Updated clinical and glycomic features of mannosyl-oligosaccharide glucosidase deficiency:Two case reports

BACKGROUND Mannosyl-oligosaccharide glucosidase(MOGS)deficiency is an extremely rare type of congenital disorder of glycosylation(CDG),with only 12 reported cases.Its clinical,genetic,and glycomic features are still expanding.Our aim is to update the novel clinical and glycosylation features of 2 previously reported patients with MOGS-CDG.CASE SUMMARY We collected comprehensive clinical information,and conducted the immunoglobulin G1 glycosylation assay using nano-electrospray ionization source quadruple time-of-flight mass spectrometry.Novel dysmorphic features included an enlarged tongue,forwardly rotated earlobes,a birth mark,overlapped toes,and abnormal fat distribution.Novel imaging findings included pericardial effusion,a deep interarytenoid groove,mild congenital subglottic stenosis,and laryngomalacia.Novel laboratory findings included peripheral leukocytosis with neutrophil predominance,elevated C-reactive protein and creatine kinase,dyslipidemia,coagulopathy,complement 3 and complement 4 deficiencies,decreased proportions of T lymphocytes and natural killer cells,and increased serum interleukin 6.Glycosylation studies showed a significant increase of hypermannosylated glycopeptides(Glc3Man7GlcNAc2/N2H10 and Man5GlcNAc2/N2H5)and hypersialylated glycopeptides.A compensatory glycosylation pathway leading to an increase in Man5GlcNAc2/N2H5 was indicated with the glycosylation profile.CONCLUSION We confirmed abnormal glycomics in 1 patient,expanding the clinical and glycomic spectrum of MOGS-CDG.We also postulated a compensatory glycosylation pathway,leading to a possible serum biomarker for future diagnosis.

Isolation and screening of glucosidase inhibitors from Chinese medicines

Objective: To search for glucosidase inhibitors from Chinese medicines. Methods: Six kinds of widely-used Chinese medicines with the activity of decreasing blood glucose were prepared by the process of boiling, condensing, precipitating, exchanging with resins and rinsing. In vitro glucosidase inhibitory activities were examined by photometric bioassay derived from rats, yeast and almond of all the Chinese medicine extracts. Diabetic ICR mice models were established by intraperitoneal injection of STZ (200 mg/kg). To investigate the in vivo effect of lowering blood glucose, the mouse blood glucose level was assayed at 30 min after being given 2.5 g/kg starch and acarbose or varied concentrations of different constituents of some Chinese medicines by stomach tube. Results: The constituents of Sangye, Sangzhi, Sangbaipi, Dihuang and Yuzhu showed potent inhibitory activities against glucosidase. Furthermore, the first kind of constituents was proved to be beneficial in reducing blood glucose by in vivo glucose tolerance experiments. Conclusion: The constituents of Chinese medicines with reducing blood glucose effect have been discovered, thus providing a clue to novel drugs.

Requirement of a Homolog of Glucosidase Ⅱ β-Subunit for EFR-Mediated Defense Signaling in Arabidopsis thaliana

EFR is a plasma-membrane resident receptor responsible for recognition of microbial elongation factorTu （EF-Tu） and thus triggering plant innate immunity to fend off phytopathogens. Functional EFR must be subject to the endoplasmic reticulum quality control （ERQC） machinery for the correct folding and proper assembly in order to reach its final destination. Genetic studies have demonstrated that ERD2b, a counterpart of the yeast or mammalian HDEL receptor ERD2 for retaining proteins in the endoplasmic reticulum （ER） lumen, is required for EFR function in plants （Li et al., 2009）. In this study, we characterized the Arabidopsis glucosidase Ⅱ β--subunit via the H DEL motif against the non-redundant protein database. Data mining also revealed that the glucosidase Ⅱ β--subunit gene has a highly similar expression pattern to ERD2b and the other known ERQC components involved in EFR biogenesis. Importantly, the T-DNA insertion lines of the glucosidase Ⅱ β-subunit gene showed that EFR-controlled responses were substantially reduced or completely blocked in these mutants. The responses include seedling growth inhibition, induction of marker genes, MAP kinase activation, and callose deposition, trigged by peptide elf18, a full mimic of E F-Tu. Taken together, ourdata indicate a requirement of the glucosidase Ⅱ β-subunitfor EFR function.

