小麦淀粉生物制糖工艺--发明专利工艺技术介绍

介绍了北京坡华公司一种小麦淀粉生物制糖的发明专利技术,在小麦淀粉生产葡萄糖液用于生物发酵营养基质的工艺过程中,此技术可提高小麦淀粉的利用率,提高相关企业的经济效益。

Purification and Characteristics of Mn-containing Nitrogenase Component Ⅰ

A mutant UW 3, which is unable to fix N 2 in the presence of Mo (Nif -) but undergo phenotypic reversal to Nif + under Mo deficiency, was able to grow in Mo- and NH 3-deficient medium containing Mn, and the growth was accelerated by Mn at low concentration. A partly purified nitrogenase component Ⅰ protein separated from UW 3 grown in the Mn-containing medium was shown to contain Fe and Mn atoms (ratio of Fe/Mo/Mn: 10.41/0.19/1.00) with C 2H 2- and H +-reducing activity which almost equal to half of that of MoFe protein purified from wild-type mutant of Azotobacter vinelandii Lipmann. This protein was obviously different from MoFe protein in both absorption spectrum and circular dichroism, and the molecular weight of subunits in Mn-containing protein was close to that of α subunit in MoFe protein. The preliminary results indicated that the protein containing Mn might be a nitrogenase component Ⅰ protein.

Synthesis of Chiral Polyaniline Induced by Modified Hemoglobin

The synthesis of chiral polyaniline （PANI） induced by modified hemoglobin （Hb） was pro- foundly explored for the first time. Results revealed that after being separated, inactivated or immobilized, Hb can still induce the formation of chiral PANI successfully, suggesting that Hb can be used as the chiral inducers regardless of harsh reaction conditions. By examining the properties of PANI induced by modified Hb, it was found that Hb（inactivated）-PANI possessed excellent chirality, stability, and crystalline structure. The globin separated from Hb was demonstrated to have the ability of inducing the production of chiral PANI whereas the hematin from Hb had no capacity to direct enantio specificity for the PANI chains. Results indicated that Hb（immobilized）-PANI exhibited poor yield, doping state, and crys- talline structure, indicating that the immobilization of Hb by entrapment was not beneficial to the polymerization reaction. Results also showed that the structure of Hb may have significant effects on the morphologies of chiral PANI.

Purification and Characterization of Cr-containing Nitrogenase Component Ⅰ

A mutant UW 3, which is unable to fix N 2 in the presence of Mo (Nif -) but can undergo phenotypic reversal to Nif + under Mo_deficient conditions, was able to grow in Cr_containing but Mo_ and NH 3_deficient medium. A partly purified nitrogenase component Ⅰ protein obtained from UW 3 grown on the Cr_containing medium was shown to contain Fe and Cr (atom ratio of Fe to Cr and Mo to Cr: 11.60 and 0.41) and to have 70% of the C 2H 2_ and H +_reduction activity of MoFe protein from the wild_type strain of Azotobacter vinelandii Lipmann. The Cr_containing protein was different in subunit composition from that of MnFe protein purified from the mutant strain grown in the presence of Mn, but similar to that of MoFe protein, that is, it was a tetramer composed of two different subunits (α 2β 2). The preliminary results indicated that the Cr_containing protein might be a nitrogenase component Ⅰ protein.

Attainment of multifactorial treatment targets among the elderly in a lipid clinic

Objective To examine target attainment of lipid-lowering, antihypenensive and antidiabetic treatment in the elderly in a specialist set- ting of a University Hospital in Greece. Methods This was a retrospective study including consecutive subjects 〉 65 years old （n = 465） with a follow-up 〉 3 years. Low-density lipoprotein cholesterol （LDL-C）, blood pressure （BP） and glycated hemoglobin （HbAlc） goal achievement were recorded according to European Society of Cardiology/European Atherosclerosis Society （ESC/EAS）, European Society of Hypertension （ESH）/ESC and European Association for the Study of Diabetes （EASD） guidelines. Results The LDL-C targets were attained by 27~,4, 48% and 62% of very high, high and moderate risk patients, respectively. Those receiving statin ＋ ezetimibe achieved higher rates of LDL-C goal achievement compared with those receiving statin monotherapy （48% vs. 33%, P 〈 0.05）. Of the diabetic sub- jects, 71% had BP 〈 140/85 mmHg, while 78% of those without diabetes had BP 〈 140/90 mmHg. A higher proportion of the non-diabetic individuals （86%） had BP 〈 150/90 mmHg. Also, a higher proportion of those with diabetes had HbAlc 〈 8% rather than 〈 7% （88% and 47%, respectively）. Of note, almost one out of three non-diabetic individuals and one out of ten diabetic individuals had achieved all three treatment targets. Conclusions Even in a specialist setting of a University Hospital, a high proportion of the elderly remain at suboptimal LDL-C, BP and HbAlc levels. The use of drug combinations could improve multifactorial treatment target attainment, while less strict tar- gets could be more easily achieved in this population.

