Congenital disorders of glycosylation(CDG) are a rapidly growing group of genetic diseases that are due to defects in the synthesis of glycans and in the attachment of glycans to other compounds. Most CDG are multisystemic diseases often involving severe psychomotor retardation. The CDG causing sialic acid deficiency of N-glycans can be diagnosed by isoelectric focusing of serum transferrin. Glycan structural analysis, yeast genetics and knockout animal models are essential tools in the elucidation of novel CDG. In this review, we focus on the current knowledge of the pathogenesis and identification of nine primary glycosylation diseases.

AIM: To investigate the effects of uric acid sodium salt (UANa) as adjuvant on humoral and cellular immune response in BALB/c mice. METHODS: BALB/c mice were immunized with trichosanthin (TCS) as antigen together with UANa suspension as adjuvant. The antibody titers of IgG were detected by enzyme-linked immunosorbent assay. Dendritic cells (DC) were induced in vitro, the phenotypes of DC were analyzed by flow cytometry and the effect of UANa on DC maturity was evaluated. A delayed-type hypersensitivity (DTH) model was used to analyze the effect of UANa on cellular immune responses in vivo. The in vitro proliferation of lymphocytes was determined by ConA stimulation. RESULTS: Freunds adjuvant greatly enhanced the antibody response of mice to TCS, while UANa adjuvant failed to promote the antibody response but significantly reduced the antibody response as compared to TCS only. No effect of UANa on the expression of CD11c and CD83 in DC was observed by flow cytometry analysis. However, UANa significantly enhanced the expression of MHC II molecule. In the DTH model, UANa enhanced the degree of allergen-induced ear swelling and promoted the ability of lymphocyte proliferation in vitro. CONCLUSION: UANa suspension as adjuvant significantly enhances the cellular immune response but inhibits the humoral immune response to a certain degree, suggesting that UANa has potential application in the vaccine research.

AIM:To investigate the effects of uric acid sodium salt (UANa) as adjuvant on humoral and cellular immune response in BALB/c mice. METHODS:BALB/c mice were immunized with trichosanthin (TCS) as antigen together with UANa suspension as adjuvant. The antibody titers of IgG were detected by enzyme-linked immunosorbent assay. Dendritic cells (DC) were induced in vitro,the phenotypes of DC were analyzed by flow cytometry and the effect of UANa on DC maturity was evaluated. A delayed-type hypersensitivity (DTH) model was used to analyze the effect of UANa on cellular immune responses in vivo. The in vitro proliferation of lymphocytes was determined by ConA stimulation. RESULTS:Freund's adjuvant greatly enhanced the antibody response of mice to TCS,while UANa adjuvant failed to promote the antibody response but significantly reduced the antibody response as compared to TCS only. No effect of UANa on the expression of CD11c and CD83 in DC was observed by flow cytometry analysis. However,UANa significantly enhanced the expression of MHC II molecule. In the DTH model,UANa enhanced the degree of allergen-induced ear swelling and promoted the ability of lymphocyte proliferation in vitro. CONCLUSION:UANa suspension as adjuvant significantly enhances the cellular immune response but inhibits the humoral immune response to a certain degree,suggesting that UANa has potential application in the vaccine research.

From the n-butanol extract of the leaves of Callicarpa nudiflora,thirteen compounds were isolated by repeated column chromatography with silica gel,C_(18) and Sephadex LH-20.Their structures were identified by spectroscopic (~1H,~(13)C NMR and so on) and chemical methods as luteolin (1),luteolin-7-O-β-D-glucopyran coside (2),apigenin (3),5,7,4'-trihydroxy-3'-methoxyflavone (4),luteolin-3'-O-β-D-glucopyranoside (5),api genin-7-O-β-D-glucopyranoside (6),luteolin-7-O-(6-trans-caffeoyl)-β-D-glucopyranoside (7),luteolin-7-O-(6-trans-feruloyl)-β-D-glucopyranoside (8),arjunglucoside I(9),luteolin-7-O-(6-p-coumaryl)-β-D-glucopyranoside (10),forsythoside B (11),isomartynoside (12),deacylisomartynoside B (13).Among them,compound 11 was isolated from this plant for the first time and compounds 4,7-10,12-13 were isolated from this genus for the first time.