High production of β-glucosidase from a marine Aspergillus niger immobilized on towel gourd vegetable sponges

To produce β-glucosidase by consecutive batch fermentation, a marine Aspergillus niger was immobilized on a natural carrier, towel gourd vegetable sponges. The immobilized mycelia were 0.15 g/g carrier with the immobilized biomass percentage of over 95%. The immobilized mycelia possessed the long durability（22.5 days）. The maximum production occurred 1.5 day earlier by the immobilized mycelia than by the free mycelia. β-Glucosidase production of five consecutive batches was over 110 U/m L. At high salinity,the activity and stability of β-glucosidase from the marine A. niger increased remarkable. Immobilizing the marine A. niger on the novel natural carrier achieved the efficient production of β-glucosidase.

Inhibitory Activities on α-Glucosidase and Composition Analysis of Different Fractions of Ethanol Extracts from Actinidia chinensis Radix

Tlie inhibitory actions of ethyl acetate,n-butanol and water fractions from Actinidia chinensis Radix (A. chinensis Radix) ethanol extracts on glucosidase were investigated. We found that the inhibitory activity of different ethanol extract fractions on α-glucosidase is concentration-dependent. At the same concentration, the ethyl acetate and n-butaiiol fractions showed higher inliibitory activities on gglucosidase in comparison witli acarbose. We then determined the chemical constituents of tlie ethyl acetate and n-butanol fractions through ultra performance liquid cliromatography-triple-time of flight/mass spectrometry(UPLC-Triple-TOF/MS). Based on the mass spectrometry analysis and the databases Scitinder and Reaxys, 21 compoiinds(A1-A21) were identified from the ethyl acetate fraction. In addition, 11 of those compounds(A2, A3, A5, A6, A8, A10, A11, A13, A15, A19, A20) were identified for the first time from A. chinensis Radix. We also identified 14 compounds(A22-A32 and A4-A6) from tlie w-butanol fraction and compound A29 is a new natural product. From the n-butanol fraction, 7 additional compounds (A22, A25, A27, A28, A30-A32) were identified from A. chinensis Radix for the first time.

GLUCOSIDASE INHIBITORY ACTIVITY AND HPLC ANALYSIS OF THE FLAVONOIDS OBTAINED FROM IMPATIENS BALSAMINA L. OF DIFFERENT REGIONS

A simple and efficient HPLC method was developed for quality analysis of flavonoids from Impatiens balsamina L.(IBL)flowers,which were collected from Xingjiang,Anhui,Henan,and Hubei provinces in China.The flavonoids substances in IBL were determined by HPLC through methyl alcohol ultrasonic extraction.A mixture of

Characteristics of Cellulases from Anoplophora glabripennis Motsch (Coleoptera: Cerambycidae)

The property of major cellulases from the guts of Anoplophora glabripennis larvae have been characterized. The optimal temperatures of both β 1,4 glucosidase (β glucosidase) and endo β 1,4 endoglucanase (endoglucanase, Cx) are 40℃. The β glucosidase was optimally active at pH 4\^8, while the optimal activity of the endoglucanase occurred at pH 4 4 5 6 The endoglucanase was active with a wide range of pH and temperature, the levels of activity from 25℃ to 50℃ were more than 80%, and the activity remained 60% between pH 3 2 and pH 7 2. The endoglucanase exhibited higher thermal stability than β glucosidase. Both enzymes lose their activities by heat treatment at 60℃. Two isozymes of endoglucanase were detected in sodium carboxymethylcellulose polyacrymide gels (CMC gel) by chemical colorization, and purified by elution from the gel slices. The molecular weights of the two isozymes were estimated as 26kD and 39kD respectively. Moreover molecular characteristics of the two isozymes are currently underway.

