Objective To study the feasibility of microsurgical technique to denervate sympathetic of femoral artery in rabbit, providing a reliable animal experimental model for further study of the mechanism of neuralization in bone tissue engineering.Methods From July, 2014 to July, 2015, 21 New Zealand white rabbits were divided into 4 groups randomly: the control group (n =3), the 4 weeks group (n =6), the 8 weeks group (n =6) and the 12 weeks group (n =6).Bilateral femoral arteries of the 21 rabbits were exposed.Adventitia of femoral arteries in 3 test groups were removed for about 2cm by microsurgical technique, whereas adventitia of the control group remained intact without any treatment.The arteries samples were collected at 4 weeks, 8 weeks and 12 weeks after treatment.The structure of vascular were indicated by hematoxylin-eosin (HE) staining, and the distribution and volume of the sympathetic fibers were evidenced by glyoxylic acid staining and the expression of tyrosine hydroxylase (TH), the marked protein of sympathetic.Results The adventitia of 3 test groups were invisible or lost most of it while the control group remained intact shown by HE staining.For glyoxylic acid staining, the fluorescence intensity value of the control group, 4 weeks group, 8 weeks and 12 weeks were 0.08124 ± 0.00260, 0.02920 ± 0.00206, 0.02661 ± 0.00233, 0.03094 ± 0.00211, respectively (n =6).The distribution and fluorescence intensity of sympathetic nerve were both significantly reduced in test groups compared to the control group (P ＜ 0.05).And there was no statistical difference among the 3 test groups (P ＞ 0.05).Semi-quantitative analysis of the expression of TH was 0.8626 ± 0.03519, 0.3631 ± 0.03019, 0.3964 ± 0.02239, 0.3487 ± 0.02356 respectively, which showed the same tendency as glyoxylic acid staining test.Conclusion Microsurgical technique is promising as an ideal method for the local denervation of sympathetic nerve from artery system as it can significantly reduce sympathetic fibers on adventitia without regeneration during the experimental period.

Objective To study the conditions and methods of hydrodynamics-based transgene into rat regenerating liver in vivo. Methods The solution with concentration 30mg/L gene-containing plasmid was injected into rat tail veins at a speed of 2ml/s, then partial hepatectomy (PH) was performed at different times before/after injection, finally the rat (g) and regenerating liver (g) were weighed, and the liver coefficient (Lc) was calculated. Out of 15 groups which are Lc±Lc*0%, *5%, *10%, *15%, *20%, *25%, *30%, *35%, the most suitable group was chosen as correction coefficient to calculate the most appropriate volume of plasmid solution which was injected into the regenerating liver at different recovery times, and at the same time, right lobe of liver was gathered to make frozen section, then observe and quantify the positive green fluorescent protein (GFP) rate at 488 nm excitation wavelength. Results Injection of either physiological saline or empty plasmid has no significant difference compared with control (only PH performance). The appropriate time of hydrodynamics-based transgene is more than 12 hours before PH or anytime after PH. The solution volume of hydrodynamics-based transgene into liver regenerating rat after PH is rat weight (g) ×9%×1/3×corresponding correction coefficient (Trc). Both vector and target gene have effect on the time and abundance of gene expression. Conclusion Hydrodynamics-based transgene can effectively be applied to gene transfection in rat regenerating liver.

Objective To study the determinants of exercise behavior among the overweight and obese residents, so as to provide reference for policy-making and health promotion in the future. Methods The study only enrolled overweight and obese residents to conduct a questionnaire survey and physical examination including height, weight and waist circumference. The self-designed questionnaire based on Health Belief Model had acceptable validity and reliability. Structural Equation Modeling was used to analyze the determinants of exercise behavior after univariate analysis and correlation analysis. Result Totally 2 489 overweight and obese residents(1 182 male and 1 307 female)were included in this study. Their average age was 45.3±14.2 years and their median amount of exercise was 4 thousand-step equivalent with quartile of (0.5, 10.0); 22.9% of these residents never took any exercise. Males with high school and equivalent level of education and an income of 5 000 to 10 000 yuan, took more exercise than other demographic groups. Amount of exercise had a positive correlation with perceived benefits of exercise, perceived severity of disadvantage of low exercise and self-efficacy, while it had a negative correlation with subjective barriers and objective barriers. Structural Equation Modeling showed that perceived subjective barriers, objective barriers and self-efficacy were determinants of exercise behavior. Their total standardized effects on exercise behavior were-0.085 (P=0.020),-0.242 (P<0.001) and 0.184 (P<0.001) respectively.Those with higher self-efficacy and less barriers preferred to do more exercise. Indirect effect widely existed in the SEM and self-efficacy were the most important mediating variables of all the other significant determinants. Conclusion The overweight and obese residents are seriously lack of exercise, which need some targeted intervention, especially by improving their self-efficacy and reducing their objective barriers.

