OBJECTIVETo observe the role of green fluorescent protein (GFP) in tracing rhesus bone marrow stromal cells (rBMSCs) during tissue-engineered bone formation in vivo.

METHODSAd5.CMV-GFP was amplified by infecting QBI-293A cells, and the bone marrow was harvested from the ilium of adult male rhesus to obtain rBMSCs, which were cultured and passaged in vitro. GFP was transfected into the third-passage rBMSCs via adenovirus vector and the labeled cells were inoculated into absorbable HA scaffold and cultured for 3 days, with untransfected rBMSCs as control, before the cell-matrix compounds were implanted into the latissimus dorsi muscles of rhesus. Samples were harvested at 6 week and embedded in paraform, and ground sections of the bone tissue were prepared to observe green fluorescence under laser scanning confocal microscope. Propidium iodide staining of the sections was also performed for observation.

RESULTSThe rBMSCs grew well after GFP transfection, and green fluorescence could be seen 24 h after the transfection and became stronger till 48 h, with a positive transfection rate beyond 80%. Six weeks after cell implantation, the rBMSCs labeled by GFP-emitted green fluorescence were detected in the bone tissue under laser scanning confocal microscope.

CONCLUSIONGFP can effectively trace BMSCs during bone tissue engineering, and the transplanted BMSCs constitute the main source of bone-forming cells in bone tissue engineering.

Animals ; Bone Substitutes ; Cell Differentiation ; Cells, Cultured ; Green Fluorescent Proteins ; genetics ; metabolism ; Macaca mulatta ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Microscopy, Confocal ; Tissue Engineering ; methods ; Transfection

OBJECTIVETo observe the distribution of the nerve fibers in the bone tissue and the entry points of these fibers into the bone.

METHODSThe adult tibia was used for the ground sections which were afterwards made into the slice sections by decalcification in ethylenediamine tetraacetic acid (EDTA). The ground sections were stained in silver and the slice sections were stained in silver and haematoxylin and eosin (HE) respectively. Then, the samples of the transmission electron microscope and the atomic force microscope were made and observed.

RESULTSIn the human long bone tissue, many nerve fibers were distributed in the membrane, cortical bone, cancellous bone and marrow. The nerve fibers entered the bone from the nutrient foramen, and passed through the nutrient canal, Haversian's canal and Volkmann's canal, and finally into the bone marrow. In the nutrient canal, the nerve fibers, mainly the medullary nerve fibers, followed the blood vessel into the bone. In the cortical bone, the nerve fibers also followed the blood vessels and were mainly distributed along Haversian's canal and Volkmann's canal. In the bone trabecular and bone marrow, there were many nerve fiber endings arranged around the blood vessels, mainly around the tunica media of medium-size arteries in the marrow and around capillary blood vessels, and a few scattered in the bone marrow. There were sporadic nerve endings in epiphyseal plate and no nerve fibers permeated epiphysis to diaphysis. No distribution of nerve fibers could be found in cartilaginous part.

CONCLUSIONSThere are many nerve fibers in bone and the nerve passageway is nutrient foramen, Volkman's canal, Haversian's canal and bone marrow.

Adult ; Humans ; Microscopy, Atomic Force ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Nerve Fibers ; ultrastructure ; Staining and Labeling ; Tibia ; anatomy & histology ; innervation ; ultrastructure

OBJECTIVETo discuss the experience with three-dimensional reconstruction technique in initial clinical application in gastrocnemius muscle flap surgery.

METHODFrom 2007 to 2008, 7 patients received gastrocnemius muscle flap surgeries to repair the wounds. Preoperative CT angiography or magnetic resonance imaging (MRI) was performed after injection of the contrast media for individualized three-dimensional gastrocnemius muscle flap reconstruction using Amira4.1 software. According to the size of the defect in the wound, individualized three-dimensional gastrocnemius muscle flap was designed and harvested from the posterior leg.

RESULTSIndividualized three-dimensional reconstruction of the gastrocnemius flap was performed in 7 cases, and the reconstructed flaps clearly displayed the blood vessels, skin and the adjacent three-dimensional structures. In 6 cases the main perforating branched and trunk of the blood vessels in the designed flap were consistent with the surgical findings; in 1 case, the perforating branches failed to be clearly displayed in the designed flap, and surgical examination identified perforating branches with an average diameter of 0.5 mm (minimally 0.3 mm). The flaps survived in all the 7 cases.

CONCLUSIONSThree-dimensional reconstruction of the gastrocnemius flap based on the lower limb CT angiography or MRI allows three-dimensional observation of the anatomy of the flap and accurate marking of the extent of the flap to be harvested, therefore avoiding intraoperative injuries to the blood vessels to better survival of the flaps.

Humans ; Imaging, Three-Dimensional ; methods ; Magnetic Resonance Imaging ; Muscle, Skeletal ; diagnostic imaging ; surgery ; Preoperative Period ; Surgical Flaps ; Tomography, X-Ray Computed

OBJECTIVETo explore the influence of obesity on surgical procedure and short-term surgical outcome in patients with gastric carcinoma.

METHODSA total of 426 patients with gastric carcinoma underwent laparotomy in our hospital during January 2006 and June 2008. All the patients were divided into obesity group and non-obesity group according to body mass index (BMI). The thickness of subcutaneous fat (SCF), abdominal anterior-posterior diameter (APD) and transverse diameter (TD) at the umbilicus level were measured by abdominal CT. Furthermore, the surgical data and postoperative conditions including short-term outcome were reviewed and compared between two groups.

RESULTSThe incidence of obesity was 29.8% in gastric carcinoma patients. Mean values of SCF thickness, APD and TD in obesity group and non-obesity group were (21.8+/-7.1) mm vs (14.4+/-7.5) mm, (223.2+/-24.6) mm vs (181.8+/-23.5) mm and (323.6+/-23.8) mm vs (285.8+/-24.4) mm (P=0.000). Longer operative time (P=0.007) and less amount of dissected lymph nodes were found in obesity group as compared to non-obesity group (P=0.000). Also, obesity group lasted a longer postoperative period of fever (P=0.000) and experienced more post-operative complications (P=0.005) than non-obesity group did.

CONCLUSIONSAbdominal CT scan may display the abdominal shape of gastric carcinoma patients, hence, it is useful to evaluate the difficulty of surgical procedure. These patients may involve in complicated surgical procedure and worse short-term outcome due to obese abdominal shape. Therefore, perioperative management should be emphasized for these patients.

Abdomen ; surgery ; Adult ; Aged ; Aged, 80 and over ; Body Mass Index ; Female ; Gastroplasty ; Humans ; Male ; Middle Aged ; Obesity ; Stomach Neoplasms ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo investigate the epidemiological characteristus of human caliciviruses (HuCVs) among children under 5 years of age with acute diarrhea and to estimate the disease burden in Lulong county.

METHODSHuCVs were detected by enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). Some PCR amplicons were cloned and sequenced. Phylogenetic tree was constructed for strain characterization. The rate of HuCVs-attributed hospitalization was estimated according to the positive rate of HuCVs detection in fecal specimens collected from hospitalized diarrhea patients.

RESULTSBetween July 1999 and June 2001, 708 fecal specimens were collected, of which 393 rotavirus-negative and 5 rotavirus-positive specimens were detected for HuCVs. Thirty-one point six percentage of fecal specimens from patients with diarrhea was HuCVs positive. Among inpatients, HuCVs positive rate was 17.5%. HuCVs detection was mainly distributed in 3 - 17 mouth-old children, in winter. All 11 strains belonged to NLV GII in which 6 strains GII-3, 2 strains GII-4 and 3 strains GII-7, and they shared 55.1% - 100% nucleotide identity. NLV GII-4 and GII-7 were identified in 2000, while NLV GII-3 and GII-7 in 2001. The preliminary estimate of HuCVs-attributed hospitalization rate was 3.6 per thousand.

CONCLUSIONHuman caliciviruses with different genotypes circulated among children in Lulong county with GII NLVs were the prevalent strains. The disease burden of HuCVs was second to rotavirus.

Acute Disease ; Age Factors ; Caliciviridae ; genetics ; immunology ; Caliciviridae Infections ; complications ; epidemiology ; Child, Preschool ; China ; epidemiology ; Dysentery ; epidemiology ; etiology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Infant ; Inpatients ; statistics & numerical data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Seasons

OBJECTIVETo investigate the protective effect of hepatocyte growth factor (HGF) on protein synthesis in rat cardiomyocytes exposed to gamma-ray irradiation.

