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 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.

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 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.

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 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 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.

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

Artificial blood is a type of liquid preparation with oxygen-loading capacity and can temporarily substitute some function of blood.The developed artificial blood can be divided into four categories:artificial synthetic hemo-globin,artificial red blood cells made from natural hemoglobin,perfluorocarbons,and stem cell-differentiated red blood cells.This review focuses on the domestic and foreign research progress of artificial blood in recent years,and discusses its clinical application value,development trend,and future research,in order to provide new ideas to the development the artificial blood products and promote clinical application.

Objective:To explore the clinical effect of observation and psychological intervention, splint or brace fixation as well as surgical treatment on habitual dislocation of the hip (HDH) and to combine the authors' data with a compilation of the cases from the literature, evaluingating the epidemiological characteristics of HDH and the treatment scheme to maintain the stability of hip joint by systematic literature review in children.Methods:A retrospective analysis of the relevant data of 11 patients (12 hips) with HDH were treated from March 2007 to March 2021, including 2 boys and 9 girls. The age of the first dislocation was 2.25 (1.66, 3.75) years old and 4.33 (3.33, 6.17) years old at the age of diagnosis. At the same time, the relevant literature reports were searched from 1932 to 2022, and 24 HDH patients reports and clinical studies were confirmed to be included in this study according to the inclusion and exclusion criteria. The data of 33 patients (38 hips) with HDH who were obtained in the literature, including 5 boys and 28 girls. The age of the first dislocation was 2.00 (1.50, 2.00) years old and 4.00 (2.55, 5.00) years old at the age of diagnosis. A total of 44 patients (50 hips) with HDH who were analyzed, including gender, age of first dislocation, age of diagnosis, mode of stimulating dislocation, side and direction of dislocation, accompanying symptoms, family history, trauma history and imaging examination. The data of 41 patients with HDH (3 patients were excluded due to lack of treatment description) were treated with observation and psychological intervention in 19 patients, splint or brace fixation in 13 patients, and surgical treatment in 9 patients. The femoral neck-stem angle, acetabular index, central edge angle (CE angle) and Reimers instability index were measured by AutoCAD software, and the hip function was evaluated by Harris standard. Meantime, the disappearance time of dislocation was recorded.Results:The average follow-up time of 44 patients (50 hips) with HDH were 4.05±2.93 years old, and the time of disappearance of dislocation after treatment were 1.28±1.19 years old. Patients were diagnosed as 7 boys and 37 girls, 30 right and 8 left as well as 6 bilateral, the age of the first dislocation was 2.64±1.54 years old and 4.52±2.64 years old at the age of diagnosis. The data of 39 patients took hip flexion, adduction, internal rotation or hip flexion and adduction as the mode of stimulating dislocation. There were 43 patients with posterior dislocation, 41 patients with an audible "click" sound during dislocation, 36 patients with painless dislocation, and 37 patients with "vacuum phenomenon" were captured at the moment of dislocation. All patients with HDH had no specific family history and obvious history of trauma. There was no significant difference in general data between observation and psychological intervention group, splint or brace fixation group as well as surgical treatment group ( P>0.05). Harris standard to evaluate hip function, CE angle and Reimers instability index of the affected side were significantly different from those before and after treatment at the moment of dislocation ( H=127.51, P<0.001; H=55.70, P<0.001; H=54.69, P<0.001). Compared with the immediate disappearance of dislocation in the surgical treatment group, the disappearance time of dislocation in the observation and psychological intervention group and the splint or brace fixation group were significantly longer, and the difference was statistically significant ( H=20.83, P<0.001). Conclusion:Without specific family history and obvious trauma at young girls, painless posterior dislocation of hip with an audible "click" sound and "vacuum phenomenon" at the moment of dislocation are the significant epidemiological characteristics of HDH. Observation and psychological intervention, splint or brace fixation are recommended as the initial treatment scheme. When conservative treatment is ineffective, surgical treatment is needed to stabilize the hip joint quickly.