Search

Pomegranate peel-derived extracellular nanovesicles(PPENs) were isolated and purified by ultra-high speed centrifugation and sucrose density gradient centrifugation. Their morphology and structure were characterized. In vitro α-glucosidase inhibition assay and model test of insulin resistance(IR) in HepG2 cells showed that PPENs had good anti-diabetic activity. The IC_(50) value of α-glucosidase inhibition was(35.3±1.1) μg·mL~(-1), significantly better than the positive drug acarbose. At a concentration of 100 μg·mL~(-1), PPENs could increase the glucose absorption of IR cells significantly. Lipidome, proteome, and metabolite analysis of PPENs were performed using chromatography-mass spectrometry. MicroRNA(miRNA) sequences were identified, and target genes of miRNA were predicted. The analysis results indicated that PPENs contained abundant lipids and transport proteins, providing a material basis for the transportation and distribution of PPENs in tissue. Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis suggested that lipids and miRNAs may be the key components of PPENs to exert anti-diabetic activity.

Pompe disease, also called type II glycogen storage disease, is a rare autosomal recessive inherited disease caused by the storage of glycogen in lysosome due to acid α-glucosidase (GAA) deficiency, with the most severe conditions in the skeletal muscle, the myocardium, and the smooth muscle. Patients may have the manifestations of dyspnea and dyskinesia, with or without hypertrophic cardiomyopathy. GAA gene mutation has ethnic and regional differences, and new mutation sites are found with the advances in research. Gene analysis is the gold standard for the diagnosis of Pompe disease. Conventional methods, such as skin and muscle biopsies and dried blood spot test, have certain limitations for the diagnosis of this disease. In recent years, prenatal diagnosis and newborn screening play an important role in early diagnosis of this disease. Enzyme replacement therapy (ERT) has a satisfactory effect in the treatment of this disease, but it may lead to immune intolerance. New targeted gene therapy and modified ERT will be put into practice in the future. This article reviews the research advances in the diagnosis and treatment of Pompe disease.

Objective: To explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS)., Methods: We randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR., Results: The concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05)., Conclusion: Ornidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

In order to develop potent antidiabetic agents that have inhibitory effect to a-glucosidase, twelve β-acetamido ketone derivatives such as N-{[(substituted-4-oxo-thiochroman-3-yl)phenyl]-methyl}acetamide are designed and synthesized through one-pot Dakin-West reaction. Their chemical structures are confirmed by 1H NMR, 13C NMR, IR and HR-MS. In vitro α-glucosidase inhibition assays of compounds 4a-41 were carried out using glucose oxidase method. The result indicated that most of them possess inhibitory activity in vitro. Compound 4k showed the most potent inhibitory activity with 87.3% inhibition of α-glucosidase at the concentration of 5.39 mmol x L(-1). The structure-activity relationship of these β-acetamido ketone derivatives was discussed preliminarily. Moreover, the molecular docking method was used to study the interaction mode of compound 4k and α-glucosidase. Our results will be helpful for designing of α-glucosidase inhibitors in the future.

Objective: To summarize clinical features and diagnosis of Chinese infantile patients with glycogen storage disease type II (GSD II)., Method: Six infant patients with GSD II diagnosed from January 2012 to June 2014 in the Department of Pediatrics, Peking University First Hospital were enrolled into this study. Clinical information of the 6 patients, including clinical manifestation, blood biochemistry, chest X-ray, echocardiogram, electrocardiogram, acid alpha-glucosidase (GAA) activity and GAA gene mutation analysis by direct sequencing of polymerase chain reaction (PCR) product were reviewed., Result: Of the 6 patients, five were female and one was male, five of whom were classic infantile type while the other one was atypical. The age of onset ranged from birth to 3-month-old. All patients had varying degrees of generalized muscle weakness, hypotonia and development retardation or retrogression. Other common findings were feeding difficulties in two patients, tongue weakness in two patients, respiratory distress in four patients, macroglossia in one patient, and hepatomegaly in two patients. Left ventricular hypertrophy and cardiomegaly were obvious in all the six patients. All six patients were found to have a enlarged heart in physical examination, and three patients who underwent a chest X-ray examination had an enlarged heart shadow. Four patients who had an echocardiography were found to have myocardial hypertrophy. The electrocardiogram in three patients showed short PR intervals and high voltage. The creatine kinase (CK) levels were three to seven times elevated. The mildest elevated CK was 441 IU/L, and the highest CK level was 1 238 U/L. Assay of GAA enzyme activity in whole blood showed significantly reduced activity (1.3 nmol/ (spot·d) to 2 nmol/(spot·d)) in the patients tested. Gene sequencing in 4 patients showed 8 pathogenic mutations, including 6 missense mutations, one nonsense mutation and one frameshift mutation. The missense mutations were c.998C > A (p.Thr333Lys), c.1280T > C (p.Met427Thr), c.1760T > C (p.Leu587Pro), c.1924G > T (p.Val642Phe), c.2012T > A (p.Met671Lys) and c.2105G > A (p.Arg702His). The nonsense mutation was c2662G > T (p.Glu888X), and the frameshift mutation was c2812&#95;2813delTG (p.Cys938fs). The 5 classic infantile patients died at the age of 7 to 22 months. The atypical infantile patient was 2 years and five months old according to our latest follow up., Conclusion: Infantile GSD II had similar motor manifestations and cardiac involvements, blood biochemical test, imaging findings, enzyme assays, though there were slight differences. The probability of GSD II should be taken into consideration if an infant has both muscular disease and cardiac involvement.

The present work was aimed to study soil a-glucosidase and beta-glucosidase activities of and red soils based on fluorescence detection method combined with 96 microplates with TECAN Infinite 200 Multi-Mode Microplate Reader. We added biochar or straw (2.5 g air dry sample/50g air dry soil sample) into and red soils and the test was carried under fixed temperature and humidity condition (25 degrees C, 20% soil moisture content). The results showed that straw addition enhances soil alpha-glucosidase and beta-glucosidase activities, beta-glucosidase activity stimulated by rice straw treatment was higher than that of corn straw treatment, and activity still maintains strong after 40 days, accounting for increasing soil carbon transformation with straw inputting. Straw inputting increased soil nutrients contents and may promote microbial activity, which also lead to the increase oin enzyme Straw inputting increased soil nutrients contents and may promote microbial activity, which also lead to the increase oin enzyme activities. Different effects of straw kinds may be related to material source that needs further research. However, biochar inputting has little effect on soil alpha-glucosidase and beta-glucosidase activity. Biochar contains less available nutrients than straw and have degradation-resistant characteristics. Compared with the conventional spectrophotometric method, fluorescence microplate method is more sensitive to soil enzyme activities in suspension liquid, which can be used in a large number of samples. In brief, fluorescence microplate method is fast, accurate, and simple to determine soil enzymes activities.

Objective: To investigate the effects of Xanthium sibiricum Patrin ex Widder water extracts (CEW) on α-glucosidase activity (AG) and blood sugar in mice., Methods: The inhibition of AG by CEW was studied with enzyme-inhibitor screening external model with acarbose as control drug. The normal mice were administrated by gavage with 40.0g*kg(-1), or 10.0 g*kg(-1) of CEW, 0.375 g*kg(-1) of acarbose, and 0.3ml of normal saline, respectively in successive 5 days; then the animals were loaded with 2.0 g*kg(-1) of glucose, 4.0 g*kg(-1) of sucrose, and 2.0 g*kg(-1) of starch and blood sugar levels were measured within 15, 30, 60, and 120min. Diabetes was induced by injection of streptozotocin (STZ) in mice, then 40.0 g*kg(-1), or 10.0 g*kg(-1) of CEW was given to diabetic mice in successive 2 weeks and 4 weeks, then the blood sugar levels were measured., Results: In the enzyme inhibition test, when the concentration of CEW was between 0.3125 g*L(-1)-10.00 g*L(-1), the inhibition rate was 55.42%-92.73% when the concentration of acarbose was 1.5625 g*L(-1)-25.00 g*L(-1), the inhibition rate was 9.28%-64.87%. In the sugar tolerance test, the blood sugar value in starch-loaded mice decreased sharply (P<0.01), followed by sucrose-loaded group (P<0.05), and there was no change in glucose-loaded group (P>0.05). In diabetic mice CEW-40 and CEW-10 groups showed significant blood sugar lowering effect (P<0.01 or P<0.05)., Conclusion: CEW has stronger effect in inhibition of AG activity than acarbose. CEW can increase the sugar tolerance in normal mice and decrease the blood sugar level in diabetic mice..

Objective: To evaluate the correlation of neutral alpha-glucosidase in seminal plasma with the location of epididymal obstruction in azoospermia men., Methods: We detected neutral alpha-glucosidase activity in the seminal plasma of 59 men with obstructive azoospermia followed by determining the location of epididymal obstruction by scrotal exploratory surgery. Then we analyzed the correlation between neutral alpha-glucosidase and the location of epididymal obstructive azoospermia., Results: Among the total number of patients, there were 25 cases of bilateral cauda epididymal obstruction, 15 bilateral corpus, 12 bilateral caput, 4 unilateral caput-opposite cauda, and 3 unilateral corpus-opposite cauda. The neutral alpha-glucosidase levels in the seminal plasma of bilateral cauda, corpus and capus epididymal obstructions were (4.1 +/- 1.9), (13.8 +/- 4.4) and (46.8 +/- 19.3) mU per ejaculate, respectively, with statistically significant differences among the three groups (P < 0.05)., Conclusion: Neutral alpha-glucosidase activity is significantly correlated with the location of epididymal obstruction in azoospermia men, which helps to locate epididymal obstruction, evaluate surgical prognosis and reduce the time of scrotal exploratory surgery.

The enzyme-inhibitor model and the sugar tolerance mouse model were used to evaluate the relationship between the inhibition rate of enzyme activity and concentration of Hippophae rhamnoides L. subsp. chinensis Rousi polysaccharide (HRP). The inhibitory patterns of enzyme and dose-dependent effects of HRP's effect on blood glucose using acarbose tablets as control were also examined. The mechanism underlying hypoglycemic effects of HRP was discussed. The results showed: in the enzyme-inhibitor model, the inhibitory activity of different concentrations of HRP (9.80, 19.60, 39.20, 78.40, 156.80 and 312.50 mg x L(-1)) on alpha-glucosaminidase (AG) inhibitory activity were 6.62%, 18.02%, 33.26%, 48.23%, 62.11%, 76.31%, 90.12%, IC50 was 31.59 mg x L(-1). The inhibitory rate of 25.00 x 10(3) mg x L(-1) acarbose tablets was only 64.87%, and IC50 was 10.75 x 10(3) mg x L(-1). In the sugar tolerance mouse model, different doses of HRP (240, 480, 960 mg x kg(-1)) tended to decrease levels of blood glucose compared with control group (acarbose tablets 375 mg x kg(-1)) at 15, 30, 60 and 120 min. It's further confirmed that HRP is a kind of competitive inhibitor of AG activity. Its inhibition rate increases with the increase of concentration in normal mice, and it subsequently improves the sugar tolerance showing the effect of reducing blood sugar.

Objective: To investigate the effects of icariin on the streptozocin (STZ)-induced epididymis impair in rats., Methods: The epididymis impair was induced by injection of streptozocin at dosage of 60 mg/kg ip in SD rats. Animals were randomly divided into six groups (n = 14): normal control, model group, three icariin treated group with different dosages (P.O, 20 mg/kg, 40 mg/kg, 80 mg/kg especially) and rosiglitazone group (P.O, 3 mg/kg), 12 weeks later, animals were sacrificed. The level of serum glucose, the activity of lactate dehydrogenase (LDH), acid phosphatase (ACP), gamma-glutamyltranspeptidase (gamma-GT), alpha-glucosidase activity as well as sialic acid (SA), fructose level in the epididymis were determined. The pathological examination was performed under microscope after the epididymis was fixed by 4% poly-formalin and stained by HE., Results: Compared with the normal control, the activity of LDH, ACP, gamma-GT and alpha-glucosidase in the epididymis revealed a decline, with lower level of SA and fructose. Histological examination showed that mature spermatocytes in the epididymis markedly decreased. These alterations were ameliorated in the groups with the treatment of icariin at 40 mg/kg and 80 mg/kg, but not at 20 mg/kg., Conclusion: Icariin ameliorated the signs of epididymis in diabetic rats induced by streptozocin, this effect might carry out by promoting the elevation of special enzyme and energy metabolism in the epididymis.

Diabetes is a global threat threatening human health in the world, with an increasing incidence rate in recent years. The disorder of glucose metabolism is one of the major factors. As relevant glucose metabolic enzymes such as alpha-glucosidase, glucose-6-phosphatase (G-6-P), glycogen phosphorylase (GP) and glycogen synthase kinase-3 (GSK-3) get involved in and control the process of glucose metabolism, the regulation of the activity of glucose metabolic enzymes is of significance to the treatment of diabetes. Traditional Chinese medicines (TCMs) have been widely researched because of their low toxicology and high efficiency, and many extracts and components from TCMs have been proven to be regulators of glucose metabolic enzymes. Compared with anti-diabetic western medicines, anti-diabetic TCMs feature safety, reliability and low price. This essay summarizes the anti-diabetic effect of TCMs on regulating glucose metabolic enzymes.

Objective: To investigate whether high-fat diet induced obesity was associated with variation of glucose absorption in small intestinal mucosa of rats., Methods: 46 male SD rats were randomly divided into high-fat diet group (n = 31) and control group (n = 15), fed with high-fat diet and normal diet for 24 weeks, respectively. After 24 weeks, the rats were divided into obese (n = 16) and obesity-resistant group (n = 10) according to their body weight. Rats' body weight, abdominal fat weight, plasma glucose level, maltase, sucrase activity in small intestinal mucosa were measured. SGLT-1 expression in intestinal mucosa was detected by immunohistochemistry, RT-PCR and Western blot., Results: Mean body weight, abdominal fat weight, fast plasma glucose levels, maltase activities and SGLT-1 protein expression in intestinal mucosa of obese rats were significantly higher than those in the control and obesity-resistant rats (P < 0.05). Sucrase activities in intestinal mucosa showed no statistical difference among the three groups (P > 0.05). The SGLT-1 mRNA expression in obese group was increased by 12.5% and 23% when compare with the control and obesity-resistant group, respectively. But the difference was not statistical significant (P > 0.05)., Conclusion: High-fat diet induced obesity was associated with the increased intestinal maltase activity and expression of SGLT-1 in rats, the key molecule in glucose absorption.

Objective: To increase the leavening ability in the lean dough, the maltose utilization ability of baker's yeast was enhanced., Methods: A 1.7kb PGK1 promoter and terminator were ligated and inserted into vector Yep-C to give the expression plasmid named Yep-CP. Then a 1.7kb DNA fragment containing the open reading frame and terminator of mal62 gene was amplified from Saccharomyces cerevisiae BY-14 by PCR, and inserted into Yep-CP to generate recombinant plasmid Yep-CPM. To express mal62 gene properly in S. cerevisiae, the recombinant expression plasmids Yep-CPM with copper resistance gene as the selection marker for yeast transformation were introduced into S. cerevisiae BY-14. The resulting yeast transformant BYCPM was screened on YEPD with 4 mmol/L CuSO,and identified by colony PCR. Target protein was detected by qRT-PCR, and the enzyme activities and the leavening ability of the recombinant strain BYCPM were determined to confirm whether functional expression was achieved., Results: The maximum maltase activity of recombinant strain BYCPM was 15%-52% higher than that of control strain BY-C. Leavening ability and specific leavening ability of recombinant strain BYCPM were 40% and 5.6% higher than that of the control strain BY-C, respectively., Conclusion: The mal62-overexpression was an effective way of increasing the maltase activity and the ability of glucose depression in the baker's yeast. Moreover, recombinant strain BYCPM could further enhance the leavening ability and produce more gas while consume lesser carbon source than the control strain.

The present study was conducted to develop new inhibitors of pancreatic lipase and alpha-glucosidase from Chinese dietary herbs. Sixty-three dietary herbs from 39 taxonomic families were selected and extracted with aqueous ethanol or water. The extracts were then tested with in vitro enzyme assays for their ability to inhibit pancreatic lipase and alpha-glucosidase activities. Orlistat and acarbose were used as two positive controls. The extracts of Nelumbo nucifera, Curcuma longa, Piper longum and Morus alba showed strong pancreatic lipase inhibitory effects with IC50 at (28.00 +/- 5.51), (5.24 +/- 0.51), (14.76 +/- 2.58), (4.78 +/- 0.58), (3.41 +/- 0.67) mg x L(-1), respectively. These extracts also showed potent alpha-glucosidase inhibitory activities with IC50 at (1.98 +/- 0.13), (0. 18 + 0.007), (0.71 +/- 0.08), (0.077 +/- 0.005), (0.089 +/- 0.006) g x L(-1), respectively. The results provide useful information for developing new drugs or natural health products for hyperlipidemia and hypoglycemia from Chinese dietary herbs.

Objective: To carry out prenatal diagnosis for a glycogen storage disease type II (GSD II ) affected family., Methods: The acid-α -glucosidase (GAA) activity was measured in whole leukocytes and cultured amniocytes with 4-methylumbelliferyl-α -D-glucopyranoside as substrate and with acarbose as inhibitor. The coding regions of GAA gene were amplified by polymerase chain reaction and analyzed by direct DNA sequencing., Results: The proband and the fetus had low GAA activity (12.3% and 1.1% of the average normal range, respectively). Mutation analysis of the GAA gene revealed a novel nonsense mutation p.W738X and a reported nonsense mutation p.E888X in both the proband and the fetus; the reported pseudodeficiency allele c.[1726G to A: 2065G to A] was found in the proband, the mother and the fetus., Conclusion: The proband and the fetus were both GSD II affected. A combination of GAA activity analysis and mutation analysis is efficient for the prenatal diagnosis of GSD II. Mutation analysis should be a routine method in the prenatal diagnosis of GSD II in Asian population, where pseudodeficiency allele can cause low GAA activity in normal individuals which is relatively common in Asian.

Twenty five new beta-aminoalcohols containing nabumetone moiety were prepared via the reduction of potassium borohydride with a convenient and efficient procedure, starting from beta-aminoketones that have been synthesized by our group. Their chemical structures were determined by IR, MS, 1H NMR, 13C NMR, HR-MS and antidiabetic activities were screened in vitro. Preliminary results revealed that the antidiabetic activity of most beta-aminoalcohols were better than that of the corresponding beta-aminoketones. Although most compounds showed weak antidiabetic activity, the alpha-glucosidase inhibitory activity of compounds 5hd(1) and 5id(2) reached 74.37% and 90.15%, respectively, which were superior to the positive control. The relative peroxisome proliferator-activated receptor response element (PPRE) activity of five compounds were more than 60%, among them compound 5ca possessed the highest activity (112.59%). As lead molecules of antidiabetic agents, compounds 5hd(1), 5id(2) and 5ca deserve further study.

Objective: To investigate the effect of epigallocatechin-3-gallate (EGCG) on the activity of alpha-glucosidase derived from bakers' yeast and small intestinal mucosa of Wistar rats, in order to establish an evaluation system for the activity of alpha-glucosidase inhibitor., Methods: alpha-glucosidase was prepared from the bakers' yeast and the small intestinal mucosa of Wistar rats. The inhibitory activity of EGCG on alpha-glucosidase was based on the concentration of glucose in the reaction system., Results: EGCG was shown to be an alpha-glucosidase inhibitor. IC50 was 0.150 mg/ml and 0.837 mg/ml for alpha-glucosidase derived from bakers' yeast and small intestinal mucosa respectively., Conclusion: The establishment of an in vitro evaluation system for AGI activity is conductive for screening alpha-glucosidase inhibitory substance. The AGI activity of EGCG is strong in vitro.

Searching for new antidiabetic lead compound, 4-(1-aryl-3-oxo-5-phenylpentylamino) benzenesulfonamides were designed and synthesized directly by three component one-pot condensation of 4-phenyl-2-butanone and sulfanilamide with some aromatic aldehydes at an yield of 23%-97%. The chemical structures of the twelve new Mannich bases were confirmed by 1H NMR, 13C NMR, FTIR, ESI-MS and HR-MS. The screening results of antidiabetic activity indicated that most of these title compounds possess alpha-glucosidase inhibitory activity, among which compound le is the strongest one. And compound 11 possesses good peroxisome proliferator-activated receptor response element (PPRE) agonist activity. The structure-activity relationship of these new beta-amino ketones containing benzenesulfonamide unit was also discussed preliminarily.

Objective: To study the changes of neutral alpha-glucoside activity in the epididymis of heroin-dependent and heroin-withdrawal rats, and to investigate the effects of intervention with purine nucleotide (AMP and GMP)., Methods: Eighty Wistar rats were randomly divided into 8 groups of equal number, control, nucleotide, heroin, heroin + nucleotide, 3 d withdrawal, 9 d withdrawal, 3 d nucleotide (nucleotide administrated for 3 days after heroin withdrawal) and 9 d nucleotide (nucleotide administrated for 9 days after heroin withdrawal). Neutral alpha-glucosidase activity in the epididymis was detected in each group of rats., Results: Compared with the control group, neutral alpha-glucoside activity was markedly decreased in the heroin group (P < 0.05), and also in the 3 d and 9 d withdrawal groups, although with no significant differences (P > 0.05)., Conclusion: Heroin reduces neutral alpha-glucoside activity in the epididymis of rats, and this effect may continue for some time after drug withdrawal, while purine nucleotide can keep neutral alpha-glucosidase activity in a relatively stable state.

Objective: To search alpha-glucosidase inhibitors from Rubia cordifolia., Method: The alpha-glucosidase inhibitors were isolated by the column chromatographic techniques and the bioassay-guided method in vitro. A combination of MS and NMR spectroscopy was used to identify the chemical structures. The inhibitory kinetics of the inhibitors were also investigated., Result: The chloroform extract showed high inhibitory activity, and three active compounds were isolated and identified as 1,3-dihydroxy-2-methylanthraquinone (1), 1-hydroxy-2-methylanthraquinone (2) and 1,2-dihydroxyanthraquinone (3). The IC5o values of compound 1-3 were all lower than that of acarbose. Compound 1 and 2 shown competitive type manner on alpha-glucosidase, whereas compound 3 shown noncompetitive type model., Conclusion: Compounds 1-3 as strong inhibitors of alpha-glucosidase were reported for the first time.

Objective: Alpha-glucosidase from Aspergillus niger SG136 was expressed in Pichia pastoris., Methods: cDNA of mature alpha-glucosidase (aglu) was amplified from the total DNA of A. niger SG136 by PCR and overlap-PCR with primers designed based on the sequence of A. niger CBS 513.88 in NCBI database. The gene was cloned into pMD18-T simple vector. The sequencing result showed that the gene encoded for a protein of 960 amino acids residues, which was 1 amino acid different from that of A. niger CBS 513.88. The expression vector pPIC9K-aglu was constructed by subcloning the gene into plasmid pPIC9K, and then transformed into P. pastoris through electroporation after linearized by Bgl II digestion. The recombinant P. pastoris KM71/pPIC9K-aglu were screened in MD and YPD/G418 plates,and identified by PCR. In shaking culture condition, methanol was added to a final concentration of 1% to induce the secretion of alpha-glucosidase., Results: Electrophoresis analysis of KM71/pPIC9K-aglu culture supernatant showed that there were two major protein bands corresponding to 98 kDa and 33 kDa respectively in SDS-PAGE, and there was only one band in Native PAGE; while in the control experiment of KM71/pPIC9K, there were no visible bands. Transglucosidation reaction from crude enzyme revealed that contents of isomaltooligosaccharides were up to 26.0% under the optimal conditions of pH 5 and 60 degrees C at 24 h., Conclusion: A. niger alpha-glucosidase was expressed in P. pastoris with transglucosidation activity.

Objective: To search alpha-glucosidase inhibitors from Luculia pinciana., Method: The alpha-glucosidase inhibitor was isolated by the column chromatographic techniques and the bioassay-guided method in vitro. A combination of MS and NMR spectroscopy was used to identify the chemical structures. The inhibitory kinetic of the isolations was also investigated., Result: The ethyl acetate extract showed strongest inhibitory activity, and five active compounds were isolated and identified as scopletin (1), 5-methoxy-8-hydroxycoumarin (2), lalpha,3beta,24-trihydroxyursolic acid (3), ursolic acid (4) and oleanolic acid (5). The IC50 values of all compounds were lower than that of acarbose. Four of them shown noncompetitive type manner on alpha-glucosidase inhibition except that compound 3 is competitive type manner., Conclusion: Compounds 1-4 as the inhibitors of alpha-glucosidase were reported for the first time.

In order to find highly active antidiabetic lead compound, sixteen 4-aminobenzoic acid derivatives were designed and synthesized directly through Mannich reaction in the solution of ethanol at 15-35 degrees C with facile method, mild reaction condition and high yield (45%-90%). Fifteen of them are new compounds. Their structures were confirmed by 1H NMR, 13C NMR, IR, ESI-MS and HR-MS. Alpha-glucosidase inhibitory activity of these compounds indicated that most of these compounds possess the activity with the order: 2c > 2b > 2h > 1a > 1f. The structure-activity relationship of these 4-aminobenzoic acid derivatives was also discussed.

Objective: To study the inhibitive effect of extracts from Scindapsus aureus on alpha-glucosidase and its antioxidant activity., Methods: The 70% ethanol extracts was extracted by petroleum, ethyl acetate and n-butanol, and the inhibitory activity against alpha-glucosidases and antioxidative effects of each fraction were determined in vitro, and the inhibitory kinetics of ethyl acetate fraction was investigated., Results: The inhibitory activity of ethly acetate fraction against alpha-glucosidase was higher as its IC50 was 43.63 microg/mL. The inhibition kinetics analyzed by Hnewaver-Burk plots showed that ethyl acetate fraction was a competitive type inhibitor, and the inhibition constants for free ennzyme (Ki) was determined to be 21.49 microg/mL. The antioxidative effects of ethly acetate and n-butanol fractions exhibited stronger than that of other fractions, the scavenging ability on DPPH at the concentration of 0.42 mg/mL were 88.5% and 93.6%. Ethly acetate fraction showed scavenging ability on autioxidantor, and the IC50 values was 0.27 mg/mL., Conclusion: Ethyld acetate fraction of extract from Scindapsus aureus has potential inhibitroy activity against alpha-glucosidase and antioxidant effects, and n-butanol fraction has great antioxidative effects.

Objective: To compare alpha-glycosidase inhibitor microamount screening models which are established by two different resources alpha-glycosidase to screen the Chinese herbal medicines which have great alpha-glycosidase inhibition., Methods: Comparing the activities of two different resources alpha-glycosidase with glucose oxidase method, then establishing optimal reaction conditions. Extracting the water soluble compositions of Chinese gallnut, paeoniae radix, Glycyrrhiza, rhubarb, fructus by boiling ,then using the alpha-glycosidase inhibition model to check and compare their alpha-glycosidase inhibitions., Results: Five Chinese herbal medicines all had alpha-glycosidase inhibition, and the greatest was Chinese gallnut., Conclusion: The activities of two resources alpha-glycosidase are certainly different, but they do not have significant influence on findings of screening alpha-glycosidase inhibitors in vitro. Water soluble compositions of five Chinese herbal medicines have alpha-glycosidase inhibition.

