A 460 bp internal fragment of the AcrA gene from Vibrio alginolyticus strain HY9901 was amplified by PCR with designed primers and the unknown flanking sequence of 5 '- and 3 '- ends of the AcrA gene was finally characterized by inverse PCR and nested PCR. Sequence analysis showed the AcrA gene contained 1101 bp ORF encoding 366 amino acids and the deduced amino acid sequence of the precursor from Vibrio alginolyticus strain HY9901 showed significant homology with the putative protein of other Vibrio species. The AcrA shows 76%, 73%, 71% and 70% homology with V.vulnificus strain YJ016, V. parahaemolyticus strain RIMD 2210633, V. splendidus strain 12B01 and V. cholerae O1 biovar eltor str. N16961 respectively.

Objective To compare the effect of application of autologous iliac alone and NovaBone combined with autologous iliac in the acetabular osteotomy of children with developmental dislocation of the hip (DDH).Methods Data of 113 cases of children with DDH who had undergone open reduction and acetabular osteotomy surgery from 2007 to 2011 were retrospectively analyzed.According to bone material using after acetabular osteotomy,the patients were divided into autogenous iliac bone graft group (52 cases,60 hips) and NovaBone combined with autologous iliac bone graft group (61 cases,67 hips).There were no statistical differences in gender,age,side,dislocation type,osteotomy and acetabular index between the two groups.The patients were evaluated by Lane's scoring criteria,Severin standards and McKay standards at 6 weeks,3 months,6 months,1-year and 2-year post-operation follow-up.The bone healing of acetabular osteotomy zone,radiography and function of hip were compared.Results 6 weeks and 3 months after operation,Lane bone healing score in NovaBone combined with autologous iliac bone graft group (6.4±1.3 points and 9.6±1.7 points respectively) was obviously superior to autogenous iliac bone graft group (4.7±1.5 points and 7.8±1.2 points respectively).And 6 months and 1 year after operation,the two groups were basically reached bone healing.The Severin standard results showed that the rate (94％,63/67) of Excellent and Good in NovaBone combined with autologous iliac bone graft group (excellent:41 hips,good:22 hips,Fair:4 hips) was significantly higher than the rate (83.3％,50/60) in autogenous iliac bone graft group (excellent:28 hips,good:22 hips,Fair:10 hips).The rate (16.7％,10/60) of Fair in autogenous iliac bone graft group was significantly higher than the rate (6.0％,4/67) in NovaBone combined with autologous iliac bone graft group.McKay standard results were consistent with the results of radiological evaluation.Conclusion As a novel bone defect repair material,NovaBone can promote early bone healing process of acetabular osteotomy areas.It also can improve the resistance of osteotomy area.NovaBone can get a satisfied result in acetabular osteotomy in children with DDH.

Objective To evaluate the role of spinal neuronal Mas-related gene receptor C (MrgC) in the maintenance of bone cancer pain (BCP) in mice.Methods A total of 132 SPF male C3H/HeJ mice, aged 8-10 weeks, weighing 18-22 g, were randomly divided into 4 groups (n=33 each) using a random number table: sham operation group (S group) , BCP group, bovine adrenal medulla peptide 8-22 (BAM8-22, a highly selective MrgC agonist) group (group BAM), and MrgC antibody group (group MA).BCP was produced by injecting α-MEM 20 μl containing 2×105NCTC2472 cells into the distal medullary cavity of right femur bone.While α-MEM 20 μl was injected only in group S.The artificial cerebrospinal fluid 5 μl was injected intrathecally in S and BCP groups, and BAM 8-22 8 nmol/5 μl and MrgC antibody 5 μl were injected intrathecally in BAM and MA groups, respectively, once a day for 7 consecutive days starting from the day 14 after inoculation of the tumor cells.At 1 day before inoculation (T0), before administration (T1) , and at 14, 16 19 and 21 days after inoculation (T2-5, at 0.5 h before the initial administration and 2 h after each administration) , the number of spontaneous flinches (NSF) and mechanical paw withdrawal threshold (MWT) were measured.Five animals selected from each group at each time point were sacrificed, and the lumbar enlargement segments of the spinal cord were removed for determination of MrgC expression in the spinal neurons (by immunofluorescence).Results Compared with group S, NSF was significantly increased, MWT was decreased, and the expression of MrgC was up-regulated at T1-5 in BCP, BAM and MA groups.Compared with group BCP, NSF was significantly decreased, MWT was increased, and the expression of MrgC was up-regulated at T2-5 in group BAM, and NSF was significantly increased, MWT was decreased, and the expression of MrgC was down-regulated at T2-5 in group MA.Conclusion Spinal neuronal MrgC is involved in the maintenance of BCP in mice.

BACKGROUND: With the rapid development of tissue engineering, a single biological scaffold material is hard to meet the needs of tissue engineering. Therefore, composite scaffolds with excellent performance will be obtained by combining two or more kinds of materials.OBJECTIVE: To detect the adherence and proliferation of dental pulp stem cells on the Chitosan-fibrin composite scaffold.METHODS: Dental pulp stem cells were isolated and extracted from C57 neonatal rats through modified enzyme-digestion method, and subcultured to the third generation, followed by adipogenic and osteogenic induction in vitro. Then, induced cells were identified. The chitosan-fibrinogen composite scaffold was prepared, and the pore size and porosity were determined. The chitosan-fibrin composite scaffold was co-cultured with passage 3 dental pulp stem cells to observe the cell proliferation by MTT assay, and the morphology of the composite scaffold, cell adhesion,proliferation and extracellular matrix secretion were observed under scanning electron microscope. In addition, the cells were inoculated directly on the bottom of culture plate as controls.RESULTS AND CONCLUSION: The dental pulp stem cells were successfully isolated and cultivated, and positive for osteogenic and adipogenic differentiation. The pore size and porosity of the composite scaffold was (105.32±22.10) μm and (87.714±1.276)%, respectively. The S-shaped proliferation curve in the experimental group was similar with that in the control group; the proliferation rate in the experimental group was significantly higher than that in the control group after 4-8 days of culture (P < 0.05). At the 2nd day after co-culture, the cells adhered tightly and grew well onto the composite scaffold; at the 4th day, enlarged cells began to proliferate obviously with abundant extracellular matrix; the surface and pores of the scaffold were full of cells at the 6th day. These results indicate that the chitosan-fibrin composite scaffold is suitable for the adhesion and proliferation of dental pulp stem cells.

Objective To explore absorption kinetics of roxatidine acetate hydrochloric (ROX) in intestine of rats.Methods The absorption kinetics and permeability of ROX under different concentrations and different intestinal segments were investigated by double wavelength spectrophotometry via the in situ perfusing method in rats.Results There was no significant difference in Ka of ROX under different concentrations.The absorption rate in rats descended in order of duodenum,jejunum,ileum and colon [(3.87±0.12)×10-2,(2.53±0.18)×10-2,(1.43-±0.10)×10-2,(0.91±0.15)×10-2 · h-1].Conclusion The absorption of ROX in intestine complies with the passive transport mechanism and first order kinetics.ROX is well absorbed in thewhole intestine.

