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 evaluate cognitive function of chronic alcoholism patients.Methods The Wechsler Adult Intelligence Scale-Revise in China(WAIS-RC) and WMS-RC,including intelligence quotient,logical Memory.visual memory and stroop test,were investigated and P300 component of auditory event-related potential (ERP,P300),especially the subcomponents P3a and P3b were examined in 100 chronic alcoholism patients,and 70 healthv people matched in age,sex and educational level as controls.Results Compared with controls,the score of Verbal intelligence quotient(VIQ),Performance intelligence quotient (PIQ),Full-scale intelligence quotient (FIQ),logical memory,visual memory (immediate and delayed),and stroop test were significant lower (P <0.05.P＜O.01)in chronic alcoholism patients,but the latencies of P3a and P3b wave were significantly prolonged.the amplitude of P3a and P3b wave were significantly decreased in chronic alcoholism patients.Conclusion Chronic alcoholism patients have cognitive function impairment,and P300 is a more sensitive to discover the impairment of the cognitive function.

As a treasure resource of novel drug lead compounds, how to rapidly and high-efficiently screen and isolate active components from natural products is critical. Thanks to its high resolution, high automation and flexible integration, online two-dimensional liquid chromatography has great potential for screening active ingredients from complex matrices by integrating a highly specific bio-recognition process into a two-dimensional liquid chromatography system before, on or after the column separation. This review comprehensively summarized recent developments, applications and shortcomings of online two-dimensional liquid chromatography for natural product screening from different integration modes, including pre-column, on-column and post-column screening methods.

Objective To evaluate the laparoscopic Palomo procedure in the management of varicocele in adolescent by assessment of the postoperative testicular growth and the changes of the biggest vessel diameter of varix. Methods From July 1999 to March 2004, laparoscopic high mass - ligation of testicular vessels according to Palomo procedure was performed on 31 boys who all presented with left varicocele Before and after operation at 1 month,3 and 6 months, the biggest vessel diameter of varicocele was measured by ultrasonography, testicular blood supply was evaluated by color Doppler flow imaging(CDFI) ,and fastis was recorded in length, width and thickness by ultrasonographic measurement, which volume was calculated by the formula V =?/6 ? length ? width ? thickness. Also the ratio of left to right testicular volume was calculated, and the left testicle was thought atrophy if the ratio was below 75% The biggest vessel diameter of varix, testicular blood supply, and the postoperative testicular increasing volume were compared before and after operation. Results The biggest vessel diameter of varicocele significantly reduced 1 month after operation(P

Background and purpose:Recently, studies showed that non-steroidal anti-inlfammatory drugs (NSAIDs) could reduce the incidence of cancer. Whether ibuprofen could inhibit the growth of hepatocellular carcinoma cells had not been reported yet. In the current study, we investigated the effects of ibuprofen on hepatoma carcinoma BEL-7402 cells and the relevant mechanisms. Methods: Hepatocellular carcinoma BEL-7402 cells were randomly divided into 7 groups:the control group and the ibuprofen groups (0.1, 0.5, 1.0, 2.0, 3.0 and 4.0 mmol/L). The effect of ibu-profen on BEL-7402 HCC cells was measured by MTT method, the cell cycles were analyzed by flow cytometry (FCM), cell vitality and apoptosis were determined by cell analyzer. PCNA, Cyclin D1, Bcl-2 and COX-2 protein levels were examined by Western blot, and the expressions of prostaglandin E2 (PGE2) were measured by ELISA. Results:After the exposure to ibuprofen, the suppression ratio of BEL-7402 cells was increased (P<0.05). BEL-7402 cell vitality was decreased by degrees significantly (P<0.05), early apoptosis of BEL-7402 cells was increased (P<0.05), and the G0/Gl phase ratio was increased significantly compared with control group (P<0.05). Ibuprofen effectively decreased PCNA, Cyclin D1, Bcl-2 and COX-2 expressions in BEL-7402 cells (P<0.05), and decreased PGE2 protein expression in cell culture supernatants sig-nificantly (P<0.05). Conclusion:Ibuprofen is effective for inhibiting the proliferations, increasing apoptosis and blocking cell cycles of BEL-7402 HCC cells. The anti-tumor mechanisms of ibuprofen may be related with the inhibition of COX-2 and PGE2 expressions.

