Objective To explore the method of primary culture and biological characteristics of aortic vascular smooth mus-cle cells (VSMC)in mice,providing experimental material for cellular and molecular scientific research of vascular disease. Methods Thoracic and abdominal aortas in mice were isolated and VSMC were obtained by using improved method of tissue piece inoculation.Digested with trypsin and passaged,VSMC were purified with differential adherence method.The conditions of cellular morphology and growth were observed under inverted phase contrast microscope,and VSMC were identified with hematoxylin-eosin (HE)staining and immunofluorescence.Results VSMC were isolated successfully and grown vigorously with good bioactivity,the cells had a radial or typicalpeak-valleylike growth,and showed fusiform,abundant cytoplasm with large and round or oval nucle-us by HE staining,the expressions of specific cytoplasmicα-smooth muscle actin were positive by immunofluorescence stain.Conclu-sion It can isolate and cultivate VSMC with high purity and good activity under in vitro conditions with simple,economical,reliable method.

Objective: To research the effect and mechanism of how chitosan restrain fat-absorbtion, and provide references for developing a kind of nutritional and healthy fastfood. Methods: 1.In vitro digestion: Phosphomolybdic acid colorimetry; 2.In vivo experiment:240 rats were divided into four groups and fed with different diets: basal diet(Group A), high fat diet (Group B), chitosan supplemented diet (Group C), high fat and chitosan supplemented diet (Group D). The fat and chitosan content(%) of diet were A(10,0), B(60,0), C(8,20), D(34.3,14.8) respectively. Determine the blood lipids and fat output in dung, and observe the weight gain and swimming time. Results: 1.In vitro experiment showed chitosan acted on cholic acid at the best proportion 1∶8. 2.In vivo experiment showed chitosan can restrain fat-absorbtion, and increase fat output. Swimming in ice-water showed chitosan improved rats performance. Compared with group B, the blood lipids and weight of group C and D had no obvious change, but the fat-output much increased.

Background Chinese dietary reference intakes for calcium are largely based on foreign studies.We undertook metaregression to estimate calcium requirements for Chinese adults derived from calcium balance data in Chinese adults.Methods We searched PubMed,Cochrane CENTRAL,and SinoMed from inception to March 5,2014,by using a structured search strategy.The bibliographies of any relevant papers and journals were also screened for potentially eligible studies.We extracted a standardized data set from studies in Chinese adults that reported calcium balance data.The relationship between calcium intake and output was examined by an individual participant data (IPD) and aggregate data (AD) meta-regression.Results We identified 11 metabolic studies in Chinese adults within 18-60 years of age.One hundred and forty-one IPD (n=35) expressed as mg/d,127 IPD (n=32) expressed as mg·kg body wt-1·d-1,and 44 AD (n=132) expressed as mg/d were collected.The models predicted a neutral calcium balance (defined as calcium output (Y) equal to calcium intake (C)) at intakes of 460 mg/d (Y=0.60C+183.98) and 8.27 mg·kg body wt-1·d-1 (Y=0.60C+3.33)for IPD,or 409 mg/d (Y=0.66C+139.00) for AD.Calcium requirements at upper intakes were higher than that at lower intakes in all these models.Conclusion Calcium requirement for Chinese adults 18-60 years of age approximately ranges between 400 mg/d and 500 mg/d when consuming traditional plant-based Chinese diets.

Objective To differential the biologic characteristics and multiple differentiation potential of mesenchymal stem cells in nucleus pulposus in (NP-MSC) scoliosis patient and patient with degenerative interverthral disc.Methods The human nucleus pulposus-mesenchymal stem cells were isolated and cultured with enzyme digestion from 2 patients of scoliosis and 2 patient with degenerative intervertbral disc separately.Cellular proliferation was detected with MTT assay and trypan blue.The immunophenotype expression of NP-MSC was detected by flow cytometry in scoliosis and degenerative group.The multiple differentiation ability of cells was assessed respectively using alizarin red dye,Oil red O dye and immunohistochemical staining.Results The primary cell morphology of scoliosis NP-MSC was the shape of spindle,while the degenerative NP-MSC was inhomogeneous.However,both of them became spindle shape after passages.The scoliosis NP-MSC was stronger than degenerative one in metabolic activity and proliferation.The percentage of positive antigen expression of CD44,CD105 and CD29 was 97％-100％ in scoliosis NP-MSC group,but 88.7％-97％ in the degenerative group.The expression of mature cell marker CD24 was negative in both groups.Furthermore,the MSC i.isolated from both groups differentiated along the osteogenic,chondrogenic but not adipogenic lineages.Conclusion The scoliosis and degenerative NP contains mesenchymal stem cells.Moreover the scoliosis NP-MSC had stronger ability of cell metabolic activity and proliferation.These cells have multiple differentiation potential with the exception of their adipogenic differentiation ability.