Purification and Activation In Vitro of MoFe Protein from a nifE Deleted Mutant Strain of Azotobacter vinelandii

The Deltanif E MoFe protein (Deltanif E Av1) was obtained by a chromatography on DE52, Sephacryl S-300 and Q-Sepharose columns from the unheated crude extract of nifE-deleted mutant strain (DJ35) of Azotobacter vinelandii Lipmann. The analysis by SDS-PAGE showed that the Delta nif EAv1 was similar to OP MoFe protein (Av1) of A. vinelandii in the kinds and molecular weights of subunits (alpha and beta subunit). When complemented with nitrogenase Fe protein (M), the A nif EAv1 had hardly any proton-reduction activity, but could be significantly activated by FeMoco extracted from OP Av1. After the Delta nif E Av1 was treated with an excess o-phenanthroline (o-phen) and chromatographied on Sephadex G-25 column under atmosphere of Ar, Delta nif E Av1(C) was obtained. In the presence of both Av2 and MgATP regeneration system, the Delta nif EAv1(C), rather than A nif EAv1, was significantly activated in vitro by a reconstituent solution containing Mn which composed of KMnO4, ferric homocitrate, Na2S, Na2S2O4 (DT) and dithiothreitol (DTT). But in the absence of MgATP or Av2, the activation of Delta nifE Av1(C) did not happen. It indicates that activation of Delta nif EAv1 by RS-Mn requires the pretreatment with o-phen and the simultaneous presence of Av2 and MgATP.

Number of medications is associated with outcomes in the elderly patient with metabolic syndrome

Background The diagnosis of metabolic syndrome indicates a clustering of metabolic imbalances which in sum have been recognized as a major predictor of cardiovascular and all-cause mortality. The aim of this study was to assess the level of under-pharmacy and poly-pbarmacy and its prognostic impact in elderly patients with metabolic syndrome. Methods Retrospective chart-review at a tertiary medical center, of 324 patients greater than 65 years of age who met the International Diabetes Foundation criteria for metabolic syndrome diagnosis [Body Mass Index （BMI） 〉 30 kg/m2, diagnosis of type 2 diabetes, hypertension, and dyslipidemia]. Results There were 60 （18.5%） patients in the low （〈 5） medication burden group, 159 （49.1%） in the medium （〉 5 and 〈 10） medication burden group, and 105 （32.4%） in the high （〉 10） medication burden group. At baseline, the groups differed only by systolic blood pressure. At two years follow-up, the medium group had significantly better improvement in high density lipoprotein cholesterol （HDL-C）, low density lipoprotein cholesterol （LDL-C）, HbAlc, and systolic blood pressure compared to the low medication burden group and significantly better improvement in triglycerides, Haemoglobin Alc （HbAlc） and systolic blood pressure compared to the high medication group. Decrease in HDL-C was the only variable associated with strokes. High medication burden predicted hospitalization burden. The number of anti-hypertensives, history of tobacco use, low and high medication burdens and decrease in HDL-C were all associated with death. Conclusions Both poly-pharmacy and under-pharmacy are associated with a decreased therapeutic benefit among patients with metabolic syndrome in terms of important laboratory measurements as well as clinical outcomes such as myocardial infarctions, hospitalization, and death.

Yi-Qi-Zeng-Min-Tang,a Chinese medicine,ameliorates insulin resistance in type 2 diabetic rats

AIM:To investigate the effects of the Chinese herbal decoction,Yi-Qi-Zeng-Min-Tang(YQZMT),on insulin resistance in type 2 diabetic rats.METHODS:Sprague-Dawley rats were divided into two dietary regiments by feeding either normal pellet diet(NPD) or high fat diet(HFD).Four weeks later,the HFD-fed rats were injected intraperitoneally with lowdose streptozotocin(STZ).Rats with non-fasting blood glucose level ≥ 16.67 mmol/L were considered type 2 diabetic and further divided into five subgroups:the type 2 diabetes model group,low-dose,medium-doseand high-dose YQZMT groups,and rosiglitazone group.Age-matched NPD-fed rats served as controls.YQZMT or rosiglitazone were administered for 8 wk.Intraperitoneal glucose and insulin tolerance tests were performed before and after the treatment to measure the glucose tolerance and insulin sensitivity.Serum levels of biochemical parameters,adipocytokines,such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),as well as free fatty acids(FFAs),were also analyzed.RESULTS:There was significant elevation of insulin resistance and serum levels of fasting glucose(12.82 ± 1.08 mmol/L vs 3.60 ± 0.31 mmol/L,P < 0.01),insulin(7197.36 ± 253.89 pg/mL vs 4820.49 ± 326.89 pg/mL,P < 0.01),total cholesterol(TC)(8.40 ± 0.49 mmol/L vs 2.14 ± 0.06 mmol/L,P < 0.01),triglyceride(2.24 ± 0.12 mmol/L vs 0.78 ± 0.05 mmol/L,P < 0.01),low-density lipoprotein cholesterol(LDL-c)(7.84 ± 0.51 mmol/L vs 0.72 ± 0.04 mmol/L,P < 0.01) and decrease in high-density lipoprotein cholesterol(HDL-c)(0.57 ± 0.03 mmol/L vs 1.27 ± 0.03 mmol/L,P < 0.01) in the low-dose STZ and high-fat diet induced type 2 diabetic group when compared with the control group.Administration of YQZMT induced dose-and timedependent changes in insulin resistance,glucose and lipid profile,and reduced levels of FFA,TNF-α and IL-6 in the type 2 diabetic rats.After the treatment,compared with the diabetic group,the insulin resistance was ameliorated in the high-dose YQZMT(2.82 g/100 g per day) group,with a significant reduction in serum glucose(12.16 ± 1.00 mmol/L vs 17.65 ± 2.22 mmol/L,P < 0.01),homeostasis model assessment of basal insulin resistance(22.68 ± 2.37 vs 38.79 ± 9.02,P < 0.05),triglyceride(0.87 ± 0.15 mmol/L vs 1.99 ± 0.26 mmol/L,P < 0.01),TC(3.31 ± 0.52 mmol/L vs 6.50 ± 1.04 mmol/L,P < 0.01) and LDL-c(2.47 ± 0.50 mmol/L vs 6.00 ± 1.07 mmol/L,P < 0.01),and a signif icant increase in HDL-c(0.84 ± 0.08 mmol/L vs 0.50 ± 0.03 mmol/L,P < 0.01).But the body weight was not changed signif icantly.CONCLUSION:YQZMT,which ameliorates insulin resistance and does not cause increase in body weight,may be a suitable therapeutic adjunct for the treatment of type 2 diabetes.

Effects of purified herbal extract of Salvia miltiorrhiza on lipid profile in hyperlipidemic patients

Background and Objective Hyperlipidemia is one of the most potent and best substantiated risk factors for coronary heart disease （CHD）. Purified Salvia miltiorrhiza extract （PSME） had been shown to have hypolipidemic effect in animal experiments. This study aimed to evaluate its lipids modulating effects in hyperlipidemic patients. Methods In this single-blind, placebo cantrolled study, lipid profiles of 80 hyperlipidemic patients were checked at same conditions. They were divided into two equal groups randomly （each composing of 40 patients）. They were given PSME tablet （800 mg） three times per day, or placebo tablet. All patients were put on NCEP type II diet and six weeks later, lipid profiles were checked. Results In PSME group, total cholesterol decreased by 27.32 mg/dl （12.3% reduction）, LDL-cholesterol decreased by 23.13 mg/dl （16.8% reduction） and HDL-cholesterol increased by 9.06 mg/dl （11.1%）, all were statistically significant. Although triglyceride dropped by 12.12 mg/dl （5.1%） but this was not significant statistically （P=0.34）. There were no significant changes oflipids levels in the placebo group. Conclusions PSME has significant favorable effect on total cholesterol, LDL-cholesterol, and HDL-cholesterol and may be a potential agent for the treatment ofatherogenic dyslipidemia.

A novel organic-inorganic hybrid monolith for trypsin immobilization

In proteomics, attention has focused on various immobilized enzyme reactors (IMERs) for the realization of high throughput digestion. In this report, a novel organic-inorganic hybrid monolith based IMER was prepared in a 100 μm i.d. capillary with 3-glycidoxypropyltrimethoxysilane (GLYMO) as the monomer and tetraethoxysilane (TEOS) as the crosslinker. Trypsin immobilization was achieved via the reaction between vicinal diol groups, which were obtained from hydrolysis of epoxy groups, and the amino groups of trypsin. Bovine serum albumin was digested thoroughly by this IMER in 47 s. After micro-reverse phase liquid chromatography-tandem mass spectrometry (μRPLC-MS/MS) analysis and database searching, beyond 35% sequence coverage was obtained, and the result was comparable to that of 12 h in solution digestion. The present IMER has potential for high throughput digestion.

An ultra-fast and highly efficient multiple proteases digestion strategy using graphene-oxide-based immobilized protease reagents

Highly efficient and rapid proteolytic digestion of proteins into peptides is a crucial step in shotgun-based proteome-analysis strategy. Tandem digestion by two or more proteases is demonstrated to be helpful for increasing digestion efficiency and de- creasing missed cleavages, which results in more peptides that are compatible with mass-spectrometry analysis. Compared to conventional solution digestion, immobilized protease digestion has the obvious advantages of short digestion time, no self-proteolysis, and reusability. We proposed a multiple-immobilized proteases-digestion strategy that combines the ad- vantages of the two digestion strategies mentioned above. Graphene-oxide （GO）-based immobilized trypsin and endoprotein- ase Glu-C were prepared by covalently attaching them onto the GO surface. The prepared GO-trypsin and GO-Glu-C were successfully applied in standard protein digestion and multiple immobilized proteases digestion of total proteins of Thermoan- aerobacter tengcongensis. Compared to 12-hour solution digestion using trypsin or Glu-C, 14% and 7% improvement were obtained, respectively, in the sequence coverage of BSA by one-minute digestion using GO-trypsin and GO-GIu-C. Multiple immobilized-proteases digestion of the total proteins of Thermoanaerobacter tengcongensis showed 24.3% and 48.7% en- hancement in the numbers of identified proteins than was obtained using GO-trypsin or GO-Glu-C alone. The ultra-fast and highly efficient digestion can be contributed to the high loading capacity of protease on GO, which leads to fewer missed cleavages and more complete digestion. As a result, improved protein identification and sequence coverage can be expected.

Polydopamine-mediated immobilization of phenylboronic acid on magnetic microspheres for selective enrichment of glycoproteins and glycopeptides

In this work, we demonstrate for the first time, a method to synthesize phenylboronic acid-Fe304@polydopamine （Fe3O4@ PDA-PBA） magnetic microspheres via the combination of mussel-inspired polydopamine coating and click chemistry. Uniform-size and core-shell structured Fe3O4@PDA-PBA magnetic microspheres with a core diameter of -240 nm and a shell thickness of -13 nm were obtained as identified by the characterization of the morphology, structure and composition of the synthesized microspheres. We evaluated the selectivity and binding capacity of the Fe3O4@PDA-PBA magnetic microsphcres by using standard glycoproteins （ovalbumin, immunoglobulin G and catalase） and nonglycoproteins （human serum albumin, bovine hemoglobin, myoglobin, lysozyme, and ribonuclease A） as model proteins. Adsorption experiments, SDS-PAGE and mass spectrometry analysis demonstrated that the Fe3O4@PDA-PBA magnetic microspheres had much high binding capacity and selectivity for glycoproteins/glycopeptides compared to nonglycoproteins/nonglycopeptides. In addition, the practicability of the Fe3O4@PDA-PBA magnetic microspheres was further assessed by selective capture of glycoproteins from healthy hu- man serum. The good results demonstrated its potential in glycoproteome analysis.

