Hapten-carrier protein conjugates were made using ciprofloxacin (CPFX) and two carrier proteins by 1-ethyl-3-(3- dimethylaminopropyl)-carbodiimide hydrochloride (EDC) method. Ultraviolet spectrophotometry were used to demonstrated that the molecule conjugate ratio of CPFX to ovalbumin (OVA) and bovine serum albumin (BSA) are 6:1 and 13:1 respectively. Nondenaturing gel electrophoresis results revealed that the conjugate band migrates differently from that of the carrier protein alone and of the EDC-treated protein when as few as 6 molecules of CPFX are attached to the carrier protein. The results indicate that nondenaturing gel electrophoresis and ultraviolet spectrophotometry can be employed to analyze the molecule coupling ratio of CPFX to carrier proteins qualitatively and quantitatively.

AIM: To study the protective effects of intravenous (iv) CGRP on myocardial injury in rat. METHODS: Establish a rat myocardial ischemic injury model by subcutaneous injection of single dose of isoproterenol (ISO), and treat the model with single dose of iv CGRP. Two hours later, serum CK, LDH, MDA and SOD levels were measured, MDA and SOD in myocardial tissue were tested, and myocardial tissue structures were observed. RESULTS:(1) Serum MDA and tissue MDA levels increased significantly and serum SOD and tissue SOD decreased significantly in injury group, in the CGRP treated group, the above changes were reversed (P

OBJECTIVETo detect the heterogeneity of mitochondrial DNA (mtDNA) in black and white hair of patients with type 2 diabetes.

METHODSMtDNA was extracted from the hair shaft of the patients to amplify two target DNA fragment from mtDNA coding region and control region using PCR. The differences in the heterogeneity in the target DNA fragment was analyzed between diabetic patients and the control group with denaturing high-performance liquid chromatography (DHPLC).

RESULTSIn the control subjects and diabetic patients, the mtDNA heterogeneity in the black hair was 3% and 10% in 20-45 year-old groups and 9% and 17% in 45-70 year-old groups, as compared to 9%, 20%, 21%, and 40% in the white hair, respectively. The mtDNA heterogeneity in the black and white hair was both higher in the diabetic patients than in the control subjects of the same age group, and was also higher in older age subgroups in both control and diabetic groups (P<0.05). The white hair mtDNA showed a significantly higher heterogeneity than the black hair mtDNA in the two age groups of diabetic patients and in 45-70 year-old control group (P<0.05).

CONCLUSIONThe mtDNA heterogeneity in the hair increases in type 2 diabetic patients and show an association with aging.

Adult ; Age Factors ; Aging ; genetics ; Chromatography, High Pressure Liquid ; methods ; DNA, Mitochondrial ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; metabolism ; Female ; Genetic Heterogeneity ; Hair ; metabolism ; Humans ; Male ; Middle Aged ; Young Adult

Objective:To investigate the correlation between 131I uptake and therapeutic efficacy in metastatic differentiated thyroid carcinoma (DTC). Methods:The clinical data of 138 patients with metastatic DTC (42 males, 96 females, age range: 8-74 years) treated with 131I in nuclear medicine departments of 31 centers all over China were retrospectively analyzed. The lesional 131I uptake was quantitatively analyzed with target-to-nontarget (T/NT) ratio through the regions of interest in metastatic lesions confirmed by either planar or tomographic 131I SPECT/CT imaging. The efficacies of 131I treatment on the metastatic DTC were divided into complete remission (CR), partial remission (PR), stable disease (SD) and progress disease (PD) based on the change of the lesion diameter before and after the treatment. Factors which may affect therapeutic efficacy were assessed by the univariate (Kruskal-Wallis rank sum test, χ2 test and one-way analysis of variance) and binary logistic regression analyses. The receiver operating characteristic (ROC) curve of lesional T/NT ratio to predict the ineffectiveness of 131I therapy was performed. Results:A total of 1 165 efficacies were evaluated. The planar imaging results ( n=653) showed that there was no statistically significant difference of T/NT ratio among CR, PR, SD and PD groups ( χ2=4.15, P>0.05). The tomographic imaging results ( n=512) suggested CR, PR, SD and PD in 7.6%(39/512), 65.8%(337/512), 22.9%(117/512), and 3.7%(19/512) of individuals, respectively, and the T/NT ratio among the four groups was significantly different ( χ2=30.46, P<0.01). The univariate analysis also showed that age, stimulated thyroglobulin(sTg), 131I dose were the factors affecting therapeutic efficacy ( F or χ2 values: 2.561, 7.095 and 8.799, all P<0.05). Furthermore, binary logistic regression analysis revealed that older patients (odds ratio ( OR)=1.034, P=0.022) or patients with lower lesional T/NT ( OR=1.086, P=0.006) had a higher probability of ineffectiveness. The area under ROC curve for T/NT ratio to predict ineffectiveness was 0.726, and the cut-off value was 6.2, with a sensitivity of 78.7%(107/136) and a specificity of 73.1%(275/376). Conclusions:131I therapy is an effective treatment for metastatic DTC. The age at the time of metastatic diagnosis and the lesional T/NT ratio are independent influential factors for ineffectiveness of 131I therapy. When the leisonal T/NT ratio is lower than 6.2, the inefficiency of 131I is higher.