Objective To summarize the curative effect of repairing large area soft tissue defects in heel and crus by flaps with double blood-supply of posterior tibial artery perforators and saphenous nerve nutrient vessels.Methods From January 2006 to February 2012,twenty cases took operation under the guide of Continuous Wave Doppler and design of tibial artery perforator as rotation point.And in all cases,island flaps with the blood supply from saphenous nerve nutrient vessels and tibial artery perforator were retained to repair large area soft tissue defects in heel and crus.In operations,the range of flap area were ranged from 19 cm × 11 cm to 11 cm × 8 cm.Skin flaps incision was up to the patella margin level,low to medial malleolus on edge,former to crus former median line,rear to after crus median line and farthest to the surface of wound on the metatarsophalangeal joint.Results Nineteen cases survived,and 1 case of skin flap mild necrosis at the farthest side took a second-phase line skin flap to repair.Followed-up from 6 months to 24 months was taken in all cases at the mean time of 10 months,with a result of good recovery and no ulceration for the flaps.To varying degree,all flaps recover sense of pain and deep touch.Conclusion There is no wound to posterior main tibial artery in repairing large area soft tissue defects in heel and crus by flaps with double blood-supply from posterior tibial artery perforators and saphenous nerve nutrient vessels,meanwhile to maintain double blood-supply from posterior tibial artery perforators and saphenous nerve nutrient vessels and expand the range of blood supply of posterior tibial artery perforators.In this operation,a blood circulation for the flap can be guaranteed so as for a large wound in heel and crus.

Objective:To analyze the factors related to metastasis of contralateral central lymph node (CLN) in cN0 papillary thyroid car-cinoma (PTC) and discuss the indications for CLN dissection. Methods:We enrolled 149 unilateral PTC patients who underwent total thyroidectomy and prophylactic bilateral (CLN) dissection. This work analyzed the relationship of gender, age, extrathyroidal extension, multifocality, thyroiditis, ipsilateral central lymph nodes, and prelaryngeal lymph node with CLNs. Results:The rates of metastasis to ip-silateral and contralateral central compartments were 73.2%and 23.5%, respectively. In univariate analysis, gender, age, tumor size, multifocality, and thyroiditis were not important in predicting contralateral central compartment lymph node metastasis (P=0.792, 0.097, 0.531, 0.269, and 1.000, respectively);by contrast, extrathyroidal extension (P=0.017), prelaryngeal lymph nodes (P=0.006), and ipsilateral CLNs (P<0.001) are related to CLN metastasis. However, multivariate analysis showed that ipsilateral central metastasis was an independent risk factor for lymph node metastasis in the contralateral central region when the number of ipsilateral central metas-tases is≥3 (P=0.010). Conclusion:Extracapsular invasion, prelaryngeal lymph nodes, and ipsilateral CLN influence the metastases of CLN. Bilateral CLN dissection should be performed when the number of ipsilateral central metastases is≥3 and there is merger of ex-tra-laryngeal lymph nodes or capsule invasion.

Objective To explore the regulation of ROS level and ROS-triggered downstream events on SK-N-MC Ewing sarcoma cells upon apoptasis induction by 2-Methoxyestradiol (2-ME). Methods To detect the reversibility of apoptosis and the alternation of activity of respiratory chain, mitechondria transmembrane potential (△ψm), and cellular ROS level and to explore their association with flow cytometry, clark oxygen electronic node analysis, drug-removal design, and permeability transition (PT) pore stablizing agent. Results SK-N-MC cells were induced to ROS-dependent apoptosis. Apoptosis occured irreversibly after2-ME treatment for 3 h. Upon 2-ME treatment, the activity of respiratory chain was inhibited and the ROS generation was accelerated; the △ψm underwent the increasing within 3h but decreasing after 3h which could be reversed by PT pore stablizing; the ROS level underwent the continuous increasing and PT pore stablizing had no obvious effect on it. Conclusion 2-ME causes the acceleration of ROS generation via inhibiting the activity of respiratory chain and elevating the level of △ψm. ROS plays a signaling role and when total ROS accumulate to a threshold, the PT pore opening and the collapse of △ψm could be induced irreversibly and cell is eventually introduced to death.

AIM: To study the survival, transfer and distribution of bone marrow CD34+/CD45+ cells from transgenic GFP mouse after transplanted into the completed transversional spinal cord rat model. METHODS: The bone marrow cells isolated from transgenic GFP mice were cultured in vitro. The cultured cells were identified by anti-CD34 and anti-CD45 monoclonal antibodies, and were transferred into the end of transection spinal cord. Paraformaldehyde was infused into the left ventricle of the rat model at the 24 h, 48 h, 1 week, 2 weeks, 4 weeks and 8 weeks after cell transplantation. Through sank and frozen, the spinal cord was sectioned at 10 ?m thickness. The green fluorescence positive cells were observed under the fluorescence microscope. CD34+/CD45+ cells were identified by immunohistochemistry staining. RESULTS: Green fluorescence positive cells were found at the head and the end of the completed transection part of spinal cord. Most of the green fluorescence positive cells were distributed in the gray substance of spinal cord. CD34+/CD45+ cells were found by immunohistochemistry staining. CONCLUSION: CD34+/CD45+ cells survived in spinal cord of SD rat, and migrated to the head of the transection part. The distance of migration was extended by the time.

Objective To evaluate the predictive value of serum carbohydrate antigen 50(CA50),tumor specific growth factor(TSGF),and tissue polypeptide antigen(TPA)levels for sensitivity to radiochemotherapy in patients with middle-and advanced-stage breast cancer using a nomogram model.Methods Eighty-two patients with middle-and advanced-stage breast cancer were selected as the study sub-jects.All patients received paclitaxel chemotherapy combined with radiotherapy and were divided into sensitive(n= 57)and insensitive(n= 25)groups according to the Response Evaluation Criteria in Solid Tumors.The general information of the patients,serum expression of CA50,TSGF,and TPA,and their differences before and after treatment were recorded.A nomogram model was constructed,and cali-bration curves,receiver operating characteristic(ROC)curves,and decision curves were used to evaluate the predictive power and clinical utility of the nomogram model.Results Significant differences were observed in tumor diameter,vascular invasion,TNM stage,lymph node metastasis,and degree of differentiation between the two groups(P＜0.05).Compared to those in the sensitive group,the serum expression of CA50,TSGF,and TPA after treatment was higher,and the difference in CA50,TSGF,and TPA was smaller in the insensitive group(P＜0.05).Three predictive variables were identified in the LASSO regression:differences in CA50,TSGF,and TPA.The logistic regression results showed that differences in CA50,TSGF,and TPA influenced sensitivity to radiochemotherapy in middle-and advanced-stage breast cancer(P＜0.05).A nomogram model was constructed using differences in CA50,TSGF,and TPA.Calibration,ROC,and decision curves showed the model's good predictive accuracy and clinical utility.Conclusion Serum expression of CA50,TSGF,and TPA is high in patients with middle-and advanced-stage breast cancer who are insensitive to radiochemotherapy,and differences in CA50,TSGF,and TPA affect their sensitivity to radiochemotherapy.The nomogram model had good predictive value and clinical utility.

Objective To evaluate the efficacy and safety of adjustable single-incision sling(AjustTM) and tension-free vaginal tape(TVT) in the treatment of female stress urinary incontinence(SUI).Methods A total of 88 cases with SUI in our department from January 2011 to December 2015 was retrospectively analyzed,including 62 TVT cases and 26 AjustTM cases.The definite diagnose and grading were based on the medical history,physical examination,cystoscopy,uroflowmetry,provocative test and ICI-Q-SF.All cases were followed up on 6 months after surgery and their uroflowmetry,provocative test and ICI-Q-SF were investigated.Results The average operation time of AjustTM group,(10.0 ± 4.2) min,was shorter than that of TVT group,(50.0± 12.6) min (P＜0.01),and the incidence of operative complications in AjustTM group(3.8％) was less than that in TVT group(6.7％),with significant difference(P＜0.05);The application of these two methods in recent cases showed no significantly statistical difference in objective cure rate,subjective satisfaction,intraoperative blood loss,new-onset OAB,dysuria,perineum discomfort and dyspareunia(P＞0.05).Conclusion AjustTM applied in the treatment of SUI has more advantages than TVT over simplicity,efficiency and safety.