Solid-state electrochemiluminescence protein biosensor with aptamer substitution strategy

One solid-state electrochemiluminescence （ECL） protein biosensor based on the competing reaction and substitute reaction between protein-to-DNA aptamer and DNA-to-DNA aptamer was proposed. Additionally, the biosensor was based on ECL photo-quenching effect of ferrocene （Fc） to tris（2,2-bipyridyl）ruthenium（II） （Ru（bpy）2＋）. It was built up by modification of Au nanoparticles （AuNPs） and Ru（bpy）32＋ on one Au electrode firstly, and then self-assembly of one special double-stranded DNA （dsDNA） onto the electrode. This dsDNA was prepared by hybridization of one Fc labeled molecular beacon single-stranded DNA（ssDNA） and one anti-thrombin aptamer ssDNA. Without the target protein, this Fc-dsDNA/Ru（bpy）2＋- AuNPs/Au elec- trode trigged strong ECL signal, so we called it ECL ＂signal on＂ state. When thrombin was present in the sensing solution, the protein reacted with its aptamer from the Fc-dsDNA/Ru（bpy）3^2＋-AuNPs/Au electrode. Then the left molecular beacon ssDNA on the electrode recovered to its normal stem-loop structure and consequently its Fc labeler was close enough to the electrode surface to quench the ECL signal from Ru（bpy）3^2＋. It was in ECL ＂signal off＂ state. We measured the decrease in ECL intensity to sense the target protein. This was one endeavour to sense protein by using un-labeling target or probe strategy, which gave higher sensitivity and selectivity due to the better combination efficiency of protein and the un-labeled aptamer. 6.25 fmo/L thrombin was detected out,

Antioxidant and hepatoprotective effect of different extracts of Guizhencao (Herba Bidentis Bipinnatae) against liver injury in hyperlipidemia rats

OBJEVTIVE: To investigate the antioxidant and hepatoprotective properties of the different extracts Guizhencao (Herba Bidentis Bipinnatae) against liver injury in hyperlipidemia rats. METHODS: The rats were divided into 7 groups, with 10 rats in each. Rats were treated with high-fat diet for 18 weeks besides the normal control group, then rats in both normal control and model groups were received 5 mL/kg -1 · day -1 of saline and those in the positive control group with 2 mg/kg -1 · day -1 of lovastatin. Rats in the positive control group and different Guizhencao (Herba Bidentis Bipinnatae) extracts treatment groups (ethyl acetate extract group, n-hexane extract group, ethanol extract group, and aqueous extract group) were treated with corresponding extract at a concentration of 5 mL/kg -1 · day -1 . After 8 weeks treatment, all rats were sacrificed and total blood samples were collected. Histological analysis of liver was underdone by hematoxylin and eosin. The levels of serum ala-nine aminotransferase (ALT), aspartate aminotransferase (AST), glouse (GLU), cholesterol (TC), triglycerides (TG), low-density lipoprotein-cholesterol (LDL-C) and high-density lipoprotein-cholesterol were measured according to standard procedure using auto-analyzer. The superoxide dismutase (SOD) and malondialdehyde (MDA) levels in liver were ananlyzed by procedure instruction. RESULTS: The histopathological analysis implied that the administration of Guizhencao (HerbaBidentis Bipinnatae) extracts resulted in hepatoprotective role compared with that of the model group. In addition, the high-fat diet caused a remarkable increase of ALT, AST, GLU, TC, TG, LDL-C and MDA levels. A decline in HDL-C and SOD concentrations and a reversal of effects were observed in different Guizhencao (Herba Bidentis Bipinnatae) extracts groups, especially in the aqueous extract and ethanol extract groups. CONCLUSION: The different extracts of Guizhencao (Herba Bidentis Bipinnatae) can play a protecting role against liver injury in hyperlipidemia rats maybe through decreasing ALT, AST, GLU, TC, TG, LDL-C and MDA levels and enhancing the liver anti-oxidative ability.