Adult ; Antibodies, Monoclonal ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Follow-Up Studies ; Humans ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; Male ; Mucin-1 ; metabolism ; Prednisone ; therapeutic use ; Receptor Protein-Tyrosine Kinases ; metabolism ; Vincristine ; therapeutic use

To investigate the potential ability of the nitrite to induce neuronal differentiation of PC12 cells, cultured PC12 cells planted on matrigel in the presence or absence of sodium nitrite were employed as model, nerve growth factor (NGF) served as a positive control. After 48 h, sodium nitrite enhanced cell viability and vascular endothelial growth factor (VEGF) secretion. Same as the effect of NGF, sodium nitrite (1.4 mmol x L(-1)) treated cultures contained a greater proportion of cells bearing neurites and neurites were much longer than those found in negative control cultures (P < 0.05). Compared with the negative control, sodium nitrite (1.4 mmol x L(-1)) also upregulated the expression of VEGF mRNA (P < 0.05) and hypoxia inducible factor 1 alpha (HIF-1 alpha) or VEGF protein expression (P < 0.05) in cultures of PC12 cells. On the other hand, these effects of the sodium nitrite were likely mediated by HIF-1alpha, since their effects were antagonized by addition of HIF-1alpha inhibitor YC-1. Taken together, these results suggest that low doses of sodium nitrite could induce neurite outgrowth in PC12 cells by activating the HIF-1alpha-VEGF pathway.
Animals ; Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Food Preservatives ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Neurites ; drug effects ; PC12 Cells ; RNA, Messenger ; metabolism ; Rats ; Sodium Nitrite ; pharmacology ; Up-Regulation ; Vascular Endothelial Growth Factor A ; genetics ; secretion

This study is to find out the induction by sodium nitrite of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma cells, SMMC-7721. After treatment of SMMC-7721 with 0.25 - 25 mmol.L-1 sodium nitrite for 48 h, the assays used include enzyme-linked immunosorbent assay (ELISA) for evaluation of TGF-beta1, IL-6 and IL-8 level in the conditioned medium, phase-contrast microscopy for morphology observation, and scratch wound healing as well as transwell migration assays for measurement of migration and metastatic potential. Additionally, the hallmarks of EMT, p-AKT and its downstream signaling molecules were examined by Western blotting. The results showed that TGF-beta1 secreted by SMMC-7721 elevated significantly in a dose-dependent fashion, whereas the increased IL-8 and IL-6 did not show dose-dependent response. The EMT was induced by exposure of SMMC-7721 with 0.25 mmol.L-1 of sodium nitrite, which was characterized by increased level of Vimentin, decreased E-cadherin and elevated activity of migration and metastatic potential. The results suggest that sodium nitrite could induce SMMC-7721 EMT by increased secretion of TGF-beta1 and IL-8.
Cadherins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Dose-Response Relationship, Drug ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Liver Neoplasms ; metabolism ; pathology ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-akt ; metabolism ; Sodium Nitrite ; administration & dosage ; pharmacology ; Transforming Growth Factor beta1 ; secretion ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism

This study is to observe the effect of N-(3-phenylallylidene)-6-fluoro-1, 8-(2, 1-propoxy)-7-(4-methylpiperazin-1-yl)-quinolin-4(1H)-one-3-carbonyl hyarazine (FQ16) on apoptosis of hepatocarcinoma SMMC-7721 cells in vitro. With different concentrations of FQ16 at different times used to treat SMMC-7721 cells in vitro, the proliferation of the cells and the inhibition effect of FQ16 on the cell proliferation were examined by MTT assay. Cell apoptosis was determined by Hoechst 33258/PI fluorescence staining, TUNEL and agarose gel electrophoresis method. The effect of FQ16 on topoisomerase II activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Mitochondrial membrane potential (MMP, delta psi m) was measured by high content screening image system. The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression changes of Bcl-2 mRNA and Bax mRNA. The caspase-9, caspase-8, caspase-3, p53, Bcl-2 and Bax protein expressions were detected by Western blotting analysis. The results showed that the cell proliferation was inhibited by FQ16 at 0.625 - 10 micromol L(-1) in a time-dose dependent manner. Treatment of SMMC-7721 cells with different concentrations of FQ16 for 24 h increased the percentage of the apoptosis cells obviously (P<0.05), the typical ladder DNA in apoptotic cells and a concomitant dissipation of the mitochondrial membrane potential. Compared with control group, FQ16 influenced obviously DNA topoisomerase II activity, stimulated DNA cleavage and inhibited DNA reunion mediated by topoisomerase II. In addition, FQ16 (3 - 7.39 micromol L(-1)) increased mRNA expression of Bax and protein expression of p53, Bax, caspase-9, caspase-3, separately, and induced cytosolic accumulation of activities caspase-9 and caspase-3, whereas the mRNA and protein expression of Bcl-2 decreased with no change of caspase-8. Therefore it can be concluded that the effects of inhibited topoisomerase II and mitochondrial-dependent pathways were involved in FQ16 induction of apoptosis of SMMC-7721 cells.
Antineoplastic Agents ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type II ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Molecular Structure ; Piperazines ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism

This study is to investigate the cytoprotective role of NaNO2 preconditioning against ethanol induced damage in human hepatoma SMMC-7721 cells. The cells were preconditioned with NaNO2 (0.25 mmol x L(-1)) for 24 hours or 4 weeks, and then exposed to ethanol (200 mmol x L(-1)) for additional 12 h and untreated cells served as control. Both temporal and chronic NaNO2 preconditioning could prevent ethanol elicited cytotoxicity as evidenced by thiazolyl blue (MTT). NaNO2 preconditioning also could inhibit ethanol-induced apoptosis, which was confirmed by FITC-Annexin V/PI flow cytometer and Hoechst 33258 and PI staining. Further, simultaneous NaNO2 preconditioning treatment along with ethanol showed protection against ethanol mediated cellular damage as indicated by significantly decreased levels of malondialdehyde (MDA) and elevated activities of superoxide dismutase (SOD) and catalase (CAT). Western blotting analysis revealed that in ethanol treated cells preconditioned with NaNO2, the HIF-1alpha and Bcl-2 increased obviously, while the expression of pro-apoptotic proteins, including Bax, Caspase-9, Caspase-3 decreased. The results showed that low doses of NaNO2 preconditioning resistant to ethanol-induced human hepatoma SMMC-7721 cells apoptosis, which mechanism may be related to increased expression of HIF-1alpha in the cells.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Catalase ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ethanol ; toxicity ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Malondialdehyde ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sodium Nitrite ; administration & dosage ; pharmacology ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; metabolism

The aim of this study was to determine the correlation of ultrasonographic estimates of testicular volume with true testicular volume and to compare the accuracy and precision of the three most commonly utilized formulas. A total of 15 patients underwent high-resolution ultrasonography (US) analysis for testicular volume before orchiectomy. Testicular volume was calculated using three common formulas: (1) length (L) x width (W) x height (H) x 0.52; (2) the empirical formula of Lambert: L x W x H x 0.71; and (3) L x W2 x 0.52. The actual volume of each removed testis was estimated directly by a water displacement method. Thus, four volume measurements were obtained for each of the 30 testes. The obtained data were analyzed by paired t-test and linear regression analysis. All three US formula measurements significantly underestimated the true testicular volume. The largest mean biases were observed with US formula 1, which underestimated the true volume by 3.3 mL (31%). US formula 2 had a smaller mean difference from the true volume, with an underestimation of only 0.6 mL (6%). Regression analysis showed that formulas 1 and 2 had better R2 values than formula 3. However, all three US formulas displayed a strong linear relationship with the true volume (R2= 0.872-0.977; P < 0.001). Among the commonly used US formulas, the empirical formula of Lambert (L x W x H x 0.71) provided better accuracy than the other two formulas evaluated, and better precision than formula 3. Therefore, the formula of Lambert is the optimal choice in clinical practice.
Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Organ Size ; Reproducibility of Results ; Testis ; anatomy & histology ; diagnostic imaging ; Ultrasonography ; methods

OBJECTIVETo sought to engineer and characterize a biodegradable nanoparticles (NPs) containing rapamycin which use poly (lactic-co-glycolic) acid (PLGA) as the carrier matrix and to assess its in vivo release characteristics by local drug delivery system intravascularly.

METHODSRapamycin-loaded PLGA NPs were prepared by an emulsification/solvent evaporation technique, and NPs size distribution was assessed by submicro laser defractometer. The particle morphology was observed by scanning electron microscopy. In vitro release from the NPs was performed in TE buffer at 37 degrees C under rotation utilizing double-chamber diffusion cells on a shake stander. In vivo NPs intravascular local delivery were performed by DISPATCH catheter in New Zealand rabbit abdominal aorta and Chinese experimental mini-pigs coronary artery models.

