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

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.

Objective To compare the primary treatment efficacy of radiofrequency ablation and surgical resection in patients with solitary hepatocellular carcinoma (HCC) which the diameter is ≤5 cm.Methods Databases were searched for comparative studies on radiofrequency ablatiom vs surgical resection published from 2005 to 2012.A Meta-analysis was performed using a randomized or fixed effect model to compare the treatment efficacy between radiofrequency ablatiom and surgical resection.Results Five studies fulfilled the criteria and were included.For HCC patients whose single tumors' diameter is ≤5 cm,radiofrequency ablation was equivalent to surgical resection for 1-,3-and 5-year overall survivals (P ＞ 0.05).However,surgical resection was superior to radiofrequency ablation in 1-,3-and 5-year disease-free survivals and there' s significant difference (P ＜ 0.05).Higher local recurrence rate was associated with radiofrequency ablation than surgical resection.Conclusions For HCC patients whose single tumors' diameter is ≤5 cm,radiofrequency ablation can achieve comparable overall survival as surgical resection,though with higher recurrence rate and lower disease-free survival.

OBJECTIVE:To provide reference for clarifying the pharmacodynamic material basis of anti-inflammation effect of Uygur medicine Hyssopus cuspidatus. METHODS:HPLC was conducted to establish the fingerprint of 8 extracts with different po-larities. Using macrophage RAW264.7 as object,the inhibitory effect of extracts with different polarities on inflammatory factor ni-tric oxide(NO),TNF-α and IL-6 in supernatant of cell culture medium was detected,the differences of anti-inflammatory activity in vitro were compared. Grey correlation analysis method was used to analyze the spectrum-effect relationship of peak area of common peaks and anti-inflammation activity. RESULTS:The fingerprint of 8 extracts with different polarities showed obvious differences. Anti-inflammation in vitro results suggested that 30% ethanol extract had the strongest inhibitory effect on inflammatory cytokines in vitro. There were 14 common peaks in the established HPLC fingerprints,the 5 common peaks that was closely related to the in-hibitory effect of inflammatory factors in vitro were peak 8,6,5,10,13,respectively;peak 8 was rosmarinic acid. CONCLUSIONS:The established fingerprint and its spectrum-effect relationship with anti-inflammation activity in vitro can provide certain reference for its pharmacodynanic material basis study.

Objective To evaluate the viability of residual tumor after insufficient thermal ablation of rabbit VX2 liver tumor and investigated the efficacy of sorafenib as an adjuvant therapy.Methods Twenty-one rabbits were implanted with VX2 tumor to establish orthotopic liver tumor models.They were allocated randomly into 3 groups:control (n =7),ablation (n =7),and combination treatment (n =7).Microwave coagulation therapy was conducted with 20 W for 1 min and viable tumor tissue remained at the periphery.A laparotomy was performed in the control group.Sorafenib was given at 20 mg/kg/d during the following 10 days in the combination treatment group,and saline was given to the control and ablation group.Tumor volume was recorded before and after treatment,immunohistochemistry detected CD31 and proliferating cell nuclear antigen (PCNA) expression,and the micro-vessel density (MVD) and proliferation index (PI) were calculated accordingly.Results Ten days after insufficient ablation,tumor volume of the ablation group was larger than that of the control group (P ＜0.05).The MVD and PI of residual tumor were higher compared with those of the control group (P ＜ 0.05).With adjuvant therapy of sorafenib after insufficient ablation,tumor volume showed a decrease on the 10th day compared with tumors undergoing insurfficient ablation alone (P ＜ 0.05).The MVD and PI of residual tumor were lower than those of the ablation group (P ＜0.05).Conclusion Insufficient thermal ablation promotes residual tumor progression,but adjuvant therapy of sorafenib serves as an effective way to suppress the overgrowth and neovasculation of the residual tumor.

Objective To investigate the clinical effect of blocking nail in treatment of hypertrophic nonunion after interlocking intramedullary nailing of femoral shaft fractures.Methods A retrospective case series study was made on 11 patients with hypertrophic nonunion following interlocking intramedullary nailing of femoral shaft fractures from January 2006 to February 2016.There were 8 males and 3 females,aged (32.4 ± 6.8) years (range,19-48 years).Comminuted fractures were noted in three patients,long oblique or spiral fractures in two,multi-segment fractures in two,and transverse fractures in four.Time interval between this surgery and intramedullary nailing of femoral shaft fractures was (11.5 ±3.1) months (range,6-18 months).All blocking nails were inserted into pulp cavity of distal ends of the site of the bone nonunion.Fracture healing time,wound healing and postoperative complications were detected.Outcome was evaluated using the Visual Analogue Scale (VAS) and Oswestry Disability Index (ODI).Results All patients were followed up for (27.2 ± 4.6) months (range,24-36 months).Bone union was observed within (9.8 ± 3.4) months except that bone nonunion occurred in two cases requiring locking plate fixation and autologous bone grafting to provide union,with the healing rate of 91％.No wound infection,nail breakage and nail loosening occurred after operation.At postoperative 3 months,4 months,5 months,6 months,12 months and 24 months,VAS and ODI differed significant from the preoperative detections (P ＜ 0.05).Conclusion Blocking nail technique is associated with increased stability of intramedullary nails and fracture ends,low incidence of complications and high bone healing rate in treatment of hypertrophic nonunion after femoral interlocking intramedullary nailing.