In the face of escalating problems with pathogen control, the development of proper formulations of existing antibiotics is as important as the development of novel antibiotics. Daptomycin is a lipopeptide antibiotic with potent activity against Gram-positive bacteria. Currently, only injectable solution of daptomycin has been approved for clinical use. In the present study, the formulation of PEGylated liposomal daptomycin (PLD) was prepared and optimized, and its efficacy against methicillin-resistant Staphylococcus aureus (MRSA252) strains was investigated. The obtained PLD had a mean vesicle diameter of (111.5 +/- 15.4) nm and a mean percent drug loading of (5.81 +/- 0.19) % with high storage stability. Potent activity of PLD against MRSA was demonstrated in vitro with a more sustained effect than that of conventional liposomal daptomycin and daptomycin solution. In addition, intravenous administration of a single dose (equal to human use) of PLD significantly increased the survival of mice in a MRSA252 systemic infection model compared with other formulations. Drug distribution in the lung was significantly enhanced following administration of PLD, and no measurable tissue lesions or pathological changes were detected during PLD treatment. Taken together, PEGylated liposomes loaded with daptomycin may represent a promising approach to reduce MRSA252 infections, especially those involving bloodstream dissemination, such as hematogenous pulmonary infection.
Animals ; Anti-Bacterial Agents ; pharmacology ; Daptomycin ; pharmacology ; Liposomes ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Mice ; Staphylococcal Infections ; drug therapy

Objective To investigate the effect of fluoride on the expression of a subunit of calcium/calmodulin-dependent protein kinase- Ⅱ (α-CaMK Ⅱ ) at both mRNA and protein levels in human neuroblastoma cells were cultured in DMEM with final concentrations of NaF 0(control) ,0.05,0.50,2.00,5.00 mmol/L, respectively, for 48 hours. Then quantitative RT-PCR and Western blot were performed to detect the expression level of α-CaMK Ⅱ P1 (postnatal day 1) pups together with their mothers were randomly divided into three groups. Lactating rats were given drinking water containing NaF at concentrations 0(control) ,2,3 mmol/L. And pups were exposed to NaF through milk. In each group, 8 pups were sacrificed on day 14 after birth. In post-weaning period, another 8 pups in each group were given drinking water with the same dose of fluoride as their mother's 21 day after birth. After then, these pups were killed on day 28, and hippocampus was dissected immediately and Western blot was conducted mRNA and protein levels were decreased. When NaF concentrations were 0,0.05,0.50,2.00,5.00 mmol/L, the mRNA relative ratios of α-CaMKⅡ in SY5Y cells were 1.00 ± 0.00,0.77 ± 0.18,0.40 ± 0.11,0.22 ± 0.06 and 0.15 ± 0.03, and protein levels of α-CaMK Ⅱ were 100.00 ± 0.00,76.17 ± 2.08,59.16 ± 2.12,48.52 ± 2.71 and 43.51 ± 2.57, any mmol/L group, hippocampus α-CaMK Ⅱ protein levels on day 14 and 28(75.02 ± 2.88,73.83 ± 3.88 and 81.00 ± 2.54,45.70 ± 2.34) were significantly lower than that of control groups(100.00 ± 0.00,100.00 ± 0.00, all P < 0.01). In 3 mmol/L group, hippocampus α-CaMKⅡ protein level on day 28 was lower than that of 2 mmol/L group (P < 0.01). Conclusion Fluoride can decrease mRNA and protein levels of α-CaMK Ⅱ in nerve cells and hippocampus, which may be one of the mechanisms of learning and memory impairment by fluorosis.