Objective: To understand Ureaplasma urealyticum (Uu) infection, analyzed the influence of Uu infection on the seminal quality and the accessory genetical gland function in male infertility patients, and investigate its mechanism., Methods: We cultured 202 semen samples collected from male infertility patients and analyzed the influence of Uu infection on seminal parameters and the biochemical indexes of the seminal plasma., Results: The Uu infection rate was 33.7% in the infertile males, with no statistic differences between the Uu positive and negative groups either in the average age (28.9 +/- 4.7 yrs vs 29.6 +/- 4.0 yrs, P = 0.250) or in the seminal quantity (2.93 +/- 1.32 ml vs 2.86 +/- 1.52 ml, P = 0.774). The sperm density, motility and vitality were (84.37 +/- 52.92) x 10(6) ml, (44.62 +/-22.13) % and (38.40 +/- 15.61) % in the Uu positive group, significantly lower than (101.90 +/- 43.90) x 10(6) ml, (51.83 +/- 19.88) % and (44.45 +/- 15.47) % in the Uu negative group (P = 0.025, P = 0.036 and P = 0.020). The seminal pH value was normal in both of the groups, but significantly higher in the Uu positive than in the negative group (7.32 +/- 0.10 vs 7.19 +/- 0.29, P = 0.003). VCL, VSL, VAP and MAD were significantly lower, while BCF was significant higher in the former than in the latter [(33.97 +/- 8.96) microm/s vs (39.70 +/- 8.14) microm/s, t = 4.113, P < 0.001; (22.29 +/- 6.06) microm/s vs (25.20 +/- 6.67) microm/s, t = 2.684, P = 0.008; (25.96 +/- 6.83) microm/s vs (30.02 +/- 6.81) microm/s, t = 3.537, P < 0.001; 46.60 +/- 13.68 vs 54.23 +/- 15.14, t = 3.112, P = 0.002; (6.12 +/- 1.89) Hz vs (5.22 +/- 1.64) Hz, t = 3. 164, P = 0.002]. All the five indexes were influenced by Uu infection. Compared with the negative group, the seminal plasma alpha-glucosidase was significantly decreased in the positive group [(40.0 +/-18.7) U/ml vs (47.9 +/- 21.0) U/ml, t = 2.248, P = 0.026], and the risk of the decrease was 2.12 times higher. No statistic difference was observed in seminal plasma acid phosphatase and seminal plasma fructose between the two groups., Conclusion: Uu infection in the genital tract is an important factor of seminal quality reduction in infertile men and may cause a decreased secretion of alpha-glucosidase in the epididymis, but it hardly influences the prostate and seminal vesicle.

Objective: To establish a method of determining alpha-glucosidase activity in seminal plasma by semi-automatic biochemistry analyzer., Methods: Alpha-glucosidase activity in seminal plasma from 51 men with normal semen parameters in routine semen analysis were detected by semi-automatic biochemistry analyzer and manual glucose oxidase method, respectively. Then, the intra-assay and inter-assay coefficient variation (CV) and normal reference value were calculated. In the meanwhile, the correlation between the two methods was analyzed., Results: The intra-assay CVs of 2 seminal plasma samples with different alpha-glucosidase activity detected by semi-automatic biochemistry analyzer were 12.63% and 9.13%, and the inter-assay CVs were 10.67% and 13.49%, respectively. The normal reference value for seminal alpha-glucosidase activity detected with semi-automatic biochemistry analyzer ranged from 102.28 to 555.08 U/L. There was a significantly positive correlation between the semi-automatic biochemistry analyzer and the manual glucose oxidase method (r = 0.792, P < 0.01)., Conclusion: The method of determining alpha-glucosidase activity in seminal plasma by semi-automatic biochemistry analyzer, with its simplicity, less cost of time and reagents, and more reliable result, could be applied to clinical laboratory medicine.

Objective: To study the hypoglycemic effect of the extract of B. polyandra (SHG)., Method: The diabetic mice were induced by alloxan in ICR mice. The blood glucose concentration was measured by glucose oxidase method. The serum insulin level was determined by 125I-insulin radioimmunoassay kit. The hypoglycemic effect was evaluated by the levels of both fasting and no-fasting blood glucose. The effect on serum insulin level was estimated by the values of the blood insulin and the changes of the blood glucose induced by the glucose intraperitoneal injection. The effect on the glucose absorption was investigated by the oral sucrose or starch tolerance test., Result: Both of the fasting and no-fasting blood glucose levels were decreased significantly by the treatment of 20 or 30 g raw materials crude drug x kg (-1) SHG orally for 7-10 d in ICR mice or in alloxan diabetic mice. In the oral sucrose tolerance test or oral starch tolerance test, the administration of SHG reduced significantly the peak value of the blood glucose and the area under the blood glucose-time curve (AUC) in normal or alloxan diabetic mice, respectively. These effects of SHG were similar to those of acarbose, a kind of alpha-glucosidase inhibitors. In the oral glucose tolerance test in normal and alloxan diabetic mice, SHG decreased both the blood glucose peak and the AUC induced by the glucose loading. But in the intraperitoneal injection glucose tolerance test the levels of insulin in both SHG and control mice were similar, however, the changes of the blood glucose level after the glucose-loading for 30 min in SHG mice was much lower than that in control mice., Conclusion: With the treatment of SHG, the fasting and no-fasting blood glucose concentrations were decreased and the glucose tolerance improved significantly in both normal and alloxan diabetic mice, and the inhibition of a-glucosidase might be one of its major mechanisms.

Objective: To investigate the inhibition effect of Psidium guajava linn (PGL), a leaf water-soluble extract, on the activities of alpha-glucosidases., Methods: The PGL water-soluble extract (PGL WE) was obtained by the procedure of distilled water immersion, filtration, extracted fluid concentration and dry of Psidium guajava leaf. The diabetes of Kunming mice was induced by intraperitoneal injection of Streptozotocin (STZ). The small intestinal mucosa of diabetic mice was scraped to make the homogenate for the preparation of alpha-glucosidases. In vitro, the homogenates were incubated with sucrose and maltose. The formed glucose represented the activities of alpha-glucosidases. The Lineweaver-Burk plot was applied to determine the type of alpha-glucosidase activity inhibited., Results: The water-soluble extract from PGL significantly inhibited, in the dose-dependent manner, the activities of alpha-glucosidase from small intestinal mucosa of diabetic mice. The PGL extract inhibition concentration (IC50) to sucrase or maltase was 1.0 g/L or 3.0 g/L respectively. The mixed inhibition type was showed to be the competitive and non-competitive inhibition., Conclusion: The GPL water-soluble extract possesses the potential effect of inhibition on the alpha-glucosidase activity from the small intestinal mucosa of diabetic mouse.

