Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Adolescent ; Humans ; Linezolid ; therapeutic use ; Male ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; pathogenicity ; Pneumonia ; diagnostic imaging ; microbiology ; Vancomycin ; therapeutic use

Objective To detect the molecular characterization of polysaccharide purified from Amusium pleuronectes, so as to investigate its role of intervention to the formation of hepatic fibrosis caused by Schistosoma japonicum infection. Methods The crude polysaccharide from A. pleuronectes was extracted and further purified, and the molecular weight and monosaccharide composition were determined by the high pressure size exclusion chromatography and PMP pre-column derivatization method, respectively. A total of 50 female BALB/c mice were randomly divided into five groups：A (normal group), B (experimental group), C (polysaccharide group), D (praziquantel), and E (polysaccharide + praziquantel group). The mice in B, C, D, or E groups were attacked on the abdominal skin by using the cercariae of S. japonicum (30 ± 2 for each mouse) respectively. After 8 weeks, the mice in C, D, and E groups were administrated by polysaccharide and/or praziquantel, and the mice in B group were instead of saline. All the livers and sera were collected after 16 weeks. HE staining was employed for the livers, and serum IFN-γ and IL-13 were measured by using ELISA kits. Results The molecular weight of purified polysaccharide from A. pleuronectes was 11.7 kDa. Compared with A and B groups, the serum levels of IFN-γ in C, D, and E groups were significantly increased (F = 63.525, P < 0.01). However, the serum levels of IL-13 in C, D, and E groups were significantly decreased (F = 99.788, P < 0.01) compared with that in B group. HE staining showed that the egg nodules and hepatic fibrosis were observed in B, C, D, and E groups. The number of egg nodules and fibrosis degree in E group were milder than those in B group (χ² = 7.875, P < 0.05). Conclusions The polysaccharide from A. pleuronectes has an obvious effect in preventing hepatic fibrosis process induced by S. japonicum infection, particularly combining with the administration of praziquantel.

OBJECTIVETo explore the safety and feasibility of biodegradable magnesium alloy stapler based on the result of animal experimental study for gastrointestinal anastomosis.

METHODSSixteen beagle dogs were equally and randomly divided into experimental (magnesium alloy) group and control (titanium alloy) group. A gastrojejunal and a colonic anastomosis were performed in each beagle dog. The anastomosis time, postoperative complications, body weight, blasting pressure of anastomosis and serum glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, creatinine, blood urea nitrogen, and serum magnesium were compared between the two groups. The healing of anastomosis and degradation of magnesium alloy were observed. The histopathological features of heart, liver, spleen and kidney were examined in the two groups.

RESULTSThere were no significant differences in anastomosis time, body weight, postoperative complications, anastomotic bursting pressure between the two groups. The anastomosis was healed well, and no dramatic inflammatory cell infiltration was observed. Magnesium alloy could be degraded completely in the animal body within 90 days. There were no significant differences in serum glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, creatinine, blood urea nitrogen and serum magnesium between the two groups. Histopathological examination showed that the degradation of magnesium alloy did not harm the important organs (liver, kidney, heart, brain and spleen).

CONCLUSIONSMagnesium alloy stapler is safe and feasible for gastrointestinal anastomosis in beagle dogs. The degradation of magnesium alloy does not harm the healing of anastomosis and other important organs. Magnesium alloy stapler may be a candidate of biodegradable and safe material of stapler for gastrointestinal anastomosis in human.

Absorbable Implants ; Alloys ; Animals ; Dogs ; Female ; Gastroenterostomy ; instrumentation ; Magnesium ; Male ; Sutures ; Titanium

OBJECTIVETo evaluate the effect of maxillary sinus elevation with gene-enhanced tissue engineering bone in dogs.

METHODSbone marrow stromal cells (BMSC) derived from dog marrow were cultured, and transduced with the adenovirus carrying bone morphogenetic protein-2 (BMP-2) gene (AdBMP-2), the adenovirus carrying green fluorescent protein (GFP) gene (AdGFP) in vitro. The bone formation ability of gene modified BMSC with scaffold was examined in nude mice and in elevated maxillary sinus of dog. Student's t-test was used for statistical analysis with SPSS 11.0 software package.

RESULTSGene transfection efficiency reached up to (83.95 ± 2.43)% as demonstrated by GFP expression. Ectopic bone formation was detected in nude mice. As for maxillary sinus floor elevation in a dog model, new bone formation area in the AdBMP-2 gene transduced BMSC with Bio-Oss group was significantly higher than in BMSC with Bio-Oss group at 120 d (P < 0.05).

CONCLUSIONSAdBMP-2 gene transduced BMSC can stimulate ectopic bone formation in nude mice, and promote bone formation and maturation in the dog maxillary sinus.