Objective To evaluate the response of the lung tumor xenografts in nude mice to antiangiogenic treatment from perspectives of anatomic,vessel function,cellular and molecular level using the multimodality imaging techniques including optical imaging,dynamic contrast enhanced MRI(DCE-MRI)and diffusion weighted imaging(DWI).Methods The green fluorescent protein(GFP)was transplanted labeled using GFP-expressing NCI-H460 cells.After the transfection of GFP,NCI-H460 cells were implanted subcutaneously into nude mice.Ten days after implantion,12 nude mice whose tumor xenografts grew to 0.5-1.0 cm in the maximum diameter were randomly divided into 2 groups,and injected with phosphate-buffered saline and recombinant human endostatin respectively.Then the nude mice in the two groups underwent optical imaging,DCE-MRI and DWI.The volumes,photon counts,the quantitative MR vessel functional parameters including volume transfer constant(Ktrans),rate constant(Kep),volume of extravascular extracellular space(Ve)and maximum area under the enhancement curve(iAUC),and apparent diffusion coefficient(ADC)values of the tumors were recorded.Then tumors were collected and observed using the transmission electron microscopy and pathology examination,including HE staining,microvessel density(MVD)and the expressions of vascular endothelial cell growth factor(VEGF).The Kep and VEGF expressions in experimental group and control group were compared with x2 text,and other values were compared with t test.The Pearson and Spearman test were used for analyzing the correlation of values in the two groups.Results Seven days after inoculation,the fluorescence signals were detected and grew with the growth of the tumors.On the 7 day after starting therapy,the photon counts of experimental group and control group were(2.51 ± 2.43)× 1010(photon/sec)and(5.77 ± 3.25)× 1010(photon/sec),respectively with no significant differences(t =1.964,P ＞0.05).Two sample t test showed that the tumor volumes in experimental group were smaller than those in control group[(365 ± 56)vs(987 ± 265)mm3,t =0.001,P ＜ 0.01].There was a positive correlation(r =0.673,P ＜ 0.05)between the photon counts and the volumes of the tumors.The mean Ktrans,Kep,Ve and iAUC of experimental group were:(0.055 ±0.012)min-1,0.335(0.184—0.894)min-1,0.297 ± 0.041 and 7.334 ± 3.930,and those for control group were:(0.117 ± 0.027)rin-1,0.417(0.324-1.736)min-1,0.326:±:0.062 and 13.280 ± 4.245.There were significant differences of Krans and iAUC(t =5.155,2.518,P ＜ 0.05)between experimental group and control group.And there was a positive correlation(r =0.715,P ＜ 0.0 1)between the values of iAUC and MVD,but not the expressions of VEGF(r =0.484,P ＞ 0.05).The values of ADC in experimental group were higher than that in control group[(791 ± 38)× 10-6 vs(737 ± 43)×10-6 mm2/s],and there were significant differences(t =-2.299,P ＜ 0.05).Two sample t test showed that the MVD in experimental group were lower than that in control group[(11.9 ± 4.8)vs(19.2 ±4.3)item/hpf,t =2.774,P ＜ 0.05].The VEGF expressions in experimental group were lower than that in control group(x2 =4.000,P ＞ 0.05).It was observed that some cells in experimental group had degenerated and apoptotic signs by the electron microscopy.Conclusions Evaluating the response of lung tumor xenografts to antiangiogenic treatment at anatomical,vessel functional,cellular and molecular level using the multimedality imagings is applicable.And it will be in favour of evaluating the therapeutic effect promptly.

This study was aimed to optimize the alcohol precipitation techniques for honeysuckle extract in order to standardize the production of honeysuckle extract of Qingkailing Injection and reduce the differences among batches. The orthogonal test was applied in this study. The content of chlorogenic acid and galuteolin were taken as compre-hensive indicators. Multi-index comprehensive scoring method was used in the data analysis. Three influencing fac-tors, which were the fluid temperature, the stirring speed and the speed of adding alcohol, were optimized in the al-cohol precipitation techniques. The results showed that the optimal alcohol precipitation techniques were when the fluid temperature was 20℃, the stirring speed was 240 rpm and the speed of adding alcohol was one time of the ma-terial per minute. It was concluded that the optimized alcohol precipitation process was stable and feasible.

AIM: Valproic acid (VPA) is a histone deacetylase inhibitor and is believed to have anti-tumor activity. The present study aims to investigate the effect of VPA on the, apoptosis and cytokine synthesis of human peripheral lymphocytes. METHODS: The activation and cytokine synthesis in lymphocytes in whole blood stimulated with phorbol dibutyrate (PDB) and ionomycin were evaluated with flow cytometry after fluorescent staining. The mitochondrial membrane potential was examined using 3, 3-dihexyloxacarbocyanine iodide [DiOC6(3)]staining. RESULTS: VPA at low doses (1 and 5 mmol/L) promoted CD69 expression in activated lymphocytes, whereas it turned to inhibit the expression of CD69 at a high dose (25 mmol/L). Meanwhile, VPA at low doses increased the mitochondrial membrane potential, while a high dose of VPA decreased it in activated lymphocytes. Furthermore, interleukin-2 (IL-2) synthesis was enhanced by low doses of VPA but inhibited by a high dose. However, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) synthesis were dose-dependently enhanced by VPA as compared with those of PDB plus ionomycin-treated cells. CONCLUSION: VPA exerts biphasic effect on the further activation and apoptosis of human peripheral lymphocytes stimulated with mitogens and exhibits differential activity on the synthesis of several important cytokines in human lymphocytes.

Objective: To introduce a new surgical technique for the correction of congenital syndactyly without skin grafts. Methods: The technique consists of a dorsal double-wing flap created from the dorsal skin of the metacarpophalangeal joint to cover the newly released web space and zigzag incisions in the fingers, thus avoiding the use of skin grafts in this area. Results: From May 2010 to October 2016, 35 web spaces in 24 patients were treated using this technique. There were no complications such as haematoma, infection or flap necrosis. The average follow-up time was 54 months (range, 6 months to 60 months). One of the 35 webs developed web creep. No patients developed flexion contractures. All webs had good appearance with 45 degrees inclination from dorsal to palmar. All fingers had no obvious scar, and the flexion and abduction function were good. Conclusions The technique is simple, rapid, safe and easily performed and does not require the use of skin grafts.

Objective To study the characteristic of absorption of euphorbia factor L1 in intestine of rats, and to observe the effects of P-glycoprotein(P-gp)and multidrug resistance-associated protein(MRP2)on intestinal absorption of euphorbia factor L1 .Methods The contents of euphorbia factor L1 of intestinal perfusion fluid of duodenum, jejunum, ileum and colon in the rats in situ single-pass intestinal perfusion model were determined by using HPLC.The drug absorption rate constant( Ka ) and the apparent absorp-tion coefficient( Papp ) in four intestinal regions were calculated.Results Ka and Papp of euphorbia factor L1 at colon were the highest of the whole rat intestine.Significant increase of Ka and Papp showed at rat colon when co-perfused with verapamil hydrochloride, by contrast, decrease of Ka and Papp found when co-perfused with indomethacin.Conclusion We inferred that verapamil hydrochloride be the substrate of P-gp and that indomethacin be not the substrate of MRP2 .

Objective:To explore the feasibility of Ponseti method in treatment of secondary clubfoot in young children with Tethered Cord Syndrome(TCS).Methods:The clinical data of 53 young children with clubfeet treated with Ponseti method from March 2014 to March 2017 at Department of Pediatric Orthopedics, the Third Affiliated Hospital of Zhengzhou University were analyzed retrospectively. These patients were divided into TCS group and Idiopathic group according to the etiology. There were 19 patients (33 feet) in TCS group,with an mean age of 2.8 months(range:0.2 to 24.0 months), including 13 males and 6 females, 5 patients with unilateral clubfeet and 14 patients with bilateral clubfeet. There were 34 patients (45 feet) in idiopathic group, with an mean age of 3.1 months(range: 0.1 to 21.0 months), including 18 males and 16 females, 23 patients with unilateral clubfeet and 11 patients with bilateral clubfeet. All the children received casts correction according to Ponseti method, and were followed up at 3 weeks, 3 months, 6 months and every 6 months after the Achilles tendon tenotomy or the last cast correction. Complications were recorded and therapeutic effect was evaluated of these children by Dimeglio Scoring System and the International Clubfoot Study Group (ICFSG) at the last follow-up. Independent t test, Mann-Witney U test or χ 2 test were used to compare the indicators of the two groups. Results:The number of plaster fixation in TCS group was (6.1±2.0) times, and that of idiopathic group was (4.8±1.0) times( t=3.482, P<0.01).In TCS group, 22 feet treated with Achilles tendon transection and that of idiopathic group was 40 feet(χ 2=0.279, P=0.598). There were 18 cases recurrence in TCS group and 8 cases in Idiopathic group ( t=11.149, P<0.01). In TCS group, 16 cases (27 feet) completed the initial correction, the success rate was 60.6% (27/33), 3 cases (6 feet) could not correct the deformity after 9 to 10 times of plaster fixation, and then underwent soft tissue release.In idiopathic group, 34 cases (45 feet) achieved initial correction after Ponseti treatment(χ 2=6.488, P=0.011).At the last follow up, there were 5 cases (9 feet) in TCS group and 2 cases (2 feet) in idiopathic group underwent soft tissue release(χ 2=6.110, P=0.013). The classification grade of ICFSG score of the two groups without soft tissue release were (2.1±0.6) and (1.8±0.7), the difference was not statistically significant ( t=1.765, P=0.082). All the children had no skin ulceration, bedsores, skin allergy and other complications. Conclusion:Ponseti method is effective in the treatment of clubfoot secondary to TCS, and the functional recovery is similar to that of children with idiopathic clubfoot.