Objective To investigate the effect of prevascularized tissue-engineered bone graft on regeneration of femoral bone defects in rats.Methods Models of femoral bone defect were created at the bilateral hind limbs of 20 healthy female 10 week-old rats which were divided into 2 even groups randomly (n =10).In group A,conventional tissue-engineered bone grafts were transplanted into the femoral bone defects;in group B,tissue-engineered bone grafts and vascular bundles were implanted into the femoral defects.At 1,4 and 8 weeks after operation,3 rats were sacrificed each time in each group to harvest samples.The remaining one in each group served as a spare animal.Regeneration of bone defects and degradation of scaffolds were assessed by radiologic modality and hematein eosin staining.Results At week 1,the new bone ratio (BV/TV) was 5.47％ ± 1.90％ in group A and 8.49％ ± 1.26％ in group B,showing no significant difference (P ＞ 0.05);at weeks 4 & 8,the BV/TV were 17.54％ ±2.04％ and 39.73％ ± 4.01％ in group A,significantly lower than those in group B (25.32％ ± 2.15％ and 53.22％ ± 2.94％) (P ＜ 0.05).At weeks 1 & 4,the scaffold degradation ratios (RSV/SV) were 97.33％ ± 2.52％ and 80.60％ ±4.00％,showing no significant differences from those in group B (95.67％ ±3.51％ and 75.22％ ±6.20％) (P ＞ 0.05).At week 8,the scaffold degradation ratio in group A (65.46％ ±4.51％) was significantly higher than that in group B (50.19％ ±4.91％) (P ＜ 0.05).At week 8,hematein eosin staining showed better integration of scaffolds with the femur,faster degradation of the interior scaffolds and greater osteogenetic activity in group B.Conclusion Prevascularization of tissue-engineered bone graft may increase new bone volume and scaffold degradation rate,promoting repair of femoral bone defects in rats.

Objective:To study the physical activity level and its influence factors among residents in one suburb of Beijing,so as to provide specific interventions for different people in different circum-stances and to provide reference for health relevant policy-making in the future.Methods:In the study, 7 31 9 subjects aged 1 8 years or above were involved.The self-designed questionnaires based on Health Belief Model (HBM)had acceptable validity and reliability.The physical activity levels were calculated to classify sufficient or insufficient amount by a thousand-step equivalent greater than or equal to 6 or 1 0. Multiple variable Logistic regression was used to explore the influence factors of the physical activity among the residents.Results:The residents’median amount of physical activity in the suburb district of Beijing were 9.1 thousand-step equivalent with quartile of (3.8,20.4).The percentages of the thou-sand-step equivalent greater than or equal to 6 or 1 0 were 63.7% and 47.7%,respectively.The median amounts of physical activity from work or household chores,transportation and recreation physical activi-ties were 4.0,1 .0,0.0 and the components of the total amount of physical activity from those were 61 .7%,1 8.3% and 20.1 %,respectively.There were 8.6% residents whose life did notinvolve moder-ate or vigorous intensity activities.By using factor analysis,five factors were extracted from the scale based on the HBM;These factors together contributed to 63.7% of the sum of the squared loadings.The differences of physical activity levels on education level,age,gender,self-efficacy,cues,subjective and objective barriers were statistically significant (P <0.05).Those who were female,with older age,lower education level,higher self-efficacy,fewer cues,fewer subjective and objective barriers preferred to do more physical activities.Conclusion:The physical activity levels among the residents in the suburb dis-trict of Beijing are moderate and high,and most amount of physical activities from work or household chores.Those who are male and whose ages are from 1 8 to 29 years and whose education levels are of university or above should be focused on intervention.Specific interventions should be developed for dif-ferent people in different situations;More attention should be paid to improve the residents’self-efficacy and reduce the subjective and objective barriers of physical activity,and we also should actively advocate people to have more leisure exercise so as to improve the physical activity level among all residents.

This study selected the top five general hospitals in the United States of America, collected the practice information of acupuncturists through the official website, and analyzed the professional department, expertise, professional title, education background, residency, fellowship, board certification, etc. of these acupuncturists to understand the practice situation of acupuncturists in the hospitals. The results of the study showed the practice of acupuncturists in the United States of America is improving. With further localization, locally trained acupuncturists have gradually become the main body of acupuncturists; acupuncture treatment is still mainly for analgesia, but the scope of treatment continues to expand, and departments that accept acupuncturists are gradually increasing. However, the group of licensed acupuncturists is still a minority in the group of acupuncturists in the United States of America. Native American physicians still use acupuncture as an alternative option and neglect to obtain a license. The entry barrier for acupuncturists still needs to be raised.

Objective To investigate the impacts of midazolam on the proliferation,migration and invasion of breast cancer MDA-MD-231 cells and its regulation on cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)/cAMP response element binding protein(CREB)signaling pathway.Methods MDA-MD-231 cells were cultured in vitro and grouped into control group,midazolam L group,midazolam M group,midazolam H group,and midazolam H+Sp-cAMPS group.Midazolam L group,midazolam M group,midazolam H group were treated with 5μmol/L,10μmol/L,and 20μmol/L of midazolam,respectively,midazolam H+Sp-cAMPS group was added with 20μmol/L of midazolam+10μmol/L of Sp-cAMPS 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide(MTT)assay was applied to detect cell proliferation;The migration ability was detected by cell scratch test;Transwell test was applied to determine the invasion ability of cells;Cell apoptosis was detected by flow cytometry;The expression level of cAMP was detected by enzyme-linked immunosorbent assay(ELISA);Western blot was applied to verify the expression of phosphorylated(p-)PKA/PKA,p-CREB/CREB protein.Results Compared with control group,the cells in midazolam L group,midazolam M group,midazolam H group showed apoptosis,the apoptosis rate was obviously increased,the cell proliferation,migration and invasion abilities and the expression levels of cAMP,p-PKA/PKA,p-CREB/CREB proteins were obviously decreased(P<0.05);Compared with midazolam H group,midazolam H+Sp-cAMPS group had good cell growth,obviously reduced apoptosis rate,and obviously increased cell proliferation,migration and invasion abilities,and the expression levels of cAMP,p-PKA/PKA,p-CREB/CREB proteins(P<0.05).Conclusion Midazolam may inhibit the proliferation,migration and invasion of breast cancer cells by inhibiting the activation of cAMP/PKA/CREB signaling pathway.

Objective To systematically evaluate the biomechanical recovery of drilled holes in the femur in SD rats.Methods Eighteen female SD rats were randomized into 3 even groups (n =6).Models of 2-mm drilled holes in bilateral femurs were established in groups A and B with 2 holes on each side while no drilling was performed in group C.Samples were harvested in group A at postoperative 4 weeks,in group B at postoperative 8 weeks while at both 4 and 8 weeks in group C.The samples were evaluated in terms of linear elasticity (compression test),viscoelasticity (relaxation and creep tests) and durability (fatigue failure test).Micro-CT scan was performed to measure the bone volume fraction (BV/TV) and bone mineral density (BMD) of new bone.Sirus red staining was performed to measure regeneration of type Ⅰ collagen of new bone.Results The elasticity modulus,maximum load,compression strength and conditional yield limit in groups A were significantly lower than those in group B which were also significantly lower than those in group C (P ＜ 0.05).At 7,200 s,the relaxation (14.56 ±0.69 MPa) and creep variation (11.37％ ± 0.70％) in group A were significantly higher than those in group B (11.06 0.63 MPa and 8.98％ ± 0.40％) which were also significantly higher than those in group C (6.99 ±0.56 MPa and 5.10％ ±0.23％) (P ＜ 0.05).At the constant amplitude loads from 20 N to 200 N,from 20 N to 300 N and from 20 N to 400 N,the recycling numbers in group A (6,044.3 ±879.7,4,093.3 ±628.5 and 1,919.3 ±847.5) were significantly lower than those in group B (10,192.3 ± 1,109.1,6,750.6 ± 818.0 and 3,376.6 ± 671.3) which were also significantly lower than those in group C (28,068.3 ±2,702.6,11,788.3 ± 1,141.6 and 5,296.3 ± 735.0) (P ＜ 0.05).By micro-CT scan,the BVT and BMD in group A were significantly lower than those in group B which were also significantly lower than those in group C (P ＜ 0.05).The sirus red staining showed the type Ⅰ collagen in the bone defect area was completely regenerated in group B.Conclusion Systematic biomechanical measurements may actually detect the characteristics of biomechanical recovery of bone holes in SD rats,enriching the basic research on the bone damage repairing progress.

As an active hydroxyanthraquinone ingredient, emodin is abundant in Chinese medicine herbs, such as Rheum palmatum, Polygonum cuspidatum and Polygonum multiflorum.Modern pharmacological studies have shown that emodin has a variety of pharmacological activities including anti-tumor, anti-inflammatory and immunoregulatory, antibacterial and anti-viral effects, myocardial protection, neuroprotection, renal protection, bone protection, antifibrosis and so on, which indicate its high medicinal value and broad application prospects.This article aims to summarize the progress in the pharmacological activity and mechanism of action of emodin published in domestic and international journals over the last 5 years and highlight the potential targets and molecular signaling pathways linked with emodin, so as to provide some clues and references for further development and clinical application of emodin.

ObjectiveTo explore the mechanism and pathway of Gandou Fumu decoction （GDFMD） in the development of liver fibrosis in Wilson's disease （WD）. MethodFirst， 30 TX-j mice were randomly divided into the model group， high-dose， medium-dose， and low-dose GDFMD groups， and penicillamine group， with six mice in each group， and another six wild-type mice were used as the normal group. The high-dose， medium-dose， and low-dose GDFMD groups were intragastrically administered drugs of 13.92， 6.96， 3.48 g·kg^-1. In the penicillamine group， 0.1 g·kg^-1 of penicillamine was given by intragastric administration. The model group and the normal group were given equal volume of normal saline， once a day， for four consecutive weeks. Samples were collected four weeks after gavage， and enzyme-linked immunosorbent assay （ELISA） was used to detect type Ⅲ procollagen peptide （PCⅢ）， collagen type Ⅳ （Col Ⅳ）， hyaluronic acid （HA）， and laminin （LN）. Hematoxylin-eosin （HE）， Masson， and picric acid-Sirus red collagen （Sirus Red） staining were used to observe the histopathological changes of liver fibrosis. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， immunohistochemistry， and Western blot were used to observe the expressions of α-smooth muscle actin （α-SMA） and collagen type Ⅰ （Col Ⅰ）， which were related to the activation of hepatic stellate cells （HSCs）. The expression of miR-29b-3p was observed by Real-time PCR. The expression of Unc-51-like kinase 1 （ULK1） and its downstream-related factors were observed by Western blot. The downstream genes of miR-29b-3p were verified by the dual luciferase reporter gene detection method. ResultCompared with the normal group， the four items of liver fibrosis （PCⅢ， Col Ⅳ， HA， and LN） in the model group were significantly abnormal （P<0.01）， and the pathology was significantly abnormal. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ， as well as α-SMA mRNA and Col Ⅰ mRNA was up-regulated （P<0.05， P<0.01）， and miR-29b-3p expression was down-regulated （P<0.01）. ULK1， p-ULK1， autophagy-related gene 13 （Atg13）， p-Atg13， Beclin-1， FAK family kinase-interacting protein of 200 kDa （FIP200）， activating molecule in BECN1-regulated autophagy protein 1 （AMBKA1）， and microtubule-associated protein 1 light chain 3Ⅱ/Ⅰ（LC3Ⅱ/Ⅰ） were up-regulated （P<0.05， P<0.01）. p62 protein expression was down-regulated （P<0.01）. Compared with the model group， the four items of liver fibrosis in the high-dose， medium-dose， and low-dose GDFMD groups and the penicillamine group were significantly improve （P<0.01）， and the pathological conditions were improved. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ， as well as α-SMA mRNA and Col Ⅰ mRNA was down-regulated （P<0.05， P<0.01）， and the expression of miR-29b-3p was up-regulated （P<0.01）. ULK1， p-ULK1， Atg13， p-Atg13， Beclin-1， FIP200， AMBKA1， and LC3Ⅱ/Ⅰ were down-regulated （P<0.05， P<0.01）， and p62 protein expression was up-regulated （P<0.01）. The prediction software predicted that there was a binding site between miR-29b-3p and ULK1. The dual-luciferase reporter gene detection method indicated that the luciferase activity of the ULK1-WT plasmid-transfected cell group was reduced when miR-29b-3p mimics were co-cultured （P<0.01）. ConclusionGDFMD can regulate ULK1-mediated autophagy by up-regulating miR-29b-3p and further exert its anti-hepatic fibrosis effect in Wilson's disease.