Aging ; physiology ; Animals ; Female ; Male ; Mitochondria, Heart ; enzymology ; Mitochondrial Membrane Transport Proteins ; physiology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Ventricular Function, Left ; physiology

OBJECTIVETo investigate relationship between glucose metabolic disorders and expression of insulin receptor (IR) and tyrosine protein kinase (TPK) in posthepatitic cirrhosis hepatocyte and HBV DNA expression in pancreatic cells.

METHODSTo detect HBV DNA in paraffin-embedded pancreatic and hepatic tissues from 12 posthepatitic cirrhosis patients with positive serum HBV markers by using in situ hybridization (ISH) with a digoxigenin labelled probe. The amount of IR and TPK have been evaluated by immunohistochemical quantitative analysis using image analyzer in hepatocyte of 12 patients positive for HBV markers with posthepatitic cirrhosis in serum. Immunofluorescent histochemical double staining technique was used. HBsAg and IR were observed under confocal laser scanning microscope.

RESULTSEleven of 12 cirrhosis patients? hepatocytes were HBV DNA positive, including 7 patients (7/7) with impaired glucose tolerance (IGT) and 4 patients (4/5) with normal glucose tolerance (NGT). Eight of 12 pancreatic cells were HBV DNA positive, including 7 patients (7/7) with IGT, but only one patient (1/5) with NGT-HBV DNA was found positive in pancreatic cells in significantly more subjects in IGT group than in NGT group (P less than 0.01).IR and TPK amount in hepatocyte of IGT was significantly less than that of NGT patients with posthepatitic cirrhosis (P less than 0.01). IR amount was closely related to the TPK in cirrhosis hepatocyte r=0.82597(P less than 0.01). HBV DNA was mainly localized in the nuclei of hepatocyte and pancreatic acinar and islet cells. Immunofluorescent histochemical double-staining showed that HBsAg was partly localized in the IR positive areas of hepatocytes and pancreatic islet cells.

CONCLUSIONHBV can invade acinar cells of pancreas and islet cells, which might be a direct cause of insulin-dependent diabetes mellitus-like the disorder and insulin absence after HBV infection. Decrease of IR and TPK might be main cause of noninsulin-dependent diabetes mellitus-like disorder after having hepatitis or posthepatitic cirrhosis.

DNA, Viral ; analysis ; Female ; Glucose Metabolism Disorders ; complications ; metabolism ; virology ; Hepatitis B virus ; genetics ; Hepatocytes ; metabolism ; virology ; Humans ; In Situ Hybridization ; Liver Cirrhosis ; complications ; metabolism ; virology ; Male ; Middle Aged ; Pancreas ; cytology ; virology ; Protein-Tyrosine Kinases ; metabolism ; Receptor, Insulin ; metabolism

OBJECTIVETo examine the influence of vascular endothelial growth factors (VEGF) in controlling the growth of an experimental osteosarcoma in mice by performing retrovirus-mediated sFlt-1 gene modification.

METHODSFrom March to October 2010 human osteosarcoma G-292 cells were in vitro infected with retroviral vectors encoding soluble Flt-1 or LacZ gene before transplanted into proximal tibiae of immune deficient SCID mice to establish experimental orthotopic osteosarcoma. Daily observation and biweekly microCT were performed to monitor tumor development and progression till sacrifice at 8 weeks after tumor cell inoculation for histological and molecular analyses.

RESULTSSuccessful transgene expression was confirmed in the culture media of sFlt-1 transduced G-292 cells using ELISA, and with positive X-gal staining of the LacZ transduced cells. Noteworthy tumors were grown in all mice on the tibiae receiving G-292 cell inoculation, with clear detection on microCT images starting 2 weeks after inoculation. Over the time period, tumors derived from sFlt-1 transduced G-292 cells were distinctively smaller in size compared to the ones from wide-type G-292 and G-292-LacZ cells. Histology showed typical osteosarcoma characteristics including severe cellular pleomorphism, bone erosions, and neo-vascularization. Real-time polymerase chain reaction indicated significantly higher sFlt-1 expression in sFlt-1 transduced groups than the wild-type G-292 or LacZ treated groups. Strong expression of oncogenes c-myc and c-fos were also obvious, along with the expression of VEGF in the primary tumor tissue.

CONCLUSIONRetrovirus-mediated sFLT-1 gene modification decelerates the osteosarcoma tumor growth in this murine model.

Animals ; Bone Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Genetic Vectors ; Humans ; Lac Operon ; Mice ; Mice, SCID ; Neovascularization, Pathologic ; metabolism ; pathology ; Osteosarcoma ; genetics ; metabolism ; pathology ; Retroviridae ; genetics ; Transgenes ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism

OBJECTIVESTo examine the gene expression profile of bone morphogenetic protein 2 (BMP-2) and vascular endothelial growth factor (VEGF) during entochondrostosis of mice and explore the expression rules and effects between BMP-2 and VEGF, and to detect the expression of VEGF in BMP-2 induced entochondrostosis in vivo.