RESULTSBiodegradable rapamycin loaded PLGA NPs were constructed successfully by emulsification solvent-evaporation technique. The diameter of rapamycin-PLGA NPs was around 246.8 nm with very narrow size distribution, and rapamycin-NPs showed good spherical shape with smooth uniform surface. Rapamycin loaded in NPs were around was 19.42%. Encapsulation efficiency of drug was over 77.53%. The in vitro release of rapamycin from NPs showed that 75% of the drug was sustained released over 2 weeks and controlled release in a linear pattern. After a single 10 minutes infusion of rapamycin-PLGA NPs suspension (5 mg/ml) under 20.27 kPa through DISPATCH catherter in vivo, the mean rapamycin levels at 7 day and 14 day were (2.438 +/- 0.439) and (0.529 +/- 0.144) microg/mg of the dry-weight of the artery segments (2 cm) which local delivery were administrated.

CONCLUSIONSPLGA NPs controlled drug delivery system for intraarterial local anti-proliferative drug delivery can potentially improve local drug concentration and prolong drug residence time in animal model in vivo. It should be appropriate for further study of its therapy efficiency in human.

Animals ; Aorta, Abdominal ; drug effects ; Coronary Vessels ; drug effects ; Drug Carriers ; chemistry ; Drug Delivery Systems ; methods ; Infusions, Intra-Arterial ; Lactic Acid ; chemistry ; Nanoparticles ; chemistry ; Particle Size ; Polyglycolic Acid ; chemistry ; Rabbits ; Sirolimus ; administration & dosage ; pharmacokinetics ; Swine ; Swine, Miniature

BACKGROUNDAtherosclerosis is a chronic inflammatory disease. Accumulated evidences suggest a deep involvement of oxidative damage in the development of atherosclerosis, but little is discussed over the relationship between plasma glutathione redox status as the most important intrinsic antioxidant defensive mechanism and the atherosclerosis.

METHODSA total of 132 patients suspected with atherosclerosis were assigned to three groups by high frequency ultrasonic examination of the carotid artery. With the thickness of intima of the carotid artery as an index of degree of atherosclerosis progression, 56 were included in plaque-forming group (A), 42 in carotid artery intima-thickening group (B), and 34 in normal carotid artery intima-thickness group (C). All patients were subjected to the measurement of plasma glutathione (GSH) (reduced form GSH and oxidized form GSSG), nicotinamide adenine dinucleotide phosphate (NADP) (reduced form NADPH and oxidized form NADP(+)), oxidized low density lipoprotein (oxLDL), and malondialdehyde (MDA). The GSH/GSSG and NADPH/NADP(+) redox potentials were calculated according to the Nernst equation, and their correlation with intima thickness and oxLDL was analyzed.

RESULTSWith the thickening of artery intima (from group C to A), GSH concentration and the ratio of GSH/GSSG gradually reduced, and GSSG and GSH/GSSG redox potential gradually increased (more positive) (P < 0.05). The NADPH and NADPH/NADP(+) redox status also showed similar but milder changes. The products of oxidative stress oxLDL and MDA increased significantly along with the thickening of artery intima (P < 0.05). The analysis of the relationship between GSH/GSSG redox potential, intima thickness, and oxLDL showed positive correlations (P < 0.05). The plasma GSH/GSSG redox status was positively correlated with the intima thickness of the carotid artery and the oxidized injury of LDL. The redox status shifted to oxidizing direction along with the intima thickening and plaque-forming.

CONCLUSIONElevated peroxidative glutathione redox status was deeply implicated in atherosclerosis progressing, and it may be a sensitive and reliable index for monitoring oxidative status in atherosclerosis.

Adult ; Aged ; Atherosclerosis ; metabolism ; pathology ; Carotid Arteries ; pathology ; Female ; Glutathione ; blood ; Glutathione Disulfide ; blood ; Humans ; Lipid Peroxidation ; Male ; Middle Aged ; NADP ; blood ; Oxidative Stress ; Tunica Intima ; pathology ; Tunica Media ; pathology

OBJECTIVETo investigate the distribution of the Beijing genotypes of Mycobacterium tuberculosis (M. tuberculosis) and the relationships between Beijing genotype strains and drug-resistant phenotypes in China.