Objective To report our method to determine lateral needle insertion point (intersection point skin marker) in closed reduction and percutaneous Kirschner wire fixation for pediatric supracondylar fracture of humerus.Methods From May 2012 to June 2014,125 children with supracondylar fracture of humerus were treated with closed reduction and percutaneous Kirschner wire fixation.Of them,60 did not use the intersection point method to determine the lateral needle insertion point (group A),including 46 boys and 14 girls,with an average age of 6.3 ±0.6 years.According to Gartland classification,28 cases were type Ⅱ and 32 cases type Ⅲ.In the other 65 children,the intersection point method was used to determine the lateral needle insertion point (group B),including 50 boys and 15 girls,with an average age of 6.4 ± O.9 years.According to Gartland classification,29 cases were type Ⅱ and 36 cases type Ⅲ.The 2 groups were compared in terms of operation time,fluoroscopy times,hospital stay and hospitalization cost.The efficacy was evaluated at the final follow-ups using Flynn criteria.The 2 groups were compatible without significant differences in preoperative general data (P ＞ 0.05).Results All the 125 children obtained successful closed reduction and percutaneous pin fixation,and an average follow-up of 13 months (from 12 to 15 months)as well.There were significant differences between groups A and B in average operation time (23.1 ± 15.3 min versus 17.5 ± 10.3 min) and fluoroscopy times (9.2 ± 1.0 times versus 5.3 ± 1.3 times) (P ＜ 0.05).There was no statistically significant difference between the 2 groups either in Flynn excellent to good rate [(98.3％ (59/60) versus 98.5％ (64/65)] (P ＞ O.05).Needle irritation occurred in 2 cases and Kirschner wire shift in one in group A while tensile blistering occurred in one in group B.Conclusion In closed reduction and percutaneous Kirschner wire fixation for pediatric supracondylar fracture of humerus,the intersection point skin marker is a simple and reliable method to determine the lateral needle insertion point,leading to considerable reduction in radiographic exposure for both patients and doctors.

OBJECTIVE:To optimize the formulation of Roxatidine acetate hydrochloride(ROX)sustained-release tablets. METHODS:ROX sustained-release tablets were prepared by direct powder compression method. Central composite design-response surface methodology was used to optimize the formulation with composite index of 1,4,8 h in vitro accumulative release rate as index,using mass ratio of lactose/microcrystalline cellulose(MCC)(m/m),ethyl cellulose(EC)amount and HPMC amount as factors. Validation test was also conducted. RESULTS:The optimal formulation was as follows as ROX 75 mg,lactose 45 mg, MCC 91 mg,EC 65 mg,HPMC 124 mg,micropowder silica gel 2 mg. 1,4,8 h in vitro accumulative release rates of prepared sustained-release tablets were(30.7 ± 0.5)%,(65.8 ± 0.7)%,(89.4 ± 0.6)%,respectively. Related errors of them to predicted value were 0.6%,0.8%,1.2%,respectively. CONCLUSIONS:ROX sustained-release tablets are prepared successfully,and sustained-release effect is consisted with the expected effect.

Objective This study was designed to investigate the prognostic implications of the intertumoral heterogeneity of molecular phenotype in multifocal and multicentric breast cancer ( MMBC ) . Methods The clinical and follow-up data of 146 patients with MMBC from Jan.2009 to Dec.2009 treated in Tumor Hospital Affiliated to Zhengzhou University were retrospectively analyzed .We used Kaplan-Meier curves to compare the survivals of patients who had tumors with molecular phenotypic heterogeneity and patients who had multifocal homogeneous tumors in molecular phenotype , and the survivals of patients who had heterogeneous tumor type and grade and who had homogeneous tumor type and grade .The corresponding hazard ratio was calculated by Cox proportional-hazards regression .Results Intertumoral heterogeneity in histological type and grade of multiple breast cancer was detected in 16 of 146 patients (11.0%) and in 10 of 146 patients( 6.8%), respectively.Interfocal heterogeneous molecular phenotype of multiple breast cancer was detected in 24 of 146 patients ( 16.4%) .There was no significant difference in 5-year disease-free survival in multifocal cancer patients who had heterogeneous histological type and grade and who had homogeneous type and grade tumors (75.0%vs.77.3%, P=0.808).Multifocal cancers patients who had heterogeneous tumorsin molecular phenotype compared with those with homogeneous tumors in molecular phenotype had worse 5-year disease-specific survival (78 .7%vs.58.3%,P =0.037) , and had a greater risk of recurrence( HR=2.130, 95%CI=1.027-4.420; P=0.042).Phenotyping the additional cancer foci influenced the therapeutic decision in up to 16 patients (11 .0%) .Conclusoi ns Multifocal breast cancer patients who had heterogeneous tumors in molecular phenotype have a statistically significantly shorter disease-free survival.Phenotyping the additional cancer foci and managing with proper therapeutic decision may reduce the risk of recurrence or metastasis , and improve the outcomes of the patients .