Objective: To study the relationships of experimental varicocele to the apoptosis of epididymis epithelium and changes of the contents of alpha-1,4-glucosidase and sialic acid from the unilateral epididymis in adolescent rats, and to investigate the effects of varicocele on the unilateral epididymis epithelium., Methods: Experimental left varicocele models of 16 adult male Sprague-Dawley rats were obtained by partial ligation of the left renal vein. The epididymides were collected for detecting the apoptosis of epididymis epithelium and the contents of alpha-1,4-glucosidase and sialic acid by using spectrophotometry., Results: Seven days after the establishment of the left varicocele model, the index of the apoptosis of the left epididymis epithelium was significantly higher (P < 0.001) and the contents of alpha-1,4-glucosidase and sialic acid significantly lower (P < 0.05, P < 0.005) in the experimental group than in the control., Conclusion: The results suggest that unilateral varicocele may increase the apoptosis of epididymis epithelium and the contents of alpha-1,4-glucosidase and sialic acid and subsequently affect the synthesizing and secretory function of the epididymis.

Objective: To determine the effects of saponins from Tribulus terrestris (STT) on small intestinal a-glucosidase and postprandial blood glucose levels in rats., Method: The inhibitory effects of STT on a-glucosidase extracted from small intestines in rats were carried out in vitro. The blood glucose levels were measured after 60 min when sucrose (2 g x kg(-1)) or glucose (2 g x kg(-1)) was administered orally with STT (100 mg x kg(-1)). After treated with STT (100 mg x kg(-1)) for 14 d, the activities of a-glucosidase were determined daily, as well as the postprandial blood glucose levels after oraly administered sucrose (2 g x kg(-1))., Result: STT at concentrations of 0.1, 1 and 10 mg x mL(-1) reduced significantly the activities of alpha-glucosidase with inhibitory rates of (20.83 +/- 1.66)%, (43.73 +/- 2.39)% and (52.62 +/- 2.69)%, respectively. In facts STT (100 mg x kg(-1)) considerably decreased the blood glucose levels which was 52.61% of that of the control in rats co-administered orally with sucrose (2 g x kg(-1)). However, it showed no such effect on the rats co-administered orally with glucose (2 g x kg(-1)). After orally administered of STT for 14 d, the activity of alpha-glucosidase was significantly reduced (P < 0.05) to (58.17 +/- 3.24)% of that those in control. Meanwhile, The rats were oral administered with sucrose, the increase of postprandial blood glucose levels were (69.50 +/- 4.28)% of that in control 60 min later ( P < 0.05)., Conclusion: It was through inhibiting the activity of a-glucosidase in small intestines that STT significantly retarded the increase in postprandial blood glucose levels in rats.

Objective: To search for glucosidase inhibitors of various fractions extracted from mulberry leaves., Method: The constituents of mulberry leaves water fraction 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. To investigate in vivo effect of lowering blood glucose, the mouse blood glucose level was assayed by glucose tolerance experiments., Result: The glucosidase inhibitory activities were found in all the constituents of alkaloids, flavones and amyloses, the alkaloid constituent being the strongest., Conclusion: The effect of reducing blood glucose of mulberry leaves is related to the inhibitory activities against glucosidase of different constituents.

Objective: To investigate the levels and changes of sexual hormone in serum and seminal plasma, and the epidermal growth factor (EGF), alpha-glucosidase, and fructose in the seminal plasma of non-obstructive azoospermic patients, and to assess the significance of the analysis of their chromosomes., Methods: Thirty-six non-obstructive azoospermic males and 26 normal males were chosen. The results in reproductive endocrine and genetics obtained by radioimmunoassay, colorimetry, and chromosomes G and C banding assay were compared and analyzed to explore into the causes of infertility., Results: The levels of FSH, LH, PRL and E2 in the serum of the non-obstructive azoospermic patients were higher than those of the control group, but the T level had no significant difference between the two groups. There were no obvious differences in the sexual hormones except in FSH of the seminal plasma. There were 2 cases of abnormal sex chromosomes, 2 cases of big Y chromosomes and 7 cases of small Y chromosomes in the non-obstructive azoospermic patients. No correlation was found between EGF and fructose in the seminal plasma., Conclusion: The level of sexual hormone in serum, alpha-glucosidase in seminal plasma and abnormal chromosomes might be closely related to non-obstructive azoospermia.