OBJECTIVETo compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment.

METHODSTwenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen type III, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture.

RESULTSHE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group,but there was severely degenerated of 21 days segment group. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group (P > 0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days (P < 0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups (P > 0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P > 0.05) however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased,and there was a significant difference compared with before each time points and organ group at 21 days (P < 0.05).

CONCLUSIONIt is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebral disc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days.

Animals ; Collagen Type II ; analysis ; Female ; Immunohistochemistry ; Intervertebral Disc ; chemistry ; pathology ; Male ; Organ Culture Techniques ; Rabbits

Objective To evaluate the clinical outcome and relative factors of intervention treatment for atherosclerotic renal artery stenosis in elderly patients.Methods The clinical data of 79 patients diagnosed as atherosclerotic renal artery stenosis by angiography and treated by revascularization were analyzed.Results There were 55(69.6%)successes and 24(30.4%)failures in decreasing blood pressure and 28(35.4%)successes and 51(64.6%)failures in improving renal function after intervention treatment.Predictors of favorable outcome of intervention treatment in decreasing blood pressure were related to lower urine protein,higher glomerular filtration rate,higher systolic and diastolic blood pressure before treatment,lower resistance index(RI)of renal artery,and no complication of cerebral vascular diseases.Predictors of favorable outcome of intervention treatment in improving renal function were related with percentage of angiographic stenosis,category of antihypertension and lower urine protein.The logistic regression analysis showed that the percentage of angiographic stenosis was the most important predictor of intervention treatment for blood pressure control,age and level of serum creatinine before intervention treatment were the most important predictors of intervention treatment for improving renal faction.Conclusion Percentage of stenosis(≥85%),age(133 ?mol/L)can be used as the predictors of therapeutic success for renovascular stenosis in older patients.

Objective To investigate the clinical value of serum metabonomic profile of pancreatic cancer using nuclear magnetic resonance (NMR)-based metabonomics.Methods The retrospective case-control study was adopted.The clinical data of 23 patients with pancreatic cancer (PC group) and 16 healthy volunteers (control group) who were admitted to the Fujian Medical University Union Hospital between December 2013 and December 2014 were collected.The serum of the 2 groups was measured by 1H NMR spectroscopy.Multivariate statistical analyses were performed to identify the characteristic metabolites in the 2 groups,including principal component analysis (PCA),partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA).Observation indicators included:(1) multivariate statistical analysis of serum metabonomic profile,results of PCA,PLS-DA and OPLS-DA,(2) screening of metabolites.Measurement data with normal distribution were presented as x ± s.The comparison between groups was evaluated with the t test.The count data were analyzed using the chi-square test.Results (1) The multivariate statistical analysis of serum metabonomic profile:results of PCA showed that expression rates of principal component 1 (PC1) and principal component 2 (PC2) to original data were 54.9％ and 23.5％,with both cumulative contribution rate of 78.4％.Results of PLS-DA showed that the separative trend between PC group and control group was appeared,and variance of X and Y matrixes and predictive value were 0.254,0.816 and 0.385.Results of OPLS-DA showed that the differences of samples between the 2 groups were further increased,and differential metabolites were screened according to the distinction of scores between the 2 groups,value of R2X,R2Y and Q2 was 0.254,0.816 and 0.433.(2) Screening of metabolites:35 serum metabolites were detected in the 2 groups.Compared with the control group,levels of 3-hydroxybuyarate,citrate,formate,glutamate,isoleucine,methionine and phenylalanine in the PC group were elevated (r =0.524,0.511,0.656,0.566,0.503,0.498,0.648,P ＜0.05),and levels of 3-methylhistidine,alanine,glutamine,LDL and VLDL in the PC group were decreased (r =-0.607,-0.508,-0.560,-0.568,-0.559,P ＜ 0.05).Conclusions Compared with healthy controls,several amino acids,citrate and lipoproteins demonstrate the metabolic differences in the serum of patients with pancreatic cancer.NMR based metabonomic profile technology can distinguish the difference of serum metabolites between patients with pancreatic cancer and healthy controls.NMR based metabonomic technology may be a promising method for the diagnosis of pancreatic cancer.

Fourteen compounds were isolated from Dalbergia odoriferae and purified by repeated column chromatography on silica and sephadex LH-20 gel and structurally identified by spectral analysis. These compounds were identified as 4, 9-dimethoxy-3-hydroxypterocarpan (1), medicarpin (2), 2', 4', 5-trihydroxy-7-methoxyisoflavone (3), 2', 3', 7-trihydroxy-4'-methoxyisoflavan (4), formononetin (5), 3, 8-dihydroxy-9-methoxypterocarpan (6), koparin (7), 3-hydroxy-9-methoxypterocarp-6a-ene (8), 2'-hydroxyformononetin (9), stevenin (10), 2', 7-dihydroxy-4', 5'-dimethoxyisoflavone (11), lyoniresinol (12), 2, 4-dihydroxy-5-methoxy-benzophenone (13) and neokhriol A (14). Compounds 1, 3, 4, 6, 8, 12 and 14 were isolated from this plant for the first time. Antibacterial activity assay showed that compound 4 had inhibitory effect on Ralstonia solanacearum.
Anisoles ; chemistry ; isolation & purification ; pharmacology ; Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Benzophenones ; chemistry ; isolation & purification ; pharmacology ; Chromatography ; methods ; Dalbergia ; chemistry ; Dextrans ; Gels ; Isoflavones ; chemistry ; isolation & purification ; pharmacology ; Microbial Sensitivity Tests ; Naphthalenes ; chemistry ; isolation & purification ; pharmacology ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Pterocarpans ; chemistry ; isolation & purification ; pharmacology ; Ralstonia solanacearum ; drug effects ; growth & development ; Silica Gel

Objective To evaluate the stability and biodistribution of a novel SPECT-MRI bi-functional agem 99Tcm-Gd-DTPA-DG in tumor-bearing nude mice. Methods DTPA-DG was synthesized and then conjugated with Gd2O3 to generate Gd-DTPA-DG. The tumor-bearing nude mice were scanned by MRI to evaluate the tumor targeting ability of Gd-DTPA-DG. The orthogonal experiment was applied to optimize pH value of reaction medium and reaction temperature. The radiolabeling efficiency was measured by thin layer chromatography. The distribution of 99Tcm-Gd-DTPA-DG in nude mice was evaluated by scintigrapy in vivo. The % ID/g was measured at different time after intravenous injection of 99Tcm-Gd-DTPA-DG. Results The tumor was significantly enhanced by Gd-DTPA-DG with MRI. The radiochemical purity of 99Tcm-Gd-DTPADG was about 98.5% and remained 96.2% at room temperature for 6 h. The tumor was well visualized by 99TcmGd-DTPA-DG SPECT at 2 h after injection. The tumor uptake was (1.48 ±0.12) %ID/g, and the rumor to muscle radioactivity ratio was 2.91. Conclusions MRI contrast of Gd-DTPA-DG may enhance tumor detection. 99Tcm-labeled Gd-DTPA-DG may be useful for tumor imaging and might have a potential role as a SPECT-MRI bi-functional agent.

OBJECTIVETo investigate the role of vancomycin-loaded calcium sulphate in the treatment of osteomyelitis.

METHODSWe implanted vancomycin-loaded calcium sulphate into 24 patients with traumatic osteomyelitis who were treated in our hospital from February 2008 to February 2010,and then the antibiotic concentrations in the lesions were measured.Bacterial culture results,inflammatory markers,as well as wound healing were observed.X-ray was performed in the location where the vancomycin-loaded calcium sulphate was implanted.The blood vancomycin level as well as liver and kidney functions were determined.

RESULTSThe vancomycin concentration in the lesion exceeded the effective therapeutic concentrations and the minimum inhibitory concentration,while the blood concentration was low.The liver and renal function remained normal.The safety profile was good,and the cure rate of osteomyelitis reached 100%.

CONCLUSIONSThe vancomycin-loaded calcium sulphate can release high-concentration vancomycin in the diseased sites without causing high blood concentration.Also,it can guide the regeneration of bones.Therefore,it is effective and safe in treating osteomyelitis.

Adolescent ; Adult ; Anti-Bacterial Agents ; therapeutic use ; Biocompatible Materials ; Calcium Sulfate ; therapeutic use ; Child ; Drug Carriers ; Female ; Humans ; Male ; Middle Aged ; Osteomyelitis ; therapy ; Vancomycin ; pharmacokinetics ; therapeutic use ; Young Adult

AIMTo study the distribution and efficiency of pEGFP-N1 plasmid after its transfection in cardiomyocytes.

METHODSThe neonatal rat cardiomyocytes were cultured. According to the different grow period of neonatal rat cardiomyocytes, the distribution and efficiency of pEGFP-N1 plasmid after its transfection in cardiomyocytes were studied.

RESULTSThe efficiency was significantly increased after pEGFP-N1 plasmid transfection in one-day cardiomyocytes. The distribution of EGFP was in the cytoplasm and nucleus.