Objective This study was designed to investigate the prognostic implications of the intertumoral heterogeneity of molecular phenotype in multifocal and multicentric breast cancer ( MMBC ) . Methods The clinical and follow-up data of 146 patients with MMBC from Jan.2009 to Dec.2009 treated in Tumor Hospital Affiliated to Zhengzhou University were retrospectively analyzed .We used Kaplan-Meier curves to compare the survivals of patients who had tumors with molecular phenotypic heterogeneity and patients who had multifocal homogeneous tumors in molecular phenotype , and the survivals of patients who had heterogeneous tumor type and grade and who had homogeneous tumor type and grade .The corresponding hazard ratio was calculated by Cox proportional-hazards regression .Results Intertumoral heterogeneity in histological type and grade of multiple breast cancer was detected in 16 of 146 patients (11.0%) and in 10 of 146 patients( 6.8%), respectively.Interfocal heterogeneous molecular phenotype of multiple breast cancer was detected in 24 of 146 patients ( 16.4%) .There was no significant difference in 5-year disease-free survival in multifocal cancer patients who had heterogeneous histological type and grade and who had homogeneous type and grade tumors (75.0%vs.77.3%, P=0.808).Multifocal cancers patients who had heterogeneous tumorsin molecular phenotype compared with those with homogeneous tumors in molecular phenotype had worse 5-year disease-specific survival (78 .7%vs.58.3%,P =0.037) , and had a greater risk of recurrence( HR=2.130, 95%CI=1.027-4.420; P=0.042).Phenotyping the additional cancer foci influenced the therapeutic decision in up to 16 patients (11 .0%) .Conclusoi ns Multifocal breast cancer patients who had heterogeneous tumors in molecular phenotype have a statistically significantly shorter disease-free survival.Phenotyping the additional cancer foci and managing with proper therapeutic decision may reduce the risk of recurrence or metastasis , and improve the outcomes of the patients .

OBJECTIVETo evaluate the sensitivity and specificity of body mass index (BMI), waist circumference (WC) and waist-to-hip ratio (WHR) measurements in diagnosing abdominal visceral obesity.

METHODSBMI, WC, and WHR were assessed in 690 Chinese adults (305 men and 385 women) and compared with magnetic resonance imaging (MRI) measurements of abdominal visceral adipose tissue (VA). Receiver operating characteristic (ROC) curves were generated and used to determine the threshold point for each anthropometric parameter.

RESULTS1) MRI showed that 61.7% of overweight/obese individuals (BMI > or = 25 kg/m2) and 14.2% of normal weight (BMI < 25 kg/m2) individuals had abdominal visceral obesity (VA > or = 100 cm2). 2) VA was positively correlated with each anthropometric variable, of which WC showed the highest correlation (r = 0.73-0.77, P < 0.001). 3) The best cut-off points for assessing abdominal visceral obesity were as followed: BMI of 26 kg/m2, WC of 90 cm, and WHR of 0.93, with WC being the most sensitive and specific factor. 4) Among subjects with BMI > or = 28 kg/m2 or WC > or = 95 cm, 95% of men and 90% of women appeared to have abdominal visceral obesity.

CONCLUSIONMeasurements of BMI, WC, and WHR can be used in the prediction of abdominal visceral obesity, of which WC was the one with better accuracy.

Abdomen ; Adipose Tissue ; Adult ; Aged ; Body Composition ; Body Mass Index ; China ; Female ; Forecasting ; Hip ; Humans ; Male ; Middle Aged ; Models, Theoretical ; Obesity ; Reference Values