Adenoviridae ; genetics ; Animals ; Bone Matrix ; transplantation ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Bone Transplantation ; methods ; Cells, Cultured ; Dogs ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Nude ; Minerals ; Osteogenesis ; Recombinant Proteins ; genetics ; metabolism ; Sinus Floor Augmentation ; methods ; Tissue Engineering ; methods ; Transfection

Objective To investigate the variance levels of plasma soluble leukocyte differentiation antigens CD40 (sCD40) and soluble CD40 ligand (sCD40L) in preeclamptic patients with renal damage and its relationship. Methods A total of 63 pregnant women attended the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College between August 2008 and June 2010. In the present study included 28 pregnant women with mild preeclampsia and 35 patients with severe preeclampsia. Thirty matched normotensive pregnant women were enrolled in the study as the control group. Expression of sCD40 and sCD40L were determined by ELISA. At the same time, the blood routine, C reaction protein ( CRP),urine routine, 24 hours urine protein excretion, and serum uric acid (UA), creatinine (Cr), blood urea nitrogen (BUN) were measured. The correlation analysis was performed between the sCD40/sCD40L and the blood biochemical indexes in 3 groups. Results ( 1 ) The median levels of CRP in severe preeclampsia (10. 8 mg/L)and mild preeclampsia group(7. I mg/L)are significantly higher than that of control group (3. 3 mg/L,P ＜ 0. 05 ); The level of CRP in severe preeclampsia group was also higher than that of mild preeclampsia group ( P ＜ 0. 05 ). The median gestational age at delivery in severe preeclampsia ( 32. 5 weeks)was significantly less than that of mild preeclampsia group ( 37. 2 weeks) and normal group ( 38. 6 weeks,P ＜ 0. 05). However no significant differences were observed between mild preeclampsia group and normal group ( P ＞0. 05 ). The platelet count in severe preeclampsia ( 132 × 109/L) was significantly less than those of mild preeclampsia group (212 × 109/L) and normal group ( 216 × 109/L, P ＜ 0. 01 ), but no significant differences were observed in blood platelet amount between mild preeclampsia group and normal group ( P ＞0. 05 ). There was no significant difference in hemoglobin level and white blood cell in three groups ( P ＞0. 05). (2) The sCD40 plasma concentration in severe, mild preeclampsia and normal group was 133.6,126. 5 and 90. 7 ng/L, respectively. The sCD40 L plasma concentrations were 12. 5, 10. 4 and 4. 4 ng/L respectively in the 3 groups. 24 hours urinary protein quantitative was 4. 5 g/d,0. 8 g/d and 0 in the 3 groups respectively. And the UA level was 486 μ mol/L,289 μmol/L and 162 μmol/L. In the above three groups,the monitoring indicators were significantly higher in women with severe preeclampsia group compared with mild preeclampsia and control groups (P ＜ 0. 01 ), and there were also higher in mild preeclampsia group than that in control groups ( P ＜ 0. 01 ). The level of plasma Cr ( 89 μmol/L) and BUN ( 5. 32 mmol/L) in severe preeclampsia group were higher than those of mild preeclampsia group (66 μmol/L and 4. 49mmol/L) and control group ( 57 μmol/L and 3.32 mmol/L, P ＜ 0. 05 ). There was no significant difference between mild preeclampsia group and normal group (P ＞ 0. 05 ). (3) The correlation analysis indicated that the level of sCD40 has a positive correlation with 24 hours urinary protein quantitative( r = 0. 434, P ＜ 0. 05 ),also significant positive correlation( r =0. 536,0. 528 ,P ＜ 0. 01 ) between the level of sCD40 and UA or CRP in women with preeclampsia. There was no significant correlation between the level of sCD40 and systolic blood pressure, diastolic blood pressure, delivery gestational age, Cr, BUN, and platelet count(r =0. 135,0. 183, -0. 133,0. 190,0. 167, -0. 221 ,all P ＞0. 05 ). There were positive correlation between the level of sCD40L and 24 hours urine protein excretion, either UA or CRP( r =0. 591,0. 445,0. 539 ,all P ＜0. 01 ). No significant correlation was found between sCD40 L and systolic blood pressure, diastolic blood pressure,delivery gestational age, Cr, BUN, and platelet count( r =0. 178,0. 212, -0. 292,0. 144,0. 135, -0. 273,all P ＞0. 05). There was significant positive correlation between plasma sCD40 and sCD40L ( r =0. 707 ,P ＜0. 01 ). There was no relationship between the level of sCD40, sCD40L and the blood biochemical indexes in normotensive pregnant women ( P ＞ 0. 05 ). Conclusions The plasma concentrations of sCD40 and sCD40 L are significantly higher in pregnant women with preeclampsia compared with the control, which may be involved in the development of preeclampsia and contribute to the kidney damage. The variance levels of sCD40 and sCD40L may be also related to the severity of preeclampsia.

