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

AIMTo clarify the role of PTCH in patients with NBCCS-related and non-sydromic keratocystic odontogenic tumors.

METHODOLOGYMutation analysis was undertaken in 8 sporadic and 4 NBCCS-associated KCOTs.

RESULTSFour novel and two known mutations were identified in 2 sporadic and 3 syndromic cases, two of which being germline mutations (c.2179delT, c.2824delC) and 4 somatic mutations (c.3162dupG, c.1362-1374dup, c.1012 C>T, c.403C>T).

CONCLUSIONOur findings suggest that defects of PTCH are associated with the pathogenesis of syndromic as well as a subset of non-syndromic KCOTs.

Adolescent ; Adult ; Amino Acid Sequence ; Basal Cell Nevus Syndrome ; genetics ; Chromatography, High Pressure Liquid ; Codon, Nonsense ; genetics ; Codon, Terminator ; genetics ; Conserved Sequence ; genetics ; Cytosine ; Exons ; genetics ; Female ; Frameshift Mutation ; genetics ; Gene Duplication ; Germ-Line Mutation ; genetics ; Guanine ; Humans ; Male ; Middle Aged ; Mutation ; genetics ; Mutation, Missense ; genetics ; Odontogenic Tumors ; genetics ; Patched Receptors ; Patched-1 Receptor ; Receptors, Cell Surface ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Deletion ; genetics ; Syndrome ; Threonine ; genetics ; Thymine

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

OBJECTIVETo observe the effect of sensory neuropeptide substance P combined with epidermal stem cells (ESC) on wound healing and nerve regeneration in diabetic rats.

METHODSESC that had been isolated from SD rats were identified and cultured in vitro, and they were inoculated onto nourishing layer of amniotic membrane to construct amniotic membrane-ESC. Four full-thickness skin wounds were produced on the back of each of 48 diabetic rats. The resulted 192 wounds were randomly divided into ESC + substance P group, ESC group, substance P group, and control group according to the lottery method, with 48 wounds in each group. Wounds in ESC + substance P group and ESC group were transplanted with amniotic membrane-ESC, and those in substance P group and control group were transplanted with amniotic membrane. After transplantation, 250 µL substance P in the concentration of 1 × 10(-7) mol/L was injected around and into the middle of the wounds in ESC + substance P group and substance P group, 2 times a day, and continued for 4 days, while 250 µL PBS solution was injected in the above-mentioned position in ESC group and control group as control, 2 times a day, and continued for 4 days. On post injury day (PID) 4, 7, 10, 14, 17, and 23, the wound healing rate (with 8 wounds at each time point) was observed and determined, and changes in wound tissue structure were observed with HE staining. On PID 4, 7, and 10, collagen distribution in wound tissue was observed with Masson staining, and type I and type III collagen deposition in wound tissue was respectively observed after immunohistochemical staining. The distribution of protein gene product 9.5 (PGP 9.5) and regeneration of substance P positive nerve fibers in wound tissue were observed with immunohistochemical staining on PID 14 and 23. Data were processed with one-way analysis of variance and t test.

RESULTS(1) The wound healing rate in ESC + substance P group reached 100.0% on PID 14, which was obviously earlier than that in ESC group, substance P group, and control group, healing was respectively observed on PID 17, 17, and 23. The wound healing quality in ESC + substance P group was better than that in the other three groups as shown by HE staining. (2) On PID 10, collagen that was darkly stained and widely distributed was observed in wound tissue of ESC + substance P group and substance P group, while collagen in the other two groups was lightly stained and narrowly distributed. Deposition quantity of type I collagen gradually increased, and that of type III collagen gradually decreased in the wounds of each group over time. On PID 4, 7, and 10, distribution amount of type I collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.72, 118.21, 26.71, P values all below 0.01) and control group (with t value respectively 44.37, 22.76, 30.32, P values all below 0.01), while there was no significance between ESC + substance P group and substance P group. On PID 4, 7, and 10, distribution amount of type III collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.27, 28.68, 14.51, P values all below 0.01) and control group (with t value respectively 35.68, 22.52, 22.24, P values all below 0.01). (3) A large amount of PGP 9.5 and regeneration of substance P positive nerve fibers, and some peripheral nerve fibers in deep wound extending to epidermis were observed in wound tissue of ESC + substance P group and substance P group. A small amount of PGP 9.5 and regeneration of substance P positive nerve fibers without peripheral nerve fibers extending to epidermis were observed in deep wound tissue of ESC group and control group. On PID 14, 23, ratios of area of PGP 9.5 positive nerve fiber in the wounds of ESC + substance P group were (3.86 ± 0.25)% and (7.03 ± 0.28)%, and they were significantly higher than those of ESC group [(1.48 ± 0.30)%, (3.01 ± 0.43)%, with t value respectively 23.95, 30.27, P values all below 0.01] and control group [(1.46 ± 0.23)%, (2.84 ± 0.29)%, with t value respectively 27.35, 40.32, P values all below 0.01]. On PID 14, 23, ratios of substance P positive nerve fiber area in the wounds of ESC + substance P group were (2.01 ± 0.14)% and (1.19 ± 0.11)%, which were obviously higher than those of ESC group [(0.85 ± 0.17)%, (1.34 ± 0.21)%, with t value respectively 20.50, 2.60, P < 0.05 or P < 0.01] and control group [(0.74 ± 0.15)%, (1.30 ± 0.17)%, with t value respectively 23.98, 2.41, P < 0.05 or P < 0.01].

CONCLUSIONSJoint application of substance P and ESC can effectively promote healing of wound and nerve regeneration in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; pathology ; Epithelial Cells ; cytology ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; Substance P ; pharmacology ; therapeutic use ; Wound Healing