OBJECTIVE: To study mutations in the GJB2 gene in Kazak patients with nonsyndromic hearing impairment from Xinjiang. METHOD: One hundred and ninety-three cases of Kazak from the Xinjiang region, including 97 cases of hearing loss and 96 cases of normal people, were performed mutational analysis of the GJB2 coding region by PCR-direct sequencing. RESULT: Eight kinds of mutation were found in the encoding region of hearing impairment group:12 cases of 35 delG homozygous, 5 cases of 79G>A homozygous, 8 cases of 79G>A heterozygous, 1 case of 79G>A heterozygous and 608T>C heterozygous, 5 cases of 79G>A heterozygous and 341A>G heterozygous, 4 cases of 235 delC heterozygous, 2 cases of 341A>G heterozygous, 1 case of 439T>G heterozygous, 1 cases of 457G> A heterozygous, 2 cases of 521G>A homozygous. Four kinds of mutations found in the normal group were confirmed as common polymorphic mutation. CONCLUSION The study suggests that the GJB2 gene mutation of the Kazak deaf population in Xinjiang has ethnic and regional characteristics. There is a rather high carrier frequency of GJB2 mutation of Kazak patients in Xinjiang. In this study the 35 delG mutation is a common mutation of Kazak patients.
Adolescent ; Adult ; Child ; Child, Preschool ; China ; Connexin 26 ; Connexins ; genetics ; Deafness ; genetics ; Ethnic Groups ; genetics ; Humans ; Infant ; Mutation ; Young Adult

OBJECTIVE:To conduct component analysis for the 40％ ethanol eluate of Hyssopus officinalis,and investigate its improvement effect on inflammation in asthmatic mice. METHODS:The 40％ ethanol eluate of polyamide resin column was col-lected,and HPLC-high resolution mass spectrometry was used for the component analysis of 40％ ethanol eluate of H. officinalis. Totally 72 mice were randomly divided into blank group (normal saline),model group (normal saline),dexamethasone group (positive control,1.6 mg/kg) and 40％ ethanol eluate of H. officinalis high-dose,medium-dose,low-dose groups (200,100,50 mg/kg),12 in each group. Except for normal group,mice in other groups were intraperitoneally injected 0.2 mL ovalbumin (OVA)for sensitization in 0,14 d and intragastrically administrated in 25-31 d,once a day. After administration,2 mg/mL OVA was dropped in nose for 7 d. After 24 h of last dropping in nose,tumor necrosis factor α(TNF-α),interleukin-4(IL-4),interfer-on-γ(IFN-γ)levels in bronchoalveolar lavage fluid(BALF)were detected;pathological changes in lung tissue were observed. RE-SULTS:Totally 11 compounds were identified,the relative percentage content of 40.89％. The main components were rosmarinic acid,luteolin 7-O-β-D-rhamnoserhamnose (1→6)-α-D-pyran glucoside,hyperoside,etc. Compared with blank group,TNF-α, IL-4 levels in BALF in model group were increased,IFN-γ level was declined,and IL-4/IFN-γ ratio was enlarged(P<0.01);lung tissue was seriously damaged,there was infiltration of inflammatory cells around the blood vessels. Compared with model group, TNF-α,IL-4 levels in BALF in dexamethasone group,40％ ethanol eluate of H. officinalis high-dose,medium-dose groups were declined,IFN-γ level was increased,and IL-4/IFN-γ ratio was reduced (P<0.01);pathological changes in lung tissue were improved. CONCLUSIONS:The established analysis method can effectively analysis the chemical components of 40％ etha-nol eluate of H. officinalis,which has certain regulatory effect on releasing inflammatory factors and reducing inflammatory lesions in lung tissue of mice with bronchial asthma.

Objective In this paper we analyzed the major risk factors of hemorrhagic fever with renal syndrome(HFRS) as well as its clinical manifestation,so as to provide a basis for clinical diagnosis and effective control of HFRS in Xi'an.Methods The method of retrospective study was used to collect clinical data of hospitalized patients with HFRS between 2005 and 2010 in the Hospitals for Infectious Diseases in Xi'an city,Zhouzhi and Huxian counties and then clinical manifestations of the patients with HFRS were analyzed and classified according to the national standards for clinical symptoms and test indicators of HFRS.HFRS patients matched by the ratio of 1 ∶ 2 healthy human controls were retrospectively investigated in order to obtain risk factors relevant to HFRS incidence using casecontrol study.Odds ratio(OR) method was used for single factor study.While for the multifactor study,we took the conditional Logistic regression approach.We also built models for both studies.Factor with OR ＞ 1 and P ＜ 0.05 was judged to be a risk factor.Results A total of 3090 cases information of patients with HFRS and 6018 healthy controls were collected.Of the 3090 cases of HFRS patients,sixty vaccinated patients showed atypical clinical manifestations and they were mild or moderate cases and no deaths.A total of 3030 nonvaccinated patients had obvious clinical symptoms.Severe or critical cases accounted for 39.07％(1184/3030)and 60 patients died and the fatality rate was 1.98％ (60/3030).The results of single factor Logistic regression analysis showed that of the 16 factors analyzed,the difference of 11 risk factors between the case group and the control group was statistically significant.Multivariate Logistic regression analysis showed that of the 11 factors,the difference of six factors between the case group and the control group was statistically significant.The main risk factors of suffering HFRS in the order were:exposure to rat pollutants,living in the affected areas,sitting or lying on grass fields in the affected areas,working in the affected areas,house rat infestation,and domesticated cats or dogs(OR =6.826,5.764,4.882,4.857,3.126 and 2.875; P ＜ 0.01 or P ＜ 0.05).Conclusions Vaccines are very useful in the sense that vaccinated HFRS patients tend to have mild symptoms and good prognosis.Health education in this area should be focused on the six risk factors in order to prevent the spreading of HFRS.