METHODSPrimary cultured cardiomyocytes were irradiated with single-dose (20 Gy) gamma ray in the absence or presence of HGF (40 ng/ml) added in the cell culture 3 h before the exposure. Forty-eight hours after irradiation, the total cellular protein was measured and cell cycle analyzed by flow cytometry. The cardiomyoctes were also infected with AdGFP 48 h after irradiation and the fluorescence intensity of the green fluorescence protein (GFP) in the cells determined by flow cytometry 48 h after infection.

RESULTSThe protein synthesis was decreased significantly in the irradiated cardiomyocytes as compared with the control group (P<0.01), but was remedied significantly by incubation of the cells with HGF before the exposure (P<0.05). Flow cytometry revealed much lower mean fluorescence intensity (MFI) of GFP in irradiated cardiomycytes than in cells without the exposure (P<0.01); The MFI was higher in HGF-treated cardiomyocytes than in cells without HGF treatment following the exposure (P<0.01).

CONCLUSIONGamma ray irradiation inhibits protein synthesis in cardiomyocytes, and HGF may attenuate this effect of gamma ray exposure for cardiomyocyte protection.

Animals ; Animals, Newborn ; Cell Cycle ; drug effects ; radiation effects ; Cells, Cultured ; Flow Cytometry ; Gamma Rays ; Green Fluorescent Proteins ; genetics ; metabolism ; Hepatocyte Growth Factor ; pharmacology ; Microscopy, Fluorescence ; Myocytes, Cardiac ; cytology ; metabolism ; Protein Biosynthesis ; drug effects ; radiation effects ; Rats ; Rats, Wistar

OBJECTIVETo study whether tissue engineered bone can repair the large segment bone defect of large animal or not. To observe what character the fascia flap played during the osteanagenesis and revascularization process of tissue engineered bone.

METHODS9 Chinese goats were made 2 cm left tibia diaphyseal defect. The repairing effect of the defects was evaluated by ECT, X-ray and histology. 27 goats were divided into three groups: group of CHAP, the defect was filled with coral hydroxyapatite (CHAP); group of tissue engineered bone, the defect was filled with CHAP + bone marrow stroma cells (BMSc); group of fascia flap, the defect was filled with CHAP + BMSc + fascia flap. After finished culturing and inducing the BMSc, CHAP of group of tissue engineered bone and of fascia flap was combined with it. Making fascia flap, different materials as described above were then implanted separately into the defects. Radionuclide bone imaging was used to monitor the revascularization of the implants at 2, 4, 8 weeks after operation. X-ray examination, optical density index of X-ray film, V-G staining of tissue slice of the implants were used at 4, 8, 12 weeks after operation, and the biomechanical character of the specimens were tested at 12 weeks post operation.

RESULTSIn the first study, the defect showed no bone regeneration phenomenon. 2 cm tibia defect was an ideal animal model. In the second study, group of CHAP manifested a little trace of bone regeneration, as to group of tissue engineered bone, the defect was almost repaired totally. In group of fascia flap, with the assistance of fascia flap which gave more chance to making implants to get more nutrient, the repair was quite complete.

CONCLUSIONSThe model of 2 cm caprine tibia diaphyseal defect cannot be repaired by goat itself and can satisfy the tissue engineering's demands. Tissue engineered bone had good ability to repair large segment tibia defect of goat. Fascia flap can accelerate the revascularization process of tissue engineered bone. And by this way, it augment the ability of tissue engineered bone to repair the large bone defect of goat.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Bone Regeneration ; physiology ; Bone Substitutes ; Cells, Cultured ; Durapatite ; Fascia ; transplantation ; Goats ; Implants, Experimental ; Neovascularization, Physiologic ; Osteogenesis ; Random Allocation ; Stromal Cells ; cytology ; Tibia ; blood supply ; injuries ; surgery ; Tibial Fractures ; surgery ; Tissue Engineering

OBJECTIVETo investigate the main risk factors associated with intra-abdominal infection(IAI) following gastrectomy in gastric cancer patients.

METHODSCase-control study was used to investigate the clinical data of 1728 gastric carcinoma cases retrospectively by Logistic regressive analysis.

