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

Objective To investigate the diagnosis value of laparosco-pie examination in ascites.Methods The data of twenty-three cases with ascites of unknown origin undergoing laparescopie examination and pathologic examination were retrospectively analyzed from May 2004 to August 2008.Results Twenty-two eases were definitely diagnosed by laparoscopie exploration,the positive rate was 95.6%.One ease re-mained unknown.Among them,18 eases were tuberculous peritonitis; 3 eases were metastatic adenocarcino-ma; one ease was peritoneal mesothelioma.No obvious postoperative complications occurred.Conclusion Laparoscopie examination is a safe method with hige definite diagnostic rate,which has important values in diagnosis of ascites of unknown origin.

Objective To investigate the feasibility and safety of laparoscopic cholecystectomy(LC) in patients with chronic renal dysfunction at azotaemic stage.Methods Clinical data of 7 patients accompanying chronic renal dysfunction at azotaemic stage treated with LC between May 2004 and September 2006 were analyzed retrospectively.The operation was performed under general anesthesia and endotracheal intubation.The CO2 pressure was maintained at 9～12 mm Hg.The LC was conducted with 3-port technique in 5 patients and 4-port technique in 2.Results The LC was completed smoothly in all the 7 patients.Patients' renal dysfunction was not aggravated.There was no significant difference between pre-and post-operative time in levels of blood urea nitrogen(11.92?4.06 mmol/L vs 12.16?3.76 mmol/L;t=0.50,P=0.633) and blood creatinine(208.62?134.37 ?mol/L vs 204.20?125.53 ?mol/L;t= 0.51,P=0.626).As compared with preoperative levels,the creatinine clearance rates were not significantly changed at 2 weeks after operation in 3 patients(40.03 ml/min vs 45.61 ml/min;32.28 ml/min vs 38.93 ml/min;56.72 ml/min vs 51.60 ml/min).Follow-up checkups for 4～20 months(mean,10 months) showed no aggravation of renal dysfunction.Conclusions Laparoscopic cholecystectomy is a feasible and safe procedure for patients with chronic renal dysfunction at azotaemic stage.

Objective The practical evaluation of Rosco Disk Diffusion method on the clinical antifungal susceptibility test for yeast-like fungi. Methods The Fluconazole susceptibility test was detected by Rosco Disk Diffusion method and NCCLS M27-A broth macrodilution method with 76 yeast-like fungus strains isolated from clinical specimens. Three standard strains were used as the quality control. Results Coincidence rate of the two methods was 90.8%,The sensitive strains detected by one method didn't show resistance in another detection method, and the resistant strains detected by one method yet didn't show sensivity in another.Conclusion Rosco disk diffusion method can be used in clinical detection instead of NCCLS M27-A broth macrodilution method.

Objective To study the feasibility of laparoscopic cholecystectomy accompanied by simultaneous umbilical hernia repair.Methods Twenty-four adult patients received laparoscopic cholecystectomy accompanied by simultaneous umbilical hernia repair in our hospital from November 2003 to April 2008.An incision was made in navel defect and entered the abdominal cavity under a direct vision.LC was performed by 3-trocar technique.Umbilical hernia was repaired by Mayo' style.Results The operation was successfully completed in all the cases.The operation time ranged from 35 to 65 minutes (mean,48 minutes),the blood loss was 5-30 ml(mean,16 ml),and hospital stay after the operation was 3 -4 days.No patient dereloped wound infection or skin necrosis.No recurreme was found during a 1-to 54-month follow-up(mean,28.6 months).Conclusions Laparoscopie cholecystectomy accompanied by simultaneous umbilical hernia repair is a safe and effective procedure.Mayo repair is simple and effective for umbilical hernia with less fascial defects.

ObjectiveTo synthesize metal complex that exhibits superoxide dismutase (SOD)-like activity and determine its structure.MethodsThis chemical compound was synthesized by means of reflux in laboratory.Its structure was examined by infrared spectra (IR),nuclear magnetic resonance hydrogen spectra(H-NMR) and mass spectra (MS).The activity of the SOD was determine by xanthine oxidase (XOD).ResultsThe IR indicted that this complex showed a typical absorption band of -OH,C=N and benzene. The H-NMR reveals the existence of metal ion manganese (Mn3+ ,paramagnetic). The molecular weight was 365, which was obtained by EI-MS analysis. And this complex showed SOD-like activity.ConclusionA metal complex with low molecular weight and SOD-like activity has been synthesized.