Objective To evaluate accuracy and consistency of the coagulation convention that involves PT, INR, FIB, APTT on two coagulation analysators(ACL-9000 and ACL-200, Beckman-Coulter Corporation, US). Methods The repeatability test was used to assess the precision, the accuracy and consistency of the coagulation convention on two coagulation analysators were evaluated by comparison and analysis. Results The repeatability of related test results based on ACL-9000 coagulation analysator was better that CV value less than 5%, but the repeatability of abnormal specimens FIB based on ACL-200 was poor that CV value of 6.6%. Through comparing ACL-9000 analyzer, the test results of ACL- 200 coagulation analysator has good correlation with it, correlation coefficient ?﹥0.975. Conclusion In the condition of the excellent precision and stability, we can also find system errors existed in the analyzer on time by verificating. Through being adjusted and calibrated, the examination department can deliver the results of accuracy consistency.

Objective To study the impact and mechanisms of type 2 diabetes mellitus( T2DM) on mobi-lization of hematopoietic stem cells of multiple myeloma( MM) .Methods 48 patients diagnosed with MM whose hematopoietic stem cells were collected entered the study, including 15 cases with DM, and 33 cases without DM.29 were males and 19 were females, with 55.2 as the median age, ranging from 41 to 63 years.We ob-served numbers of CD34 +cells collected, CD11c positive cells and CD206 positive cells in bone marrow, then the ratio of CD11c/CD206 was considered as the ratio of M1 macrophages/M2 macrophages, in the end we explored the relevance of macrophage polarization status with the number of CD34 +cells collected.Results ①The num-ber of CD34 +cells collected in MM patients was(4.63 ±1.75) ×106/kg, while the number in MM patients with DM was(3.12 ±1.84) ×106/kg.The number of CD34 +in MM group was significantly higher than that in the MM with DM group(P<0.05).②CD11c/CD206 cell ratio in MM patients was 1.42 ±0.16, while the ratio was 3.08 ±0.37 in MM patients with DM, indicating that MΦM1 polarization occurred in the MM with DM group.③CD11c/CD206 cell ratio was 2.87 ±0.42 in 6 patients whose stem cell collection number was less than 2 ×106/kg, the ratio was 1.91 ±0.28 in 14 patients whose stem cell collection number was between 2 ×106/kg and 5 × 106/kg, and the ratio was 1.48 ±0.25 in 8 patients whose stem cell collection number was more than 5 ×106/kg.There were significant differences in these 3 groups( P<0.05) , indicating that macrophage polarization and the number of stem cell collection was relevant.Conclusions DM is unfavorable factor in stem cell mobilization and collection of multiple myeloma, it may be related to macrophages M1 polarization induced by diabetes.

Objective To explore the relationship between the gait behavior changes and cognitive function in APP/PS1 transgenic mice .Methods 16 APP/PS1 transgenic mice were divided into model group and Huperzine A group, C57/BL6J mice with the same age were chosed as control group .After a 150 days consecutive treatment , Morris water maze(MWM) was used to detect the learning and memory ability and Gait analysis system (GAS-2) was used to detect the gait behavior after the treatment when the mice were 8-month-old.Results The escape latency of the model group was significantly higher than that of the control group ( P <0.05 ) , the time spended in the target quadrant , swimming distance in the target quadrant significantly lower than that of the control group ( P <0.05 ) , the first time passing through the platform prolonged significantly than the control group (P <0.05), and the number of passing though the platform reduced significantly than the control group (P <0.05).In the gait behavior experiments , compared with the control group, the average walking speed of the model group reduced significantly (P <0.05), the average walking cycle, the absolute average body angle and lateral movement increased significantly (P <0.05);The percentage of support time in a walking cycle of the left and right foot increased significantly (P <0.05).Accordingly, the percentage of swing time in a walking cycle of the left and right foot reduced significantly (P <0.05).The propulsion index of the left hand , right hand, right foot increased significantly ( P <0.05), and then the braking index of the above three feet decreased significantly ( P <0.05) .Huperzine A can improve the cognitive function , rectify the changes in the gait behavior .The two behavioral relevance shows that cognitive function and the front two feet braking , propulsion index have a high correlation index (correlation coefficients were -0.433, -0.379, P values were 0.039,0.079), the others were not. Conclusion APP/PS1 transgenic mice of 8-months-old have a remarkable impairment of learning and memory ability and disorder of gait behavior , and these two behaviors have a correlation in some extent .