CONCLUSIONThe efficiency of pEGFP-N1 plasmid after its transfection in cardiomyocytes was related to the grow period of neonatal rat cardiomyocytes. The distribution of EGFP was in the cytoplasm and nucleus.

Animals ; Animals, Newborn ; Cells, Cultured ; Green Fluorescent Proteins ; genetics ; metabolism ; Myocytes, Cardiac ; metabolism ; Plasmids ; Rats ; Rats, Wistar ; Transfection

Adult ; Asian Continental Ancestry Group ; DNA, Viral ; blood ; Female ; Genes, Viral ; genetics ; Hepatitis B virus ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Protein Isoforms ; genetics ; Viral Core Proteins ; genetics

The plaque-forming characteristics of Newcastle disease viruses of chickens and geese source were compared on various cells. The result showed that there were obvious differences of plaque formation between F48E9 and NA-1 on Vero cells, chicken embryo fibroblast cells (CEF) and goose embryo fibroblast cells (GEF). The plaque-forming ability of NA-1 was higher than F48E9 on GEF, but lower than F48E9 on CEF. On Vero cells, the plaque-forming ability of NA-1 was slightly stronger than F48E9. It demonstrated that the plaque-forming characteristics were consistent with host tropism of virus. The morphogenesis of F48E9 and NA-1 on Vero cells was observed with transmission electron microscope. There were different replication processes between F48E9 and NA-1 on cells at different stages. NA-1 had stronger adaptability to host than F48E9 according to budding processes and envelope integrity.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Geese ; Host-Pathogen Interactions ; Newcastle Disease ; virology ; Newcastle disease virus ; growth & development ; isolation & purification ; physiology ; ultrastructure ; Poultry Diseases ; virology ; Vero Cells ; Viral Plaque Assay

OBJECTIVETo investigate the effect of the selective PI3K inhibitor and MEK inhibitor on KRAS and PTEN co-mutated non-small cell lung cancer cell line NCI-H157 and the relevant mechanisms.

METHODSNCI-H157 was cultured routinely and treated with different concentrations of the two inhibitors. Cell proliferation was detected by MTT cell cycle assay. Based on the MTT results the cells were divided into four groups: the control group, PI3K inhibitor group (GDC-0941, 0.5 and 5.0 µmol/L), combination group I (0.5 µmol/L AZD6244 + 0.5 µmol/L GDC-0941) and combination group II (5.0 µmol/L AZD6244 + 5.0 µmol/L GDC-0941). Colony formation assay was performed to detect colony formation efficiency. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of protein related to apoptosis was tested with Western blot.

RESULTSCell growth was inhibited by the two inhibitors. Combination groups led to stronger cell proliferation inhibition: combination group Ishowed synergistic effect of their actions and combination group II showed an additive effect; in both groups, there were decreased colony number [(77.2 ± 1.54)/well vs (61.50 ± 2.12)/well, P < 0.01] and [(51.00 ± 4.00)/ well vs (22.50 ± 3.53)/well, P < 0.01]; and enhanced apoptotic ratios [(18.30 ± 0.82)% vs (21.32 ± 0.56)%, P < 0.01] and [(27.14 ± 1.58)% vs (42.45 ± 4.42)%, P < 0.01]. In addition, compared to the PI3K inhibitor alone group, the NCI-H157 cells in the combination groups showed increased G0/G1 phase and decreased S phase (P < 0.01). Western blotting showed that the combination groups demonstrated significantly decreased expression of cyclin D1 and cyclin B1, increased p21 and cleaved PARP and decreased bcl-2/bax ratio, compared to the PI3K inhibitor only group.

CONCLUSIONThe combined inhibition of PI3K (AZD6244) and MEK (GDC-0941) has synergistic effects on the proliferation of NCI-H157 cells, but such effects appear to be in a dose-dependent manner.

Apoptosis ; drug effects ; Benzimidazoles ; administration & dosage ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin B1 ; metabolism ; Cyclin D1 ; metabolism ; Dose-Response Relationship, Drug ; Drug Synergism ; Humans ; Indazoles ; administration & dosage ; pharmacology ; Lung Neoplasms ; genetics ; pathology ; Mitogen-Activated Protein Kinase Kinases ; antagonists & inhibitors ; metabolism ; Mutation ; PTEN Phosphohydrolase ; genetics ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; metabolism ; Poly(ADP-ribose) Polymerases ; metabolism ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins p21(ras) ; metabolism ; Signal Transduction ; Sulfonamides ; administration & dosage ; pharmacology ; bcl-2-Associated X Protein ; metabolism ; ras Proteins ; genetics