Objective To study the protective effects of the compound with low molecular weight of anti free radical on brain ischemic and hypoxic injuries in rats.Methods The model of brain ischemic and hypoxic injuries in rats was established by unilateral carotid artery ligature just for 2 h. The compound was injected 30 minutes after the rats act on hypoxic state (10% O2+90%N2) for 1 h. Then the serum of rats was separated and the value of the superoxide dismutase (SOD) activity and malondiade (MDA) were determined, and the pathological changes were observed in brain tissue.Results Compared with rats in the model group, the compound decreased the concentrations of MDA in serum (P<0.05), and raised the level of SOD (P<0.05) significantly in therapy group. Otherwise, the pyramidal cells remained organized order, with only a few cells degenerated occasionally in rats of therapy group. Conclusion The compound has SOD like activity. It can scavenge the free radical and inhibit peroxidation of lipid, so it can prevent brain cells from the damage.

Objective To investigate the spherical diopter and astigmatism change of humans at sitting and supine position.De- sign Prospective case series.Participants 96 eyes of 52 patients (spherical diopter from-2.50 D to-10.00 D,astigmatism diopter from -0.75 D to-4.50 D) were selected.Methods The subjects were examined with NIKON portable retinomax at sitting and supine posi- tion,respectively.Main Outcome Measures The spherical diopter,cylinder diopter and axis change were analyzed statistically.Re- sults Spherical diopter at supine position (-5.31?3.43 D) was a little higher than that at sitting position (-5.27?3.24 D) statistically(P= 0.25),and cylinder diopter at sitting position (-2.27?1.24 D) and at supine position (-2.35?1.19 D) was no statistically difference (P= 0.20).The axis of astigmatism changed from-16?to +18?.Axis change was within 2?in 52.1% eyes,6?-10?in 5.2%,over 10?in 3.1%. The change of axis rotation tended to counter-clockwise in the right eye and clockwise in the left eye.Conclusions Eye rotation at sit- ting and supine position may cause the astigmatism axis change.It may be one of the main factors affecting the results of LASIK.

Objective To evaluate the expression of Sp3 gene of peripheral blood mononuclear cells (PBMC) in multiple sclerosis (MS) patients in Guizhou and the relationship between Sp3 gene expression and immunological function. Methods Two pairs of primers were used to amplify cDNAs generated from 31 MS patients and 30 healthy controls. The serum levels of sIL-2R were measured in 27 patients with MS and 30 healthy controls by sandwiched ELISA. Results The deficient expression of Sp3 gene in MS patients was significantly higher than that in control (41.9% ( 12/31 ) vs 6. 7% (2/30) ,x2 =7. 133 ,P =0. 008). The sIL-2R levels in MS patients were significantly higher than those in control (( 2788.5 ± 1079. 8 ), ( 1270. 7 ± 489. 4) μg/L, t = 6. 170, P = 0. 001 ). The concentration of sIL-2R in MS with negative ((3364.0 ± 1252.3) μg/L) and positive((2450.0 ± 827.0) μg/L) expression of Sp3 gene were significantly increased compared with control (F = 32. 059, P < 0. 05 ). The sIL-2R levels were significantly rising in MS patients with negative expression of Sp3 gene compared with MS patients with positive expression of Sp3 gene ( q = 4. 213, P < 0. 05 ). Conclusions A remarkable deficient expression of Sp3 gene in PBMC has been found in MS patients in Guizhou. sIL-2R may take part in the process of MS. The expression of Sp3 gene is not affected by immune state, however, MS patients with Sp3 deficient expression tend to have a more serious impairment in immunological functions.

Objective To discussed the relationship between multiple sclerosis(MS)and the genepolymorphism of HLA hoping that these results would be useful for further pathogeny studies,diagnoses,therapy and prognosis estimation of MS.Methods HLA-Ⅱ alleles in 32 patients with MS,36 nonimmunological neurological disease controls and 30 healthy controls,were identified by polymerase chain reaction-specific sequence primers(PCR-SSP)methods.Results The gene frequency of HLA-DR16 (7/32,1/36,0/30),DR11(7/32,3/36,1/30)and DQB1*0502(10/32,6/36,4/30)in patients with MS were higher than those in the 2 control groups.The gene frequency of DQB1*0601(8/32,12/36,17/30)in the patients with MS was lower than the controls.However,only the HLA-DR16 had significant difference (χ2=7.398,RR=17.94,P=0.011;χ2=5.52,RR=9.8,P=0.022).Conclusion HLA-DR16 alleles may be associated with the susceptibility to MS in Guizhou Province.