METHODScDNA microarray technique with 34,000 genes was used to analyze the gene expression profiles during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of BMP-2 and VEGF was performed with GCOS1.2 software. An experimental model of femoral muscular pouch in 20 mice was adopted. The expression of VEGF was examined by in situ hybridization method and immunohistochemical method in BMP-2 induced entochondrostosis in vivo.

RESULTSThe expression signals of VEGF mRNA and VEGF appeared in cytoplasm during condensation of mesenchymal cell. As the mesenchymal cells differentiated into precartilage, the expression signals decreased in mesenchymal cells, but increased in chondrocytes and kept getting denser in the process of cartilage maturity. The peak expression of VEGF mRNA and VEGF in the experimental group appeared on the 14th day, accompanied by numerous hypertrophic chondrocytes. When mature cartilage calcified and new bone trabecula formed, the expression of VEGF mRNA and VEGF decreased in chondrocytes, but still expressed moderately in the osteoblasts and osteocytes.

CONCLUSIONSThe finding reveals a complex pattern of gene coexpression of BMP-2 and VEGF during the critical period of entochondrostosis. It's feasible for the clinical application of BMP-2 in orthopedics.

Animals ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Cell Differentiation ; genetics ; Chondrocytes ; cytology ; metabolism ; Gene Expression ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Male ; Mice ; Oligonucleotide Array Sequence Analysis ; Osteoblasts ; cytology ; metabolism ; Osteogenesis ; genetics ; RNA, Messenger ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

BACKGROUND: Posterior internal fixation is one of the most common methods for thoracolumbar fractures. There is a lack of systematic evaluation about the efficacy of injured vertebra pedicle screw fixation(IVPSF)versus short-segment pedicle instrumentation (SSPI) for thoracolumbar fracture. OBJECTIVE: To compare the clinical outcomes of IVPSF and SSPI for single thoracolumbar fracture through a METHODS: A computer-based on-line research of PubMed, Medline, Embase, Cochrane Library, CNKI, and WanFang databases was performed for the studies regarding IVPSF versus SSPI for thoracolumbar fracture from 1990 to 2016. meta-analysis. The randomized controlled trials and cohort studies were collected based on the strict criteria of inclusion and exclusion. A meta-analysis was conducted on Revman5.3 sofeware. RESULTS AND CONCLUSION: (1) Eleven articles were enrolled, including 5 English and 6 Chinese ones, involving 689 patients (328 cases for IVPSF and 361 cases for SSPI). (2) The meta-analysis indicated that the operation time, blood loss and mean hospital stay showed no significant differences between two groups. IVPSF showed more effective than SSPI in the kyphotic angle correction and anterior vertebral height recovery at postoperation and 1-5 years of follow-up. Moreover, the incidence of postoperative fixation failure in IVPSF was lower than that in SSPI. (3) These findings suggest that IVPSF that reduces the postoperative fixation failure rate for thoracolumbar fractures provides better kyphosis correction and restoration of anterior vertebral height at post-operation and 1-5 years of follow-up.

OBJECTIVETo investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581.

METHODSGenescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed.

RESULTSD6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005).

CONCLUSIONThese results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30.

Adult ; Aged ; Aged, 80 and over ; Alleles ; China ; F-Box Proteins ; genetics ; Female ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; Pedigree

OBJECTIVETo identify risk factors associated with patients suffered multiple level osteoporosis vertebral compression fractures(OVCFs).

METHODSFrom March 2011 to March 2015, 199 patients suffered osteoporotic were classified into multiple level OVCFs group and single level OVCF group. Multivariate logistic regression analysis were performed to identify risks factors associated with multiple level OVCFs.

RESULTSAll the patients underwent OVCF, including 71 multiple level OVCFs and 128 single level OVCF. There were no differences in the age, gender, BMI, hypertension and diabetes between two groups. While multiple level OVCFs were associated with spinal deformity index SDI[(2<=SDI<4, OR=2.587, 95% CI(1.148, 5.828);SDI>=-4, OR=7.775, 95% CI(3.272, 18.478)], BMD[(T<-4.5SD, OR=2.608, 95% CI(1.038, 6.551)].

CONCLUSIONSSDI and BMD might be the risk factors for multiple level OVCFs.

OBJECTIVETo evaluate the effects and mechanism of radiation-sterilized allogeneic bone sheets in inducing vertebral plate regeneration after laminectomy in sheep.

METHODSTwelve adult male sheep (aged 1.5 years and weighing 27 kg on average) provided by China Institute for Radiation Protection underwent L3-4 and L4-5 laminectomy. Then they were randomly divided into two groups: Group A (n=6) and Group B (n=6). The operated sites of L4-5 in Group A and L3-4 in Group B were covered by "H-shaped" freeze-drying and radiation-sterilized allogeneic bone sheets (the experimental segments), while the operated sites of L3-4 in Group A and L4-5 in Group B were uncovered as the self controls (the control segments). The regeneration process of the vertebral plate and the adhesion degree of the dura were observed at 4, 8, 12, 16, 20 and 24 weeks after operation. X-ray and CT scan were performed in both segments of L3-4 and L4-5 at 4 and 24 weeks after operation.