Objective To investigate the influence of different cell structures on the static and dynamic mechanical performance of porous titanium alloy scaffolds,and to provide a theoretical mechanical basis for the application of scaffolds in the repair of mandibular bone defects.Methods Porous titanium alloy scaffolds with diamond,cubic,and cross-sectional cubic cell structures were manufactured using three-dimensional printing technology.Uniaxial compression tests and ratcheting fatigue with compression load tests were conducted to analyze the static and dynamic mechanical performances of scaffolds with different cell structures.Results The elastic moduli of the diamond cell,cross-sectional cubic cell,and cubic cell scaffolds were 1.17,0.566,and 0.322 GPa,respectively,and the yield strengths were 71.8,65.1,and 31.8 MPa,respectively.After reaching the stable stage,the ratcheting strains of the cross-sectional cubic,diamond,and cubic cell scaffolds were 3.3%,4.0%,and 4.5%,respectively.The ratcheting strain increased with increasing average stress,stress amplitude,and peak holding time,and decreased with increasing loading rate.Conclusions The evaluation results of the static mechanical performance showed that the diamond cell scaffold was the best,followed by the cross-sectional cubic cell scaffold and the cubic cell scaffold.The evaluation results of the dynamic mechanical performance showed that the cross-sectional cubic cell scaffold performed the best,followed by the diamond cell scaffold,whereas the cubic cell scaffold performed the worst.The fatigue performance of the scaffold is affected by the loading conditions.These results provide new insights for scaffold construction for the repair of mandibular bone defects and provide an experimental basis for further clinical applications of this scaffold technology.

The small molecule nutrients and cell growth factors required for the normal metabolism of chondrocyte mainly transport into the cartilage through free diffusion. However, the specific mass transfer law in the cartilage remains to be studied. In this study, using small molecule rhodamine B as tracer, the mass transfer models of cartilage were built under different pathways including surface pathway, lateral pathway and composite pathway. Sections of cartilage at different mass transfer times were observed by using laser confocal microscopy and the transport law of small molecules within different layers of cartilage was studied. The results showed that rhodamine B diffused into the whole cartilage layer through surface pathway within 2 h. The fluorescence intensity in the whole cartilage layer increased with the increase of mass transfer time. Compared to mass transfer of 2 h, the mean fluorescence intensity in the superficial, middle, and deep layers of cartilage increased by 1.83, 1.95, and 3.64 times, respectively, after 24 h of mass transfer. Under lateral path condition, rhodamine B was transported along the cartilage width, and the molecular transport distance increased with increasing mass transfer time. It is noted that rhodamine B could be transported to 2 mm away from cartilage side after 24 h of mass transfer. The effect of mass transfer under the composite path was better than those under the surface path and the lateral path, and especially the mass transfer in the deep layer of cartilage was improved. This study may provide a reference for the treatment and repair of cartilage injury.
Cartilage, Articular ; Rhodamines/pharmacology* ; Chondrocytes

【Objective】 Diabetic mice could show learning and memory dysfunction, and we aimed to investigate the effect of Sigma-1 receptor agonist, PRE-084, on neurons and cognitive impairment in mice with type 1 diabetes (T1DM). 【Methods】 Twenty mice with T1DM induced by streptozocin, aged 8-10 weeks, and 20 control mice (CON) were randomly divided into four groups (CON+Vehicle, CON+PRE-084, T1DM+Vehicle and T1DM+PRE-084). Mouse primary neurons were cultured in high glucose medium with PRE-084 and control solvent, respectively. The body weight, food and water intake, and fasting blood glucose level of mice in each group were detected and recorded. The learning and memory abilities of mice were detected by new object recognition experiment. The mitochondria-associated endoplasmic reticulum membrane (MAM) structure of neurons in hippocampal CA1 area of mice was detected by transmission electron microscope. And the expression levels of ATP and reactive oxygen species (ROS) in hippocampus of mice were detected by biochemical kit. Cell viability and ROS level of primary neurons were detected by CCK8 and cellular ROS kit. 【Results】 PRE-084 reduced the increase of body weight, food and water intake, and blood glucose caused by diabetes. PRE-084 significantly ameliorated the learning and memory impairment of the mice with T1DM, improved the changes of MAM structure in neurons of hippocampal CA1 area of diabetic mice, increased the level of ATP in hippocampus of diabetic mice, and decreased the increase of ROS expression in diabetic hippocampus and neurons under high glucose conditions. 【Conclusion】 Sigma-1 receptor agonist, PRE-084, could improve learning and memory impairment in the mice with T1DM, which might be related to the structural changes of MAM, the increase of ATP production, and the decrease of ROS production in hippocampal neurons.

Objective To study stress relaxation behaviors of cartilage scaffolds under different degradation cycles by using finite element analysis combined with theoretical models. Methods Based on the established degradation theoretical model, the elastic modulus of the scaffold was calculated under different degradation cycles. The finite element model of cartilage scaffolds was established and stress relaxation simulation was performed to analyze the variation of scaffold relaxation stress with time. The stress relaxation constitutive model was established to predict mechanical properties of the scaffold. Results The elastic modulus of cartilage scaffolds at 14 th, 28th, 42nd, 56th day after degradation was 32. 35, 31. 12, 29. 91, 28. 74 kPa, respectively. The upper layer for cartilage scaffolds was the largest. The overall relaxation stress of the scaffold decreased rapidly with time and then tended to be stable. At 8th week after degradation, the stress which the scaffold couldwithstand was still within the physiological load range of the cartilage. The predicted results of the stress relaxation constitutive model were in good agreement with the finite element simulation results. Conclusions The elastic modulus of the scaffold gradually decreases with the increase of degradation time. The longer the degradation period is, the less stress the scaffold can withstand. At the same degradation period, the larger the applied compressive strain, the larger the stress on the scaffold. Both the finite element simulation and stress relaxation constitutive model can effectively predict stress variations of cartilage scaffolds under degradation

Objective To study the effect of irrigation mechanical stimulation on scaffold degradation by numerical simulation, so as to predict its degradation degree. MethodsBased on perfusion experimental data, the fluid-solid coupling model was established by Comsol. The finite element model of scaffold was established by ABAQUS. Based on the models, the degradation performance of scaffold was simulated and predicted. Results The fluid-solid coupling simulation showed that the initial pressure at the speed of 15.79 mL/min was two-fold of that at 7.89 mL/min. Along the thickness of scaffold from the surface to the bottom, the pressures between the two velocities were decreased and gradually close to each other. The degradation of scaffold structure could be simulated dynamically by combining the degradation constitutive model with the finite element model. The obtained degradation data were consistent with the experimental data, and the residual molecular weight reached 0.643 on the 56th day. Compared with the experimental data, the simulation accuracy was higher than 98%. Conclusions The larger the perfusion velocity is, the greater the pressure on scaffold will be. Under the same perfusion velocity, the maximum force occurs on the surface of scaffold. The degradation pattern of scaffold can be predicted by applying the degradation constitutive model and the finite element model.

【Objective】 To explore the effect of high-fat and high-fructose diet on mouse intestinal barrier function, as well as the role of ketohexokinase (KHK), the key enzyme in fructose metabolism, in intestinal barrier impairment. 【Methods】 Eight-week-old male control C57BL/6J mice and Khk^-/- mice were randomly divided into control + normal diet (ND), control + high-fat and high-fructose diet (HFHFD), Khk^-/-+ normal diet (ND+Khk^-/-), and Khk^-/-+ high-fat and high-fructose diet (HFHFD+Khk^-/-) groups, with eight mice in each group. During the high-fat and high-fructose diet and normal diet, the body weight changes of mice in different groups were recorded. After the intervention, the blood glucose and insulin levels of mice in each group were detected. The intestinal barrier function and inflammation level of mice were evaluated by detecting intestinal water content, permeability, tight junction protein expression, serum and intestinal inflammatory factor levels. 【Results】 Compared with ND group, HFHFD group significantly increased the body weight, blood glucose and insulin levels of mice, increased the intestinal water content and permeability, decreased the expression of tight junction proteins, and increased inflammatory factors of the serum and intestines. In the two groups fed with high-fat and high-fructose diet, the body weight, blood glucose and insulin levels of the HFHFD+Khk^-/- group were significantly lower than those of HFHFD group, and the intestinal barrier dysfunction and inflammation were significantly improved. 【Conclusion】 KHK, a key enzyme in fructose metabolism, is involved in the impairment of intestinal barrier caused by high-fat and high-fructose diet. Knockout of Khk gene significantly improved intestinal barrier dysfunction and the inflammation level.