Effects of tripchlorolide on the epididymides and testes of rats

Aim: To further evaluate the antifertility effects of tripchorolide, a derivative of triptolide produced at the extraction pro-cedure of Tripterygium wilfordii Hook. f., in male rats and to investigate its sites and possible mechanisms of action.Methods: In male rats, tripchlorolide was given by oral garage at a dose of 50 ug.kg～(-l).d～(-1) for 5 weeks, fertility wasassessed by mating tests, and biochemical indices and light microscopic observation of the epididymides and testes werealso performed. Results: Administration of tripchlorolide at 50 ugg.kg～(-l)-d～(-1) for 3 weeks did not influence the fertilityof male rats, but 5-week treatment rendered the rats infertile. The density and motility of spermatozoa collected fromcauda epididymides were reduced significantly. The epididymal weights, as well as the L-carnitine concentration and α-glucosidase content in the epididymal fluid were decreasd. There were no significant differences in α-glucosidase andacid phosphatase (ACP) in caput epididymal homogenates between the control and the experimental rats. Obvious mor-phological changes were observed in the epididymal spermatozoa, mainly including head and tail separation or acrosomecurving. Sloughed spermatids were found in the seminifeous and epididymal tubules. In tesficular homogenates,tripchlorolide had no influence on the lactate dehydrogenase-C_4 (LDH-C_4) and hyaluronidase activities. No apparentlesions were observed in the seminiferous and epididymal epithelium. Conclusion: At the dose level employed,tripchlorolide has a significant effect on the fertility in male rats and the primary sites of action may be spermatids and tes-ticular and epididymal spermatozoa. (Asian J Androl 1999 Sep ; 1: 121 - 125)

血清AFU、LAP和GGT联合检测在早期原发性肝癌诊断中的应用与评价

我国是肝炎发病大国,肝癌发病率也呈逐年上升的趋势,是临床常见的疾病。据统计,每年大约有11万人因肝癌死亡。肝癌一般起病比较隐蔽,早期无典型的临床症状,临床上一旦确诊一般属于中、晚期。因此,如何早期诊断肝癌具有重要的意义。

Antidiabetic potential of methanol extracts from leaves of Piper umbellatum L. and Persea americana Mill.

Objective: To determine inhibitory activity of methanolic leaf extract of Piper umbellatum and Persea americana(P. americana)(traditionally used in Cameroon against diabetes) on β-glucosidase, β-glucosidase, maltase-glucoamylase, aldose reductase and aldehyde reductase activities, enzymes involved in starch digestion or diabetic complications. Methods: The methanol extracts from Piper umbellatum and P. americana were prepared by maceration. To assess relative efficacy of these extracts, the determination of concentrations that were needed to inhibit 50% of enzyme activity was done, whereas, gas chromatography-mass spectrum was used to identify components from extracts that may be responsible for the activities. Results:The tested extracts strongly inhibited β-glucosidase, maltase-glucoamylase, aldose reductase and aldehyde reductase activities with IC50 ranging from(1.07 ± 0.03) to(31.77 ± 1.17) μg/mL.Among the tested extracts, P. americana was the most active against sensitive enzymes(IC50 of1.07 ± 0.03 to 15.63 ± 1.23). But, none of the extracts showed interesting inhibitory effect against β-glucosidase as their percentage inhibitions were less than 16%. From gas chromatographymass spectrum analysis, 10 and 8 compounds were identified in Piper umbellatum and P.americana extracts respectively, using NIST library 2014. Conclusions: Results of this study provide the scientific credential for a prospective usage of these plants to treat diabetes.

Renal hemodynamics change and renal dysfunction during liver cirrhosis

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Digestive enzymes inhibition, antioxidant and antiglycation activities of phenolic compounds from jabuticaba (Plinia cauliflora) peel

In this study, jabuticaba peel purified phenolic extracts were studied regarding their potential inhibitory effects on digestive enzymes linked to diabetes mellitus (DM) and obesity, inhibitory activities towards glycation of bovine serum albumin (BSA), and antioxidant activities. The impact of in vitro gastrointestinal digestion (GID) on the stability of anthocyanins and bioactivities was also evaluated. The analyzed extracts exhibited significant inhibitory activities of α-glucosidase (IC50: 0.47–5.20 mg/mL), α-amylase (IC50: 0.45–1.88 mg/mL), and lipase (IC50: 13.52–111.51 mg/mL). The extracts also inhibited the glycation of BSA (IC50: 0.23–7.07 mg/mL) and showed relevant antioxidant activities. The correlation analysis indicated anthocyanins as one of the main contributing compounds to the bioactivities studied. Cyanidin-3-glucoside was the major anthocyanin identified in the extracts by HPLC analysis. After GID, there was a decrease in the total anthocyanin content (72.77% ± 1.67%) and, consequently, a reduction in the inhibitory activities of carbohydrate digestive enzymes and BSA glycation. This research provided scientific data about the functional properties of jabuticaba peel extracts, which may be a source of dietary compounds with hypoglycaemic, antiglycation, and antioxidant effects.

Phoenix dactylifera L.seed protein hydrolysates as a potential source of peptides with antidiabetic and anti-hypercholesterolemic properties:An in vitro study

This study aimed to explore the effect of enzymatic hydrolysis conditions(enzyme types and hydrolysis time)of date seed protein hydrolysates(DSPH)on in-vitro inhibition of molecular markers related with diabetic and hypercholesteremia.The DSPH was prepared using alcalase,bromelain,papain,and protease at different hydrolysis time(HT)(2,4 and 6 h).Higher degree of hydrolysis was observed for papain at 6 h HT.The results showed that alcalase and bromelain generated DSPHs at 6 h HT greatly improved the pancreatic lipase inhibition activity while the cholesterol esterase inhibition activity was greatly enhanced with alcalase and protease at 6 h HT and papain generated DSPHs at 4 h HT.Bromelain generated DSPH displayed the highest inhibition of dipeptidyl peptidase-IV(DPP-IV)and amylase at 6 and 2 h hydrolysis time,respectively.All the enzymes at 2 h HT except that of papain showed the highestα-glucosidase inhibitory activities.These results revealed that the DSPH displayed enhanced inhibitory activities towards molecular markers related with diabetes and obesity and thus have promising potential to be used as health-promoting ingredients in the fabrication of functional foods.

Pompe病的研究进展

Pompe＇s病又称为Ⅱ型糖原累积病（glycogen storage disease,typeⅡ,GSDⅡ）,由荷兰病理学家Pompe于1932年首先提出〔1〕,是一种由于先天性酸性α-葡萄糖苷（acidα-glucosidase,GAA）缺陷所导致的常染色体隐性遗传性代谢性疾病。近年来,特别是2006年美国FDA批准Myozyme作为Pompe＇s病的酶替代治疗（enzyme replacement therapy,ERT）药物以来,对GSDII的自然过程、Myozyme的疗效、免疫反应、病理、新生儿筛查、运动规律、陪伴治疗及基因治疗等方面取得了重要的进展，现就近年来研究进展综述如下。

The effects of simulated nitrogen deposition on extracellular enzyme activities of litter and soil among different-aged stands of larch

Aims Nitrogen(N)addition could affect the rate of forest litter and soil organic matter decomposition by regulating extracellular enzyme activity(EEa).The impact of N addition on EEa may differ across different age stands with different organic matter quality.We were interested in whether the impact of N addition on EEa in litter and mineral soil during the growing season was dependent on stand age of a larch plantation in North China.Methods We added three levels of N(0,20 and 50 kg N ha^(−1) year^(−1))in three age stands(11,20 and 45 years old)of Larix principis-rupprech-tii plantation in North China.We measured potential activities of β-1,4-glucosidase(b),cellobiohydrolase(Cb),β-1,4-N-acetyl-glucosaminidase(Nag)and phenol oxidase(Po)in litter(organic horizon)and mineral soil(0-10 cm)during the second growing sea-son after N amendment.We also measured C and N concentrations,microbial biomass C and N,and KCl-extractable ammonium and nitrate in both litter and mineral soil.Important Findings We observed unimodal patterns of EEa during the growing season in all three stands,consistent with the seasonal variations of soil temperature.stand age had a strong effect on EEa in both litter and mineral soil,and this effect differed between litter and mineral soil as well as between different enzymes.N addition did not significantly affect the activities of b or Cb but significantly suppressed the activity of Nag in litter.We also found stand age-specific responses of Po activity to N addition in both litter and mineral soil.N addition suppressed Po activity of the high C:N ratio litters in 20-and 45-year-old stands but had no significant effect on Po activity of the low C:N ratio litter in 11-year-old stand.moreover,N addition inhibited Po activity of the high C:N ratio soil in 20-year-old stand but had no significant impact on Po activity of the low C:N ratio soils in 11-and 45-year-old stands.overall,stand age had a greater effect on EEa in litter and mineral soil compared to 2 years of N addition.moreover,the effect of N addition on Po activity is stand age dependent,which may affect the long-term soil carbon storage in this forest.

RP-HPLC Determination of 1,3-Dideoxygalactonojirimycin in Bombycis Faeces

Objective To establish a simple and rapid method for the determination of 1,3-dideoxygalactonojirimycin in Bombycis Faeces,a potent glucosidase inihibitor,by HPLC.Methods A RP-HPLC method with fluorescence detection has been developed.Results The HPLC method developed in this research has a good reliability including accuracy and precision.The detection limit was less than 72 ng.Conclusion This method is sufficiently sensitive for determining 1,3-dideoxygalactonojirimycin in Bombycis Faeces and other related products.