RESULTSIn the experimental segments, the bone sheets were located in the anatomical site of vertebral plate, and no lumbar spinal stenosis or compression of the dura was observed. The bone sheets were absorbed gradually and fused well with the regenerated vertebral plate. While in the control segments, the regeneration of vertebral plate was not completed yet, the scar was inserted into the spinal canal, compressing the dura and the spinal cord, and the epidural area almost disappeared. Compared with the control segments, the dura adhesion degree in the experimental regenerated segments was much milder (P less than 0.01), the internal volume of the vertebral canal had no obvious change and the shape of the dura sack remained well without obvious compression.

CONCLUSIONSFreeze-drying and radiation-sterilized allogeneic bone sheets are ideal materials for extradural laminoplasty due to their good biocompatibility, biomechanical characteristics and osteogenic ability. They can effectively reduce formation of post-laminectomy scars, prevent recurrence of post-laminectomy spinal stenosis, and induce regeneration of vertebral plates.

Animals ; Bone Transplantation ; methods ; Laminectomy ; methods ; Regeneration ; Sheep ; Spinal Stenosis ; prevention & control ; Spine ; physiology ; Transplantation, Homologous

OBJECTIVETo investigate the diagnostic value of clinical manifestation, laboratory examination and imaging changes for pyogenic spondylitis and to summarize the clinical characteristics of patients with pyogenic spondylitis.

METHODSThe clinical data, of 20 patients with pyogenic spondylitis were diagnosed by histopathological examination from March 2012 to March 2015, were retrospectively analyzed. There were 9 males and 11 females, aged from 43 to 72 years old with an average of 58.9 years. Included 3 cases of cervical vertebrae, 7 cases of thoracic vertebrae, 10 cases of lumbar vertebrae. Patients of blood analysis, erythrocyte sedimentation rate(ESR), C reactive protein(CRP), X rays, CT and MRI were performed before treatment. Visual analogue scale (VAS) was used to evaluate the pain of patients suffering from vertebral pain.

RESULTSAll the patients had suffered from vertebral pain before treatment. VAS was 9 points in 4 cases, 8 points in 6 cases, 7 points in 1 case, 3 points in 6 cases, and 2 points in 3 cases. Among them, 7 patients complicated with neurological symptoms, 11 with aggravating night pain, 10 with fever. WBC and Neutrophil count (NEU) of 5 cases were increased and other 15 cases were normal;CRP of 19 cases were increased and 1 case was normal;ESR of all 20 cases were increased. X rays showed the intervertebral space narrowing in all 20 cases, 13 cases complicated with destruction of vertebral body; CT showed the lesions of vertebral body in the 20 cases and complicated with destruction, sclerosis of sclerotin; MRI showed that the lesions of the vertebral body in the T1 image had uneven medium low signal, in the T2 image of the 16 cases had uneven high signal and 2 cases had uniform and high signal, 2 cases had main high signal compliated with mixed signal. Thirteen patients underwent surgical treatment and 7 patients received conservative treatment, and the patients left hospital while VAS had significantly improved after treatment.

CONCLUSIONSPyogenic spondylitis is easy to be misdiagnosed or missed in clinic. It can be combined with the clinical manifestations, laboratory examination and imaging characteristics in order to make a definite diagnosis for purulent spondylitis in early.

Objective:To investigate the influence of rosuvastatin combined ticagrelor on therapeutic effect,cardiac func-tion and inflammatory factor levels in patients with acute myocardial infarction(AMI).Methods:A total of 288 AMI pa-tients admitted in our hospital were randomly and equally divided into rosuvastatin group(received rosuvastatin therapy)and combined treatment group(received rosuvastatin combined with ticagrelor).After two-month treatment,therapeu-ticeffect,cardiac function,inflammatory index levels and incidence rate of adverse reaction were compared between two groups.Results:Total effective rate of combined treatment group was significantly higher than that of rosuvastatin group(96.53%v.s 86.81%,P=0.003).Compared with before treatment,there were significant improvements in cardiac function and inflammatory factor levels in two groups after treatment,P=0.001 al.l Compared with rosuvastatin group af-ter treatment,there were significant reductions in left ventricular end-diastolic diameter(LVEDd)[(44.86±5.25)mmv.s(30.94±4.02)mm],left ventricular end-systolic diameter(LVESd)[(28.83±3.02)mmv.s(21.27±2.86)mm],interventricular septal thickness(IVST)[(9.82±1.51)mmv.s(9.09±1.46)mm],levels of interleukin-6(IL-6)[(4.62±0.68)pg/mlv.s(3.85±0.59)pg/ml]and tumor necrosis factor-α(TNF-α)[(1.25±0.22)pg/mlv.s(0.93±0.18)pg/ml],and significant rise in 6 min walking distance(6MWD)[(398.63±50.58)m v.s(436.35±52.03)m]and LVEF[(54.11±6.32)%v.s(60.24±7.88)%]in combined treatment group,P=0.001 al.l There was no signif-icant difference in total incidence rate of adverse reactions during treatment between two groups,P=0.444.Conclusion:The combination of rosuvastatin and ticagrelor possesses significant therapeutic effect in the treatment of AMI patient.s It can improve cardiac function and reduce levels of inflammatory factors with good safety.