METHODSClinical isolates were collected during a 9-month research period from April to December in 2008 in six geographic regions of China. One isolate that had been biochemically confirmed to be a member of the M. tuberculosis complex was collected from each patient. The demographic data of the patients (eg. sex, age, and history of tuberculosis) as well as the drug resistance patterns and sources of the clinical isolates were collected. Drug susceptibility testing was performed using proportion method. Beijing genotypes of M. tuberculosis were identified by spacer oligonucleotide typing or insertion of IS6110 in the genomic dnaA-dnaN locus.

RESULTSAmong the 410 M. tuberculosis clinical isolates, 67.1% (275/410) isolates were Beijing genotypes of M. tuberculosis. Significantly larger proportions of tuberculosis patients were infected with Beijing genotypes in the northeastern regions of China than that of in the central-western regions (chi2 = 20.50, P = 0.000). No significant associations were found either between Beijing genotype strains and patients' age, sex, or treatment history. Multidrug-resistant isolates and rifampin-resistant isolates were more common among Beijing genotype strains than among non-Beijing strains (P = 0.002, P = 0.005).

CONCLUSIONSAbout two third of the clinical isolates of M. tuberculosis in China are Beijing genotypes. Beijing genotype strains are not correlated with patients' age, sex, treatment history. People living in the northeastern regions of China are more susceptible to Beijing genotypes than those living in the central-western of China. Beijing genotype strains tend to be rifampin-resistant or multidrug-resistant.

Antitubercular Agents ; pharmacology ; China ; Genotype ; Humans ; Mycobacterium tuberculosis ; classification ; genetics ; Phenotype ; Rifampin ; pharmacology ; Tuberculosis ; drug therapy ; Tuberculosis, Multidrug-Resistant ; genetics

Objective:To investigate the clinical efficacy of membrane induction technique in the treatment of postoperative infection of tibial plateau fractures in adults.Methods:A retrospective case series study was conducted to analyze the clinical data of 21 adult patients with postoperative infection of tibial plateau fractures treated with membrane induction technique from April 2013 to May 2017 in Southwest Hospital of Army Medical University. There were 19 males and two females, aged 19-60 years [(44.1±5.8)years]. There was one patient with type IV fractures, 14 with type V, and 6 with type VI according to the initial fracture typing by Schatzker's classification. There were three patients with infection period of within 3 weeks, 12 of 3-10 weeks, and 6 of over 10 weeks. All patients underwent two-stage operation using membrane induction technique to place cement in the bone defect area. After removal of internal fixation and thorough debridement, antibiotic cement and internal fixation plate were placed at stage I. Bone graft and reconstruction was performed at stage II. The infection indicators were recorded. Infection indices were monitored, including white blood cell count (WBC), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). Clearance of infection, bony union and complications were evaluated. Range of motion (ROM) and scoring of Hospital for Special Surgery (HSS) were used to evaluate the function of knee joint.Results:All patients were followed up for 12-62 months with an average of 23.5 months. Compared with 3 months after stage II, the indicators of infection at stage I showed that WBC was decreased from (10.6±2.3)×10 9/L to (6.7±3.5)×10 9/L, ESR decreased from (26.0±5.3)mm/h to (12.1±4.3)mm/h, and CRP decreased from (10.0±1.5)mg/L to (5.8±1.0)mg/L ( P<0.05). Infection was cleared in 17 patients after stage I operation, and the other 4 patients had infection recurrence, which were given stage I debridement again to control the infection. Two patients were treated with local flap transfer to cover the wound because of skin soft tissue defect after debridement. Another two patients underwent knee arthrodesis, and none was amputated. X-ray film indicated bony union in 21 patients at 46 months (mean, 4.5 months) after operation, and clinical bone healing was acquired in all 21 patients. One patient showed donor site infection. No nonunion, recurrence of infection after stage II, deep vein thrombosis or pulmonary embolism occured after the second stage. At the latest follow-up, ROM in patients with infection periods within 3 weeks and 3-10 weeks was singnificantly improved from [(95.2±10.4)° and (85.7±11.5)°] to [(120.2±10.5)° and (98.6±12.2)°] ( P<0.01), but not in patients with infection periods of over 10 weeks ( P>0.05). The HSS score in all patients was significantly improved after operation [(65.6±8.2)points vs. (82.0±6.6)points]( P<0.01). Conclusion:For adult patients with tibial plateau fracture, membrane induction technique can effectively control the postoperative infection, achieve clinical bone healing and improve the knee function.