Objective To investigate the expression of macrophage migration inhibitory factor (MIF) and CD_(74), the receptor of MIF, in preeclamptic placenta and its correlation with the pathogenesis of preeclampsia.Methods From March 2008 to November 2008,69 preeclamptic women who delivered in the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College,were recruited,including 33 women with mild preeclampsia (MPE group) and 36 women with severe preeclampsia (SPE group).Another 43 healthy pregnant women were taken as control group.Immunoturbidimetry was applied to measure the concentrations of C-reactive protein (CRP) in maternal blood.The expressions of MIF and CD_(74) in placenta were tested with immunohistochemistry and the expressions of MIF mRNA and CD_(74) mRNA were detected by semiquantitative RT-PCR.The relationship between maternal blood level of CRP and MIF mRNA and CD_(74) mRNA in placenta was analyzed in the MPE and SPE group.Results (1) MIF and CD_(74) were expressed in the placenta of all pregnant women in the 3 groups, as shown in brown-yellow color, and significantly higher expression was found in the MPE and SPE group.(2) The expression of MIF mRNA and CD_(74) mRNA in the MPE group (0.70±0.13 and 0.96±0.16), SPE group (0.88 ± 0.12 and 1.08 ± 0.15) were significantly higher than in the control group (0.67 ± 0.11 and 0.83 ± 0.14) (P < 0.01), and statistical significance was also found between the MPE and SPE group (P <0.01).(3)The maternal blood concentrations of CRP in the MPE and SPE group were significantly higher than in the control group [(15.3±7.0) mg/L and (21.6±9.1)mg/L vs (4.8 ± 1.8) mg/L, P <0.01] , and significant difference was also found between the MPE and SPE group (P <0.01).(4) In the two preeclamptic groups, the blood concentrations of CRP were positively correlated with the expression of both MIF mRNA(r =0.67 ,P <0.01)and CD_(74) mRNA(r =0.83 ,P <0.01) in placenta.Positive correlation was also found between the levels of MIF mRNA and CD_(74) mRNA in placenta (r =0.93 ,P < 0.01).Conclusions Overexpression of MIF and CD_(74) in the placenta may up-regulate the CRP level in maternal blood, resulting in systemic inflammatory reaction and vascular endothelium damage which may be involved in the pathogenesis of preeclampsia.

OBJECTIVETo evaluate the effect of tissue engineering bone in maxillary sinus lifting.

METHODSThe marrow stromal stem cells of dog were cultured in DMEM containing 100 m1/L fetal bovine serum and induced to differentiate to osteoblasts, which were then inoculated together with Bio-Oss for 5 days. Sixteen dog's bilateral maxillary sinus were elevated. One side was grafted with a compound of BMSC and Bio-Oss and the other side grafted with Bio-Oss alone. The samples were studied by gross, CT, histomorphology and histomorphometrical analysis at the 30th, 90th day after the operation. t-test was used for statistical analysis.

RESULTSIn gross view and CT, new bone formation was observed in all maxillary sinus after 30 and 90 days respectively. Histomorphometrical analysis showed much more new callus in BMSC-Bio-Oss group than in Bio-Oss group (P < 0.05).

CONCLUSIONSA better effect of new bone formation could be obtained with tissue engineered bone in maxillary sinus lifting.

Animals ; Bone Substitutes ; Cells, Cultured ; Dogs ; Maxillary Sinus ; surgery ; Minerals ; Myeloid Progenitor Cells ; transplantation ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo study the protective effects of Isodon lophanthoides var. gerardianus (ILVG) aqueous extract on the acute hepatic injury induced by carbon tetrachloride (CCl4) in rats.

METHODSixty rats were allocated into control group, model group, low, middle and high dosage group and Bifendate group randomly. At the test group, rats received either ILVG aqueous extract (15, 7.5, 3.75 g x kg(-1)) or Bifendate (45 mg x kg(-1)) by gastric perfusion daily for 10 consecutive days. In 1st, 4th, 7th and 10th days, 10% CCl4 (2 mL x kg(-1)) was given to rats by intraperitoneal (ip) injection. The rats were killed 24 h after the last adminiction with drug, the levels of ALT, AST, ALP and total bilirubin in serum were analyzed, the body weight, liver weight, spleen weight and thymus weight of each rat were measured, and the hepatic tissue pathology was observed.

RESULTILVG could decrease the ALT, AST, ALP and T-Bil in serum, restrain the enlargement of liver and the shrinkage of thymus, and reduce the necrosis in pathological observation.

CONCLUSIONILVG aqueous extract possesses the effects of protecting on the acute hepatic injury induced by CCl4 in rats.

Alanine Transaminase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; Carbon Tetrachloride Poisoning ; Chemical and Drug Induced Liver Injury ; blood ; etiology ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Isodon ; chemistry ; Liver ; pathology ; Male ; Plants, Medicinal ; chemistry ; Protective Agents ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley