OBJECTIVETo optimize the hydrolysis process of linarin by response surface methodology, and to use the model of aldose reductase to study the acacetin's activity of aldose reductase inhibitory.

METHODThe model of acacetin enzyme in vitro was established by the determination of fluorescence absorption of NADPH, the inhibition rate of acacetin aldose reductase was calculated, and then the IC50 of hydrolysis was obtained. The hydrolysis process of linarin hydrolysis condition was optimized by using response surface method.

RESULTThe results indicated that the IC50 of acacetin (2.74 mg x L(-1)) was less than the IC50 of linarin (3.53 mg x L(-1)). Hydrolyzation time of 7.4 h, sulphuric acid concentration of 0.54 mol x L(-1) and the ratio of material to liquid of 3 : 1 were the optimum conditions.

CONCLUSIONHydrolyzate acacetin has preferable inhibitory activity of aldose reductase. The optimized hydrolysis condition of linarin is convenient to use with good predictability.

Aldehyde Reductase ; antagonists & inhibitors ; metabolism ; Animals ; Diabetic Retinopathy ; enzymology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Enzyme Inhibitors ; chemistry ; pharmacology ; Flavones ; chemistry ; Glycosides ; chemistry ; pharmacology ; Glycosylation ; Humans ; Hydrolysis ; Male ; Rats ; Rats, Wistar ; Retina ; enzymology

Congenital cataract is a highly heterogeneous disorder at both the genetic and the clinical-phenotypic levels. A unique cataract was observed in a 4-generation Chinese family, which was characterized by autosomal dominant inheritance and late-onset. Mutations in the 13 known genes (CRYAA, CRYAB, CRYBB1, CRYBB2, CRYGC, CRYBA1/A3, CRYGD, Connexin50, Connexin46, intrinsic membrane protein LIM2, cytoskeletal protein BFSP2, the major intrinsic protein-MIP and the heat shock factor HSF4) have previously been demonstrated to be the frequent reason for isolated congenital cataracts, but the exact molecular basis and underlying mechanisms of congenital cataract still remain unclear. This study was designed to find whether these 13 genes developed any mutation in the family members and to identify the disease-causing gene. Polymerase chain reaction (PCR) and direct DNA sequence analysis were carried out to detect the 13 genes. The results showed that no mutation causing amino acid alternations was found in these potential candidate genes among all patients in the family, and only several single-nucleotide polymorphisms (SNPs) were identified. A transitional mutation in the fourth intron of CRYBB2 and some silent mutations in the first exon of BFSP2 and CRYGD were found in the cataract family, but further study showed that these mutations could also be found in normal controls. It was concluded that some unidentified genes may underlie the occurrence of late-onset cataract in this family. A genome-wide screening will be carried out in the next study.

Objective To study the etiology of nosocomial infection and the risk factors in senile patients with acute myelogenous leukemia who received chemotherapy, then to explore the preventive measure. Methods 9lcases of senile patients with AML received 304 times of chemotherapy, then to analyze the rate of nosocomial infection, the infection site, the result of etiological examination and the relationship between the rate of nosocomial infection and the absolute of neutrophil count in peripheral blood and the duration of agranulocytosis. And then the results were compared with the non-senile patients with AML. Results The statistical indicators of nosocomial infection in senile patients with AML was higher than those younger ones. The rate of infection in lower respiratory tract, oral, alimentary canal, skin, anal area and the rate of sepsis in senile people was higher than those younger ones.Positive rate of fungal infection of senile patients was higher. The nosocomial infection in the patients whose absolute neutrophil count in peripheral blood was no more than 0.5×l09/L,and prolonged over 7 days was specific higher. Conclusions Senile patients with AML is the high risk group for nosocomial infection. Age, chemotherapy, the neutrocytopenia level and prolonged time are related to the nosocomial infection in senile patients with AML. It is necessary to take positive measure to control nosocominal infection and advance life quality of senile patients.

Objective To study the effect of hyperoxia solution and hypertonic saline infusion on arteries blood gas analysis of traumatic hemorrhagic shock rabbits,and interaction of hyperoxia solution and hypertonic saline.Methods Forty rabbits established into traumatic hemorrhagic shock model by Lamson' s method.Post hoc analysis was adopted in trial according to hyperoxia solution and hypertonic saline and 40 rabbits were divided into four groups(n =10):saline(NS) group,normal hyperoxic solution (NSO) group,hypertonic saline(HS) group,hyperosmotic-hyperoxic solution(HSO) group.Then blood samples were collected at pre-shock,post-shock,the end of limited resuscitation and hemostatic resuscitation for blood gas analysis.Results At the pre-shock,post-shock stages,PaCO2 and PaO2 were no significant difference in each groups ( P ＞0.05 ).At the end of limited resuscitation and hemostatic resuscitation stage,PaCO2 in NSO group (33.50 ±5.93),(37.22 ±6.74) mm Hg,(31.70 ±7.39),(35.10 ±7.56 )mm Hg in HS group,HSO group(38.70 ±4.92),(41.80 ± 5.51 )mm Hg were higher than that in NS group ( 27.40 ± 3.60 ),( 30.83 ± 2.79 ) mm Hg (F =6.894,4.287,respectively ;P ＜ 0.05 ),but there were no cross correlation between hypertonic saline and hyperoxia solution ( P ＞ 0.05).There were no significant difference in PaO2 in each groups at the end of limited resuscitation and hemostatic resuscitation stage ( P ＞ 0.05 ).Conclusions Both hyperoxia solution and hypertonic saline can effectively relieve hyperventilation,but there is no interaction between them.

Objective To explore the value of quantitative measurement of carotid intima-media thickness and elasticity for evaluating the degree of coronary heart disease by radio frequency ultrasonography. Methods From February 2011 to June 2012, a total of 142 subjects in cardiovascular internal medicine of the fourth military medical university Tangdu hospital with suspected CAD were enrolled in this study and were divided into non CAD group (n=35), atherosclerosis group in which coronary stenosis rate was less than 50%and no hemodynamic changes were detected (n=33), single artery involved group in which coronary stenosis rate was more than 50% (n=36) and multi-artery involved group in which coronary stenosis rate was more than 50%(n=38) based on the results of the coronary angiography (CAG). Measure the left common carotid artery intima-media thickness (IMT) and blood vessel elasticity index including the left common carotid artery pulse wave velocity (PWV) and stiffness index (β). Receiver operating characteristic (ROC) curves were used to calculate and compare the degrees of coronary heart disease, area under the curve of the left carotid artery elasticity parameters, cut-off value,sensitivity, speciifcity and Youden's index. Results LIMT, LPWV and Lβhad statistical signiifcance differences among four groups (F=3.621, 4.769 and 4.756, all P<0.05). The maximum values of parameter area under ROC curve were all observed in multi-artery involved groups and were 0.759 (LIMT), 0.770 (LPWV) and 0.764 (Lβ). The maximum value of Youden's index of the left common carotid artery intima-media thickness was observed in the coronary atherosclerosis group and was 0.513. The maximum value of Youden's index of the left common carotid artery pulse wave velocity and stiffnessβwere both observed in the multi-artery involved group and were 0.524 and 0.469. For diagnosing multivessel lesions, the sensitivities of LIMT, LPWV and Lβwere 71.0%, 77.4%and 71.0%, and the speciifcities were 73.4%, 74.7%and 75.9%. Conclusions The carotid elastic parameters have better predictive value for multivessel lesions. LPWV and Lβhave higher predictive value for coronary arterial disease than LIMT.

OBJECTIVETo explore the indications of arthroscope for the treatment of knee osteoarthritis and investigate the correlation between knee osteoarthritis imaging and effects of arthroscope.

METHODSFrom 2005.8 to 2008.4, 86 patients with knee osteoarthritis underwent arthroscope examination and treatment. Among the patients, 44 patients were male, and 42 patients were female, ranging in age from 46 to 67 years, averaged 56.3 years. Arthrodial cartilage of knee was graded by ICRS MR, and by Kellgern Laqrence X-ray. All the patients were followed up, and the duration ranged from 12 to 30 months. The Lysholm score was evaluated at the follow-up time.

RESULTSAmong 86 knees in 86 cases, cartilage injury degree of knees was graded as follows: grade 4 in 30 cases, grade 3 in 22 cases, grade 2 in 20 cases, grade 1 in 12 cases, grade 0 in 2 cases, mean grade (2.77 +/- 1.138). Postoperative Lysholm score ranged from 59 to 100, averaged (95.17 +/- 7.556), Kendall's correlation coefficient was -0.089, P = 0.317. There was no correlations between cartilage injury degree and Lysholm score. X-ray of knees was graded as follows: grade 4 in 0 cases, grade 3 in 24 cases, grade 2 in 38 cases, grade 1 in 17 cases, grade 0 in 7 cases, mean grade was (2.13 +/- 0.67), the Kendall's correlation coefficient was -0.851 with negative correlations (P = 0.036) between postoperative Lysholm score and K/L grade.

CONCLUSIONThere is no correlation between the grade of knee cartilage injury confirmed by MRI (1.5T) and effects of arthroscopy, and the grade is not a gold standard as an operation indication in arthroscope procedure. The K/L grade in X-ray had important effects.

Aged ; Arthroscopes ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Osteoarthritis, Knee ; diagnostic imaging ; pathology ; Radiography

OBJECTIVETo summarize clinical application of the distal island muscle flap of the gastrocnemius muscle for repairing the defects around knee and within the upper 2/3 of lower leg.

METHODS18 patients with defects around knee and within the upper 2/3 of lower leg, were treated with the distal island muscle flap of the gastrocnemius muscle, including 12 cases with medial muscle head and 6 cases with lateral muscle head. The muscular flaps were covered with meshed split-thickness skin grafts. The wound at donor sites were closed directly.

RESULTSAll the skin grafts and muscle flaps survived with superior infection at the wound edge in two cases. The patients were followed up for 8 months to 4.8 years with satisfactory cosmetic and functional results.

CONCLUSIONSThe distal island muscle flap of the gastrocnemius muscle has a reliable blood supply, long pedicle and suitable thickness. It is an ideal method for repairing the defects around knee and within the upper 2/3 of lower leg.

Adult ; Female ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Muscle, Skeletal ; transplantation ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the relationship between myeloperoxidase (MPO) 129 A/G promoter polymorphisms and the severity of coronary artery disease (CAD).

METHODSThe study enrolled 267 patients who had been diagnosed as coronary artery diseases by coronary angiography. The serum MPO activity was detected by colorimetric method. PCR-RFLP method was used to decide the genotypes of the patients. The severity of CAD was evaluated by the numbers of stenotic coronary arteries and the Gensini scores respectively.

RESULTSThe MPO 129 locus G and A alleles frequency were 0.893 and 0.107, respectively. No significant difference was observed in serum MPO activity between different genotypes (P > 0.05). The distribution of genotypes in different Gensini score groups had no significant difference (P > 0.05). The patients with GG genotypes were prone to develop mutivessel diseases.

CONCLUSIONNo significant correlation exists in MPO 129 locus polymorphism and serum MPO activity. The MPO 129 locus polymorphism isn't a reasonable predict factor of CAD severity.

Adult ; Aged ; Aged, 80 and over ; Coronary Angiography ; Coronary Artery Disease ; blood ; genetics ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Peroxidase ; blood ; genetics ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Risk Factors ; Severity of Illness Index

A continuous ethanol fermentation system composed of four-stage tank fermentors in series and with a total working volume of 4000 mL was established. The first fermentor was designated as the seed fermentor and the others for ethanol fermentation. A self-flocculating yeast strain developed by protoplast fusion of Saccharomyces cerevisiae and Schizosaccharomyces pombe was applied. Two-stage corn powder enzymatic hydrolyzate containing reducing sugar 100 g/L, together with 2.0 g/L (NH4)2HPO4 and KH2PO4, was used as yeast seed culture medium and fed into the seed fermentor at the dilution rate of 0.017h (-1). Meanwhile, the hydrolyzate containing reducing sugar 220 g/L, added with 1.5 g/L (NH4)2HPO4 and 2.5 g/L KH2PO4, was used as ethanol fermentation substrate and fed into the second fermentor at the dilution rates of 0.017, 0.025, 0.033, 0.040 and 0.050 h(-1) (based on the total working volume of the three fermentors), respectively. The chemostat states on which all of the monitoring parameters, including residual sugar, ethanol and yeast cell biomass concentrations, were maintained relatively constant were observed for seed cultivation and ethanol fermentations when the fermentation system was operated at the dilution rates of 0.017, 0.025, 0.033 and 0.050 h(-1). Yeast cells were observed being partly immobilized because significant yeast cell biomass concentration differences between the broth out of and inside the fermentors were detected. Moreover, the oscillations of residual sugar, ethanol and yeast cell biomass concentrations were observed when the fermentation system was operated at the dilution rate of 0.040 h(-1). The broth containing more than 12% (V/V) ethanol and less than 0.11% (W/V) residual reducing sugar and 0.35% (W/V) residual total sugar was produced when the dilution rate was controlled at no more than 0.033 h(-1). The ethanol productivity was calculated to be 3.32(g x L(-1) x h(-1)) for the dilution rate of 0.033 h(-1), which increased nearly 100% compared with that for conventional ethanol fermentation technologies using freely suspended yeast cells.
Bioreactors ; microbiology ; Carbohydrates ; Ethanol ; metabolism ; Fermentation ; Flocculation ; Saccharomyces cerevisiae ; metabolism ; Schizosaccharomyces ; metabolism ; Zea mays ; metabolism

The fusion protein of Humanized mouse anti-human fibrin ScFv and the low molecular weight urokinase (IIn-UK) contained seven disulfide bonds and formed inclusion body while expressing in normal E. coli strain. By coexpressing DsbC and using the special E. coli strain Origami(DE3) which was trxB/gor double mutant, the fusion protein IIn-UK was functionally expressed in the cytoplasm of E. coli. The expressed fusion protein in the soluble fraction was purified by using affinity chromatography specific against urokinase. The purified fusion protein could combine the thrombus in vitro, and the specific activity of urokinase reached 80,000 IU/mg fusion protein. The result showed that the fusion protein retained the activity of two moieties, and this study laid a foundation for further research of targeting thrombolytic agent.
Animals ; Chromatography, Affinity ; Escherichia coli ; genetics ; Gene Expression ; Humans ; Immunoglobulin Fragments ; genetics ; Mice ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Urokinase-Type Plasminogen Activator ; genetics