Objective To compare four methods of monitoring vascularization of tissue engineered bone in the rhesus so as to find our the best. Methods Twenty-five lower limbs of 13 rhesuses were used in this study to make models of tibial diaphyseal defect of 20mm which were to be fixed with an AO reconstruction plate of 7 holes. The monkeys were randomly divided into five groups according to defect filling materials: group A:?-tricalcium phosphate (?-TCP) and bone marrow stromal cells (BMSCs) and blood vessel bundles; group B:?-TCP and blood vessel bundles; group C:?-TCP and BMSCs; group D:?-TCP; group E: blank. Perfusion weighted MR imaging (PWMR), X-ray, radionuclide imaging and histological examinations were carried out at weeks 4, 8, 12 postop- eratively. The maximum slope rates of the single intensity-time curve (SS_(max)) and values of baseline (Sl_(?))were calculated at the same time points. Transmittances of the X-ray films were assessed. Ratios between isotope counts in region of interest (ROI) were calculated. Chinese ink perfusion and calculation of blood vessel areas were done for histological examinations, Results Compared with other groups, the SS_(max) in group A was the highest at weeks 4, 8, 12 postoperatively. In group A, the SS_(max) at week eight was significantly higher than that at week four (P= 0. 003), and the SS_(max) and transmittance of X-ray were negatively related at week 12 after operation (rs=-0. 892, P=0. 042), but the SS_(max) and blood vessel area were positively related (rs=0. 894, P=0.041)Conclusions PWMR can be a sensitive, quantitative, noninvasive and non-radiant method to monitor vascularization of tissue engineered bone, because SS_(max) of the single intensity-time curve of PWMR can reflect the most accurately the process of vascularization of tissue engineered bone.

Objective To analyze the manifestation,reason,the processing method of the steel wire implantation with the sereotactic mammography to improve the accuracy of the preoperative positioning.Methods Seventy-nine cases which got the stereotactic steel wire implantation.In 96 lesions, 13 had steel wire displacement.Among them,5 cases got steel wire displacement during the sereotactic process,5 cases got steel wire displacement after the stereotactic process,2 cases got steel wire displacement during the operation,one case did not show the calcification on the postoperative radiography.Results The steel wire displacement occurred in 5 cases during the stereotactic process came from the patients and doctors respectively and the repositioning was needed.The steel wire displacement after the stereoscopic positioning was attributed to the overdose injection of local anesthesia,which led to the mismatch between the depth of Z axis of the mammary gland and the actual depth the computer given,the incorrect method for needle placement,and,neglecting whether the steel wire have got the lesion anchored when pulling out the needle set of steel wire hood,besides,these three kinds of instances above were all exaggerated by the accordion effect.For the displacement within 2 cm,the lesion can be excised toward the pathological change direction according to the position that steel wire prompted and re-place the second steel wire,putting the J-shaped steel wire into the needle hood and taking it out of the body.After repositioning,2 cases had the steel wire prolapse during operation,which resulted from the over-lifting of the steel wire.After placing the steel wire, the radiologist should give an accurate description on the depth and direction to the surgeon and the notch should be taken for incision from the steel wire head end which is proximate to skin.The postoperative specimen from one case had no calcification,which might be related to the condition that the calcification was located in the gland body,which got destruction from the surgical electrical electrotome.The excisionscope should be extended and the short term reexamination is recommended to make sure the complete excision of the calcification.Conclusion It is the gold standard method that implanting the steel wire with the stereotactic mammography to guide the surgical dissecting technique to diagnose non-palpable breast lesion(NPBL).Thorough understanding of the displacement manifestation of implanting steel wire with stereotactic technique and the treatment methods will be helpful in the surgical dissecting guidance.

Objective To observe the effect of low arsenic sub-chronic exposure on blood routine test index in rabbits to pave a way for screening early injury of the low arsenic exposure. Methods Twelve healthy male rabbits were randomly divided into four groups. They were administrated with As at the concentration of 0(control), 10, 50 and 250 μg/L in the drinking water. Blood samples were collected from the vein of the ear edge in 8 weeks, and blood test routine including leukocyte (WBC), lymphocyte (LYM), lymphocyte percentage (LYM%), neutrophil (GRA), neutrophil percentage (GRA%), monocyte (MON), monocyte percentage (MON%), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration(MCHC), RDW, platelets(PLT), mean platelet volume(MPV), platelet hematocrit(PCT) and platelet distribution width (PDW), were detected by the ABX-60 hemocyte analyzer. Results Compared with the control group, the WBC, GRA and GRA% increased in 0, 50 and 250 μg/L groups, but there was no significance(P > 0.05). PLT and MPV had a statistical significance in 4 groups(F = 4.07,4.20, all P < 0.05). Compared with the control group[(292.00±16.97)×109/L, (7.10±0.99)fL], PLT decreased in the 250 μg/L group [(221.33±22.50)×109/L] and MPV decreased in the 50μg/L group [(5.57±0.46)fL] significantly (P < 0.05). The other index didn't change obviously. Conclusions Sub-chronic low level arsenic exposure may lead to the change in the blood system. The blood routine test may be considered for early injury of the arsenic poisoning.

OBJECTIVETo evaluate the different effect on the expression of Calcitonin gene related peptide (CGRP)and neuropeptide Y (NPY) between tissue engineered bone with vascular bundle graft in vivo and that with sensory nerve tract graft in vivo.

METHODThirty-six healthy New Zealand rabbits were divided into 3 groups randomly and equally: vascular bundle group (A), sensory nerve tract group (B), tissue-engineering group (C). Group A segmental bone defect of 1.5 cm long was made at the right femur in each animal. After plate fixation, the defects were implanted respectively with the engineered bone prepared in the above-mentioned 3 methods. At 3, 6 and 12 months post-operatively, the distribution of CGRP and NPY in the new bone were detected by immunohistochemistry and analyzed semi-quantitatively by image analysis software.

RESULTSCGRP and NPY immuno-histochemical results indicated their contents increased significantly in all 3 groups as time passed (P = 0.000). Compared with group B, the contents of CGRP and NPY in group A significantly increased at 3 months (P = 0.000), but there was no statistic difference between them at 6 or 12 months (P > 0.05). The expression of CGRP and NPY in both group A and B were significantly more than that in group C at 3, 6 or 12 months (P = 0.000).

CONCLUSIONImplantation of vascular bundle into tissue-engineered bone can significantly improve the CGRP and NPY contents at early 3 months comparing with Implantation of sensory tract into tissue-engineered bone, but the changes are not significant at 6 or 12 months post-operatively.

Animals ; Blood Vessels ; transplantation ; Bone Substitutes ; Calcitonin Gene-Related Peptide ; metabolism ; Disease Models, Animal ; Femur ; injuries ; Male ; Neuropeptide Y ; metabolism ; Peripheral Nerves ; transplantation ; Rabbits ; Random Allocation ; Tissue Engineering

OBJECTIVETo observe the normal structure of lumbar plexus in the virtual Chinese Human (VCH) Female I and Male III and establish a digitized visible model of their lumbar plexus.

METHODSThe cross-sectional images from the VCH Female I and Male III dataset were reviewed to study lumbar plexus structures on a section-by-section basis. The nerve roots, major psoas muscle and blood vessels were also observed. Three-dimensional computerized reconstructions of lumbar plexus and its adjacent structures were conducted from these data using Amira 3.1 (TGS) imaging software respectively.

RESULTSThe three-dimensional reconstructed visible models perfectly displayed the anatomic relationships of lumbar plexus structures and their adjacent structures.

CONCLUSIONSVCH Female I and Male III dataset can provide complete and accurate data of main structure of lumbar plexus. The digitized models of lumbar plexus offer unique insights into the complex anatomy, and morphologic data for imaging diagnosis and treatment of the injury of lumbar plexus.

Anatomy, Cross-Sectional ; Female ; Humans ; Imaging, Three-Dimensional ; Lumbosacral Plexus ; anatomy & histology ; Male ; Models, Anatomic ; Visible Human Projects

BACKGROUND:Tissue factor (TF) is the initiation factor of the extrinsic coagulation pathway, and plays a critical role in the process of thrombosis. This study aimed to investigate the expression of TF and to explore their clinical effect on the pulmonary artery after acute pulmonary thromboembolism. METHODS:Thirty-four Japanese white rabbits (Level II animals) supplied by Tianjin Medical University were randomly assigned into:group A, specimens of the pulmonary artery taken 3 hours after pulmonary embolism (n=8); group B, specimens of the pulmonary artery taken 8 hours after pulmonary embolism (n=8); group C, specimens of the pulmonary artery taken 24 hours after pulmonary embolism (n=8); and control group, pseudo-operations performed without injection of autologous blood clots (n=10). The animal model of pulmonary thrombo-embolism was established by injection of autologous blood clots into the jugular vein through a 5F catheter, and was confirmed by digital subtraction angiography. The mRNA expression of TF in different parts of the pulmonary artery was accessed by RT-PCR. Theq test was used if there was a significant difference in a given continuous variable among the three groups assessed by ANOVA. The experiment equipment was supplied by the State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, the Chinese Academy of Medical Sciences and Peking Union Medical College. RESULTS:The TF expression in the specimen adjacent to emboli was stable at 3, 8 or 24 hours after embolism. The mRNA expression of TF at 3 and 8 hours after embolism was lower in the specimens taken from the distal end of the morbid pulmonary artery than those adjacent to emboli. While at 24 hours after embolism, there were similar mRNA levels in specimens either adjacent or distal to emboli. CONCLUSION:The high level of TF expression in pulmonary artery tissue adjacent to emboli could lead to locally increased coagulation activity, indicating the necessity of initiating anti-coagulation treatment as soon as possible after acute pulmonary embolism.

OBJECTIVETo investigate the correlation between trauma and pulmonary thromboembolism.

METHODSComminuted fractures and extensive soft-tissue contusion at both hind limbs were made by a falling weight from a height in 16 rabbits. Lung perfusion scanning was performed to obtain the radioactivity counts before trauma, at 1 h, 48 h and 96 h after trauma. All the data were divided into 4 groups based on the above 4 time points. The rabbits were sacrificed when positive findings on the pulmonary perfusion scanning appeared. Their lungs were harvested to be paraffin-embedded and stained with hematoxylin-erosin method for histological examination of thromboembolism. The randomized block design ANOVA and the method of least significant difference (LSD) were used for statistical analysis of the radioactivity counts.

RESULTSThe histological findings showed that pulmonary embolism developed in 6 of the 16 rabbits (37.5%). Five of the 6 pulmonary embolism rabbits presented neither clinical symptoms nor positive pulmonary embolism manifestations in the lung perfusion scanning. A significant difference was found in lung perfusion radioactivity between the pre-traumatic, post-traumatic 1h groups and post-traumatic 48 h and 96 h groups(P less than 0.05).

CONCLUSIONSFractures of the hind limbs accompanied with extensive soft-tissue contusion may cause pulmonary micro-embolism that is not sensitive to lung perfusion scanning and tends to have no clinical symptoms. Pulmonary embolism development may take more than two days after trauma.

Animals ; Female ; Fractures, Bone ; complications ; Male ; Pulmonary Embolism ; etiology ; Rabbits ; Wounds and Injuries ; complications

Objective To explore the clinical value of indocyanine green (ICG) fluorescence navigation combined with carbon nanoparticles (CNP) in sentinel lymph node biopsy (SLNB) for patients with early breast cancer. Methods A total of 294 early breast cancer patients with axillary node negative in Department of Breast Surgery, Foshan Hospital Affiliated to Sun Yat-Sen University from June 2013 to April 2016 were retrospectively analyzed. Of the patients, 149 cases underwent SLNB with ICG combined with CNP (combination group), while 145 cases underwent SLNB with methylene blue alone (MB group). If the intraoperative pathology results of sentinel lymph nodes (SLNs) were negative, axillary lymph node dissection (ALND) was avoided. The SLNs detection rate, detection number, metastatic SLNs detection rate in SLNB were compared between two groups. The influence of age and body mass index (BMI) on SLNs detection rate was also analyzed. Results In the combination group, subcutaneous lymphatic channels were successfully visualized in 145 patients, and the detection rate was 97.3％(145/149). The fluorescence of SLNs was successfully detected in 143 patients, and the detection rate was 95.9％(143/149). The detection rate of SLNs was higher in the combination group than that of methylene dye alone group (97.9％vs. 91.0％,χ2=6.902,P<0.05). The average number of detected SLNs was higher in the combination group than that of methylene dye alone group (4.5±1.6 vs. 3.2±1.5,t=4.476,P<0.05). Fifty-eight metastatic SLNs were found in 715 SLNs in the combination group (8.1％), and 26 in 544 SLNs in MB group (4.7％). The detection rate was significantly higher in the combination group than that of methylene dye alone group (χ2=13.714,P<0.01). Age and BMI showed no influence on the detection rate and accuracy of SLNB in two groups (P>0.05). Conclusion The combined tracing of ICG fluorescence and carbon nanoparticles for SLNB has showed a better stability and operability in patients with early breast cancer, which is recommended to be a new SLNB method.

OBJECTIVETo explore the isolation, in vitro culture and chondrogenic differentiation of goat bone marrow mesenchymal stem cells (BMSCs).

METHODSBone marrow was harvested from a 10-month-old Chinese goat for adherent culture of the BMSCs in vitro. Flow cytometry was performed to detect the cell surface markers of the BMSCs of the fourth generation. The induction medium (containing 10% fetal bovine serum, high-glucose DMEM, 6.25 microg/ml insulin, 6.25 microg/ml transferrin, 50 microg/ml vitamin C, 100 nmol/L DXM and 10 ng/ml transforming growth factor-beta1) was then applied for chondrogenic differentiation. Cytochemical staining, RT-PCR and Western blotting were performed to detect the expressions of type II collagen and aggrecan in the cells at the time points of 0, 1, 2 and 4 weeks.

RESULTSThe goat BMSCs grow well in vitro with a high purity in the fourth generation. The expression of chondrocyte phenotypes were observed at 1, 2 and 4 weeks, which became more obvious as the culture prolonged. The mRNA and protein expression of type II collagen and aggrecan in the BMSCs increased obvious after the induction and had reached a satisfactory level by 2 weeks.

CONCLUSIONGoat BMSCs have the potential to differentiate into chondrocytes in vitro, and the results of this study provide the experimental basis for application of goat BMSCs in bone and cartilage tissue engineering in vivo.

Aggrecans ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Chondrocytes ; cytology ; Collagen Type II ; genetics ; metabolism ; Goats ; Mesenchymal Stromal Cells ; cytology ; RNA, Messenger ; genetics ; metabolism

OBJECTIVETo observe the osteoblasts transfected with green fluorescent protein (GFP)by adenovirus vector expressed in vitro and traced it in vivo in order to research the feasibility of GFP as a tracer of seeding cells for tissue engineering.

METHODSCFP were transfected into the osteoblasts which derived from adult human bone marrow stromal cell (hBMSc) by adenovirus vector after being packed in 293A cells. The nontransfected hBMSc was used as the control group. The osteoblasts in each group were observed under an inverted phase contrast microscope and fluorescence microscope. The expressive efficiency of GFP was examined by flow cytometry,and alkaline phosphatase (ALP) activities and osteocalcin (OCN) synthesis. After eight days of the transfection,the osteoblasts were implanted into the muscle of nude mouse thigh while the non-transfected osteoblasts were also implanted as a control. Four and eight weeks after the operation, the nude rats were killed and the continuous tissue sections were examined using fluorescence microscopy after adjacent sections were performed by immunohistochemistry or routine HE staining.

RESULTSThe green fluorescence was shown the transfected osteoblasts which derived from bone marrow. The rate of positive expression was over 75%. After eight days of the transfection, the marker proteins of the surface of the osteoblasts showed extremely efficient expression of CD29 and CD44, but the CD34 expressed negative. Either ALP or OCN of the osteoblasts was no significant difference between the two groups (P > 0.05) four and eight days after the transfection. The GFP were obviously expressed in nude mouse both at four and eight weeks, meanwhile it did not harm on the morphology and function of the transfected osteoblasts whose immunohistochemistry examination showed positive reaction.

CONCLUSIONSGFP could be transfected osteoblasts effectively in vitro and traced in vivo in nude mouse. It may be an optimal tracer for living cells on tissue engineering research.

Adult ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Green Fluorescent Proteins ; genetics ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Osteoblasts ; cytology ; Tissue Engineering ; methods ; Transfection