RESULTSUnivariate Logistic regressive analysis showed 16 factors, including age, malnutrition, chronic obstructive pulmonary disease(COPD), diabetes mellitus(DM), heart diseases, prothrombin time, lymphocyte count, tumor size, ascites, invasion to the adjacent organ, neoplasm TNM staging (UICC, 1997), methods of gastrectomy, blood loss, operative time, blood transfusion and extent of lymph nodal dissection,were associated with postoperative intra-abdominal infection. Binary Logistic regression analysis found that extent of lymph nodal dissection(N(2)(+) approximately N(3) and N(2)), invasion to the adjacent organ, DM, operative time, age and lymphocyte count were the independent risk factors associated with mortality.

CONCLUSIONNecessary interventions should be carried out to prevent IAI referring to above risk factors.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Gastrectomy ; adverse effects ; Humans ; Logistic Models ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Postoperative Complications ; etiology ; Risk Factors ; Stomach Neoplasms ; microbiology ; pathology ; surgery ; Surgical Wound Infection ; etiology ; Young Adult

OBJECTIVETo detect the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSc) transferred by retroviral vector carrying human bone morphogenetic protein 7 (hBMP-7) gene.

METHODShBMP-7-expressing replication-deficient retroviral vector(PT-PLNCX2-hBMP7) was reconstructed using clone technique and recombinant DNA technique. BMSc were infected with the virus granules. The protein of BMP-7 gene in transferred cells were determined by immunohistochemistry. The proliferativity of the transferred cell were assayed by methabenzthiazuron (MTT) method and flow cytometer. Alkaline phosphatase (ALP) were also detected using enzyme kinetics.

RESULTSCells transferred by PT-PLNCX2-hBMP7 expressed abundant hBMP7 protein in the cytoplasm. Positive findings were not found in those cells that were not transferred. After infected with virus there were not significant difference of cell proliferation and cell cycle between the cells transferred by hBMP-7 or not (P > 0.05). ALP activity in transferred cells were increased significantly (P < 0.01).

CONCLUSIONShBMP-7 can be transferred and stably expressed in the cultured rabbit bone marrow stem cells. Proliferation and cell cycle of the transferred cell were not affected. hBMP7 gene transfer can be used to induce differentiation of BMSc into osteoblast-like cells.

Alkaline Phosphatase ; biosynthesis ; Animals ; Bone Marrow Cells ; cytology ; enzymology ; Bone Morphogenetic Protein 7 ; Bone Morphogenetic Proteins ; genetics ; Cell Division ; Cells, Cultured ; Gene Transfer Techniques ; Humans ; Osteoblasts ; cytology ; Osteogenesis ; Rabbits ; Stem Cells ; cytology ; enzymology ; Tissue Engineering ; Transforming Growth Factor beta ; genetics

OBJECTIVETo observe the impact of specific short hairpin RNA (shRNA) targeting survivin gene on tumorigenesis and angiogenesis of human brain glioblastoma U251 cells in vivo of nude mice.

METHODSU251 cells, U251-SR cells transfected stably with shRNA eukaryotic expression vector pWH1-SR targeting survivin gene, and U251-P cells transfected stably with blank pWH1 vector, were inoculated respectively into subcutaneous tissue in flank of 15 nude mice (each group 5 mice), and the tumor growth status was observed and measured. Protein expressions of survivin, proliferating cell nuclear antigen (PCNA) and factor VIII related antigen (F VIII RAg) were investigated by immunohistochemistry SABC method, apoptotic cells were screened by TUNEL method, furthermore proliferative index (PI), apoptotic index (AI) and microvessel density (MVD) were measured respectively in each group of tumor specimens.

RESULTSComparing with those in U251 and U251-P groups, in U251-SR group, the tumorigenesis time delayed, tumor grew slowly, both tumor volume and tumor weight decreased significantly (P < 0.01 for both); Survivin protein expression was down-regulated markedly; PI and MVD decreased significantly, whereas AI increased remarkably (P < 0.01 for all).

CONCLUSIONSThe specific shRNA targeting survivin gene can inhibit significantly tumorigenesis and angiogenesis of U251 cells in vivo.

Animals ; Apoptosis ; Brain Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Glioblastoma ; metabolism ; pathology ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Mice ; Mice, Nude ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Transplantation ; Neovascularization, Pathologic ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; Repressor Proteins ; Transfection