 30d and 90d, counting of CEC was observed postoperatively at 90d. IOP was observed postoperatively at 1d and 7d. ·RESULTS:The mean operation time for vitrectomy were 12. 57± 1. 35min in 23G group and 17. 30 ± 1. 19min in 20G group. The difference was statistically significant ( t =-12. 488, P<0. 01). There were no statistical significances in operation time for membrane peeling, AVE and EPT between 23G and 20G groups ( t=-0. 68,-1. 186,-0. 737, P=0. 500, 0. 242,0. 465). On 1d after surgery, IOP in 23G group was lower than that in 20G group, the difference was statistically significant (t= -2. 345, P=0. 024). The BCVA and CRT of the two groups both improved after operations. There were no statistically significant differences between two groups in terms of IOP, BCVA, and CRT ( F = 0. 465, 1. 895, 0. 689; P = 0. 499, 0. 176, 0. 411). IOP, BCVA and CRT were significant statistical different in different time-point within each group ( F=291. 245, 103. 06, 665. 402, P<0. 01 ). Different surgical methods of 23G and 20G had interactive effects on IOP with different time points ( F = 13. 245, P<0. 01 ), but different surgeries had no interactive effects on BCVA and CRT with different time points (F=1. 212, 2. 293;P=0. 283, 0. 129). The counting CEC in 23G group was more than that in 20G group postoperatively at 90d, the difference was statistically significant (t=2. 049, P=0. 048). ·CONCLUSION: The 23G PPV combined with internal limiting membrane peeling, phacoemulsification, intraocular lens implantation for macular epiretinal membrane with cataract is effective. Compared with 20G PPV, 23G PPV has advantages in operation time for vitrectomy and counting CEC. But lower IOP is likely in 23G PPV on 1d after surgery

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.

Objective To observe the effect of transplanting the adipose-dervived stem cells(ADSCs) on free radical metabolism and immune function of rat aging model induced by D-galactose from fasiaology perspective;to explore a new method for anti-aging. Methods The ADSCs were cultured in vitro. Thirty male SD rats were randomly divided into 3 groups: normal control group(A), aging model group(B) and treat group(C). Ten rats in each group. Rats in B and C groups were injected D-galactose continually into make the sub-acute aging model rats.After 8-week injections of D-galactose;R3ats in group C were injected ADSCs(3×10~6/ml) through caudal vein. After 2-week transplantions of ADSCs, T-SOD, CuZn-SOD, MDA, NO, IL-2 and spleen index levels in serums of each group were detected and compared among the three groups. Results Compared with the A group, the SOD, NO, IL-2 level and spleen index in serum in group B decreased significantly, while the contents of MDA increased significantly. Compared with group B, the SOD, NO, IL-2 level and spleen index in serum in group C had been improved, and the contents of MDA decreased significantly. Conclusion Transplanting ADSCs can improve the antioxidant ability and strengthen the cellular immune function of aging rats.Further more, it can delay the ageing procedure induced by D-galactose in rats.

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.

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

OBJECTIVETo compare clinical outcomes between two suturing methods using non absorbable materials through drilling the bone and suturing anchors for the treatment of complete rupture of the deltoid ligament.

METHODSFrom January 2009 to January 2013, 58 hospitalized patients with ankle fracture combined with complete rupture of the deltoid ligament were treated with suturing using non absorbable materials through drilling the bone or suturing anchors. There were 29 patients who received suturing treatments using non absorbable materials through drilling the bone (Group A), including 18 males and 11 females, with an average age of (39.76 +/- 11.81) years old. According to the Lauge-Hansen classification, 12 patients had supination external rotation (SER) injuries with IV degree, 5 patients had pronation external rotation (PER) injuries with III degree, 10 patients had PER injuries with IV degrss, and 2 patients had pronation abduction injuries with III degree. There were 29 patients who received treatments with suturing using anchors (Group B), including 14 males and 15 females, with an average age of (41.79 +/- 13.28) years old. According to the Lauge-Hansen classification,9 patients had SER injuries with IV degree, 6 patients had PER injuries with III degree,13 patients had PER injuries with IV degree, and 1 patient had pronation abduction injuries with III degree. All the patients were treated with open reduction and internal fixation, as well as reconstruction of deltoid ligaments to restore the stability of the medial ankle structures. The clinical examination, imaging evaluation, American society for ankle surgery (AOFAS) ankle-hindfoot score and visual analogue scale (VAS) were used to evaluate the clinical results after operation, and the results of the two groups were compared and analyzed statistically.

RESULTSThe follow-up duration of the 58 patients ranged from 23 to 40 months,with an average of 27.3 months. All the patients had fracture union, and the mean healing time was 12.3 weeks (ranged, 10 to 17 weeks). There were no incision complications and ankle instability. There were no significant differences between two groups in AOFAS (P=0.666) and the VAS (P=0.905).

CONCLUSIONTreatments of complete rupture of the deltiod ligaments with the two suturing methods get similar good clinical effects, but the suturing using non absorbable materials through drilling the bone has several advantages such as reducing the financial burden of patients, saving social medical resources and avoiding the shortcoming in difficult removal of anchor suture.

Adolescent ; Adult ; Ankle Fractures ; surgery ; Ankle Injuries ; surgery ; Ankle Joint ; surgery ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; Humans ; Lateral Ligament, Ankle ; injuries ; surgery ; Male ; Middle Aged ; Young Adult

Structure of a recombinant chimeric anti-CD20 IgG1 monoclonal antibody was verified by the application of high-performance liquid chromatography-mass spectrometry (HPLC-MS)and N-terminal sequencer. Molecular masses, N-terminal sequences and peptide maps of the antibody treated with different reagents and enzymes were measured. Results indicate that the amino acid sequences of light and heavy chains and 10 disulfide bonds were consistent with theoretical structure. By comparison of molecular masses and peptide maps for the fully glycosylated and deglycosylated samples, the N-linked glycosylation site was identified. The method is simple, rapid, precise, and could be referred to the quality control and structure determination of other IgG1 products.
Amino Acid Sequence ; Antibodies, Monoclonal ; chemistry ; Antigens, CD20 ; immunology ; Chromatography, High Pressure Liquid ; Glycosylation ; Immunoglobulin G ; chemistry ; immunology ; Immunoglobulin Heavy Chains ; chemistry ; Immunoglobulin Light Chains ; chemistry ; Mass Spectrometry ; Molecular Weight ; Peptide Mapping ; Recombinant Proteins ; chemistry ; Trypsin ; chemistry

AIMTo explore the function state of protein tyrosine phosphatase (PTP) SHP-1 and CD45 under high partial pressure of oxygen (Po2) in lymphocyte.

METHODSPrimarily cultured rat splenic lymphocytes were treated with different pressure-duration of oxygen that led to promotion or inhibition of cell function. The catalytic activities, protein expression and tyrosine phosphorylation of SHP-1 and CD45 were determined.

RESULTSThe activity of SHP-1 was decreased after high Po2 treatment no matter what pressure-duration was, while the activity of CD45 was decreased only after high PoF that led to inhibition of lymphocyte function. The proteins expressed and tyrosine phosphorylation levels of two enzymes had no alteration in most treatment groups.

CONCLUSIONThe decrease of PTP catalytic activities might be caused by their structures modification by high Po2 induced reactive oxygen species. SHP-1 and CD45 might be one of the key points of action through which high PO2 exerts its effects on lymphocytes.

Animals ; Cells, Cultured ; Leukocyte Common Antigens ; metabolism ; Lymphocytes ; enzymology ; metabolism ; Male ; Oxygen ; metabolism ; Partial Pressure ; Phosphorylation ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism