OBJECTIVETo investigate whether RhoA/Rho-kinase contributes to the occurrence of chronic post-thoracotomy pain (CPSP) by up regulation of glutaminase 1 (GLS1) expression in the spinal dorsal cord.

METHODSTwenty five male Sprague Dawley (SD) rats were divided into control group (n=5) and model group (n=20). The rats in the model group were randomized into two sub groups (n=10) for observation on day 10 and day 21 after thoracotomy, and each group was further divided into CPSP and non CPSP groups according to the behavioral test results. All the rats were sacrificed after behavioral test for examination of GLS1 and RhoA expressions in the spinal cord using Western blotting and RT PCR. We also compared the effect of the Rho kinase inhibitor fasudil and saline, both injected intraperitoneally daily at 10 mg/kg for 7 consecutive days following thoracotomy, on CPSP and GLS1 expression in 30 male SD rats on day 21 after thoracotomy.

RESULTSCompared with the control group, the rats with CPSP showed significantly increased expressions of GLS1 and RhoA mRNA in the spinal cord on both day 10 and day 21 following thoracotomy (P<0.01), but the rats without CPSP did not show obvious changes in GLS1 and RhoA expressions. In fasudil treated rats, the mechanical pain threshold was obviously increased and the expressions of GLS1 and RhoA were significantly reduced as compared with those in saline treated rats (P<0.01).

CONCLUSIONRhoA plays an important role in the occurence of CPSP by up-regulating the expression of GLS1 in the spinal dorsal cord of rats.

OBJECTIVETo investigate the expressions of vascular endothelial growth factors (VEGF)-A, -C and -D and their prognostic significance and relation to angio- and lymphangiogenesis in gastric cancer.

METHODSThe expression of VEGF-A, -C and -D in 123 primary gastric cancers was detected by immunohistochemical staining. The lymphatic vessel density (LVD) and microvessel density (MVD) were assessed after immunohistochemical double-staining with D2-40 and CD34, respectively. The correlation between the expression of those VEGF factors and clinicopathological parameters were analyzed by univariate method. The overall survival was evaluated by Kaplan-Meier method and log-rank test. Multivariate analysis was carried out using Cox proportion hazard model.

RESULTSThe positive expression rate of VEGF-A, -C and -D in primary gastric cancer samples were 64.2%, 65.9% and 41.5%, respectively. High expression of VEGF-A, or -C or -D, or any two of them was correlated with high LVD (P < 0.05). High expression of both VEGF-A and -C was associated with high MVD, lymph node metastasis, LVI and MVI (P < 0.05). Both VEGF-C and -D high expression was correlated with LVI and lymph node metastasis (P < 0.05). The patients with high expression of these factors had a statistically shorter overall survival (P < 0.05). The patients with both VEGF-A and -C expression had the shortest survival (56 months). Multivariate analysis showed that VEGF-A high expression, MVD, lymph node metastasis and depth of tumor invasion were independent survival predictors (P = 0.033, 0.002, 0.019 and P < 0.001, respectively).

CONCLUSIONHigh expression of both VEGF-A and -C imply high potential of lymphangiogenesis, metastasis and poorer survival in gastric cancer patients. High expression of VEGF-C and -D may induce lymphangiogenesis and promote lymph node metastasis, but only VEGF-A is an independent predictor of survival.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Microvessels ; pathology ; Middle Aged ; Neovascularization, Pathologic ; Proportional Hazards Models ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVETo investigate the susceptibility of Streptococcus mutans (S. mutans) biofilms to antimicrobial agent by confocal laser scanning microscopy (CLSM).

METHODSS. mutans biofilms formed in vitro on glass slice were acted on with penicillin of different concentrations for 3 h. Then these biofilms were stained by fluorescence and were observed by CLSM. The bacterial density and viability of biofilms were recorded.

RESULTSWhen S. mutans biofilms were exposed to penicillin of 2 500 mg/L for 3 h, it was not completely killed. The higher the concentration of penicillin was, the weaker the biofilms against penicillin.

CONCLUSIONSCompared with planktonic S. mutans, S. mutans biofilms produced stronger resistance to penicillin. It suggests that we should find new strategies to control the infection caused by biofilm in clinic.

Anti-Infective Agents ; administration & dosage ; pharmacology ; Biofilms ; drug effects ; Dose-Response Relationship, Drug ; Microbial Sensitivity Tests ; Microscopy, Confocal ; Penicillins ; administration & dosage ; pharmacology ; Streptococcus mutans ; drug effects

BACKGROUNDTo investigate the frequency of circulating HBV specific T helper cell and evaluate its association with serum levels of HBV DNA before and during lamivudine treatment in patients with chronic hepatitis B.

METHODSThe frequency of circulating HBV specific T helper cells in response to HBcAg in 25 chronic HBV-infected patients was determined by Elispot assay; serum HBV DNA was quantitated by real-time PCR.

RESULTSThe frequency of HBV specific T helper cell before antiviral treatment (47.30 +/- 25.50 SFCs /1 x 10(6) PBMC) was significantly higher than that at the third month of therapy (23.10 +/- 18.45 SFCs /1 x 10(6) PBMC, P < 0.05). All 8 patients observed dynamically had decreased frequency of HBV specific T helper cell at the third month of therapy; six patients with serum HBV DNA level reduced had higher frequency of HBV specific T helper cell before treatment than 2 patients without serum HBV DNA level decrease.

CONCLUSIONHBV specific T helper cell response at the time of hepatitis flare in chronic hepatitis B patients was significantly augmented compared to that at the time of catabasis.

Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Hepatitis B Core Antigens ; immunology ; Hepatitis B virus ; drug effects ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Male ; T-Lymphocytes, Helper-Inducer ; cytology ; drug effects ; immunology

OBJECTIVETo compare the inhibitory effects of cytokine-induced killer (CIK) cells alone, chemotherapeutic drug alone, and CIK cells combined with chemotherapeutic drug on the growth of hepatocellular carcinoma (HCC) cells transplanted in nude mice.

METHODSPeripheral blood mononuclear cells (PBMC) collected from five healthy donors by blood cell separator were incubated in vitro to induce CIK cells in the presence of interferon-gamma (IFN-gamma), IL-2 and anti-CD3 monoclonal antibody (mAb). The phenotype of CIK cells was characterized by flow cytometric analysis. BEL-7402 HCC cells were inoculated subcutaneously to nude mice. On day 5, at the inoculation site were injected normal saline (group 1), CIK cells (3 x 10(7) and 6 x 10(7), group 2 and 3), mitomycin-C (MMC 80 microg in 0.2 ml, group 4), and CIK cells combined with MMC (group 5), respectively.

RESULTSThe percentage of CD3(+), CD3(+)CD8(+), CD3(+)CD56(+), CD25(+) cells increased from 64.0%, 28.0%, 7.8%, and 9.1% to 94.7%, 67.7%, 61.3%, and 84.0% respectively after cytokine induction. The percentage of CD3(+) and CD3(+)CD8(+) cells remained at high levels during incubation period, but that of CD25(+) and CD3(+)CD56(+) cells peaked respectively on day 7 and 13 and then declined. During the 90-day observation, the tumor formation rates were 100%, 70.0%, 80.0%, 70.0% and 66.7%; and the mouse survival rates were 10.0%, 60.0%, 40.0%, 50.0% and 75.0%, respectively from group 1 to group 5. Compared to the other groups, in the combined therapy group of mice, not only the tumor grew slowly and but also showed more marked tissue necrosis.

CONCLUSIONThe growth inhibitory effect on human HCC transplanted in nude mice of combined CIK cells and MMC treatment is more potent than that of CIK cells or MMC alone.

Animals ; Antibiotics, Antineoplastic ; therapeutic use ; Carcinoma, Hepatocellular ; immunology ; pathology ; therapy ; Cell Line, Tumor ; Cells, Cultured ; Combined Modality Therapy ; Cytokines ; metabolism ; pharmacology ; Female ; Humans ; Immunotherapy, Adoptive ; Killer Cells, Natural ; transplantation ; Liver Neoplasms ; immunology ; pathology ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mitomycin ; therapeutic use ; Neoplasm Transplantation

AIMThe purpose of this study was to develop a mathematical model to quantitatively describe the passive transport of macromolecules within dental biofilms.

METHODOLOGYFluorescently labeled dextrans with different molecular mass (3 kD, 10 kD, 40 kD, 70 kD, 2000 kD) were used as a series of diffusion probes. Streptococcus mutans, Streptococcus sanguinis, Actinomyces naeslundii and Fusobacterium nucleatum were used as inocula for biofilm formation. The diffusion processes of different probes through the in vitro biofilm were recorded with a confocal laser microscope.

RESULTSMathematical function of biofilm penetration was constructed on the basis of the inverse problem method. Based on this function, not only the relationship between average concentration of steady-state and molecule weights can be analyzed, but also that between penetrative time and molecule weights.

CONCLUSIONThis can be used to predict the effective concentration and the penetrative time of anti-biofilm medicines that can diffuse through oral biofilm. Furthermore, an improved model for large molecule is proposed by considering the exchange time at the upper boundary of the dental biofilm.

Actinomyces ; growth & development ; Algorithms ; Biofilms ; growth & development ; Biological Transport ; Dental Plaque ; microbiology ; Dextrans ; pharmacokinetics ; Diffusion ; Fluorescent Dyes ; pharmacokinetics ; Fusobacterium nucleatum ; growth & development ; Macromolecular Substances ; pharmacokinetics ; Microscopy, Confocal ; Models, Biological ; Molecular Probe Techniques ; Streptococcus mutans ; growth & development ; Streptococcus sanguis ; growth & development

BACKGROUND: Oxidative stress is one of the important mechanisms of postoperative abdominal adhesion. The nuclear factor-E2-related factor 2 (Nrf2)-antioxidant response element (ARE) signal pathway is an important endogenous anti-oxidation stress pathway. Our previous study found that ligustrazine nano-spray can inhibit the formation of postoperative abdominal adhesion in rats, and moreover, ligustrazine has an anti-oxidation effect. OBJECTIVE: To observe the effect of ligustrazine nano-spray on the expression of mRNAs and proteins related to the Nrf2-ARE signal pathway in rats with abdominal adhesion, and to investigate the mechanism by which ligustrazine nano-spray inhibits abdominal adhesion via regulating the Nrf2-ARE signal pathway. METHODS: Sprague-Dawley rats were randomly divided into sham group, model group, ligustrazine group and sodium hyaluronate group. In the sham group, only laparotomy was performed without modeling. In the model group, an abdominal adhesion model was created but no drug was used. In the ligustrazine group, ligustrazine nano-spray was used on the wound before incision suturing. In the sodium hyaluronate group, sodium hyaluronate was applied on the wound before incision suturing. RESULTS AND CONCLUSION: Compared with the model group, ligustrazine nano-spray reduced the levels of reactive oxygen species, tissue inhibitor of matrix metalloproteinase 1, but increased the level of matrix metalloproteinase 9 in the rat serum. The expression of Nrf2 and HO-1 mRNA and proteins was also up-regulated in the ligustrazine group relative to the model group. Therefore, ligustrazine nano-sprays can inhibit abdominal adhesions in rats, and its mechanism may be related to the regulation of Nrf2 and heme oxygenase 1 mRNA and proteins expression and the activation of Nrf2-ARE signaling pathway.

BACKGROUND:Internal and external fixation combined with autologous bone graft for treating atrophic nonunion has a long treatment cycle,and moreover,it cannot achieve a 100％ cure rate.Platelet-rich plasma contains a variety of growth factors and a large number of white blood cells,and contributes to tissue healing.However,there is no clinical study on the effectiveness of platelet-rich plasma combined with conventional surgery in the treatment of atrophic nonunion.OBJECTIVE:To investigate the effectiveness of platelet-rich plasma in the treatment of atrophic nonunion of femoral shaft fractures.METHODS:We conducted a prospective,open-label,randomized,controlled clinical trial at the Affiliated Hospital of Qinghai University,China.Ninety-two patients with atrophic nonunion of femoral shaft fractures were equally and randomly divided into control group and experimental group.Patients in the control group received conventional surgery.Patients in the experimental group were injected with autologous platelet-rich plasma on the basis of conventional surgery.The primary outcome was fracture healing rate at postoperative 9 months.The secondary outcomes were visual analogue scale scores in resting state and during passive motion,healing time,treatment costs,and adverse reactions.The study protocol was approved by the Ethics Committee of Affiliated Hospital of Qinghai University of China (approval number:QHG0223A) on May 20,2014.Written informed consent was provided by each patient and their family members after they fully understood the treatment plan.RESULTS AND CONCLUSION:Our partial results demonstrated that visual analogue scale scores and complications were similar between the two groups at postoperative 1-3 days.The healing rate was significantly higher in the experimental group than in the control group.The healing time was significantly shorter in the experimental group than in the control group.This trial will provide objective data for the clinical use of platelet-rich plasma combined with conventional surgery for the treatment of atrophic nonunion.

OBJECTIVETo investigate the effects of lentivirus mediated siRNA targeting human metastasis suppressor 1 (MTSS1, MIM-B gene) gene on the invasive and metastatic potentials of hepatocellular carcinoma (HCC) MHCC97H cells.

METHODSThe siRNA targeting MTSS1 was cloned into one lentivirus work vector. The work vector and three package plasmids were co-transfected into 293T cells with the help of lipefeetamine 2000. Lentivirus was collected in 72 hours and was added to the cultured MHCC97H cells. The total cell MIM-B mRNA and MIM-B protein were extracted and underwent real-time PCR and western-blot test respectively. Boden chamber assay was used to evaluate the invasive potential of MHCC97H cells. Gelatin zymography was used to detect matrix metalloproteinase-2 (MMP2) activity. Metastatic human HCC nude mice models were established by orthotopic implantation with a high metastatic potential human HCC cell line MHCC97H. Twenty-four nude mice bearing orthotopic xenografts were randomized into black control group, Lenti-GFP group and intervention group (Lenti-MTSS1 group) 14 days after orthotopic implantation (8 per group). The ultrasound-guided multi-point injection was performed on mice with borate buffered saline, Lenti-GFP and Lenti-MTSS1 respectively. Mice were sacrificed on day 35 for the examination of pulmonary metastasis. The SPSS 13.0 soft ware was applied to data analysis.

RESULTSThe small interfering RNA targeting MTSS1 was constructed successfully with a transfection efficiency of 97.0%, which produced a marked inhibition of invasive ability of MHCC97H cells through Matrigel, being 37.9+/-4.4, 37.4+/-5.3 and 26.6+/-4.6 in the black control group, Lenti-GFP group and Lenti-MTSS1 group (F = 26.695, P value is less than 0.01), respectively. MIM-B expression and MMP2 activity of intervention group were also significantly down-regulated as compared to the control group. The results of in vivo studies showed that the numbers of lung metastatic nodules were 6.5+/-2.6, 6.4+/-2.7 and 3.8+/-1.3 in the black control group, Lenti-GFP group and intervention group respectively with significant statistical difference (F = 3.637, P value is less than 0.05), accorded with tumor tissue MIM-B mRNA expression of 0.39+/-0.19, 0.38+/-0.10 and 0.16+/-0.11 respectively (F = 11.644, P value is less than 0.01) when comparison was made between control group and therapy group.

CONCLUSIONSmall interfering RNA mediated by lentivirus inhibited MIM-B expression and resulted in inhibition of the invasive and metastatic potentials of MHCC97H cells, which may attributed, in part, the down regulation of MMP2 activity, and thus may provide a new molecular targeted therapy for HCC patients in the future.

Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microfilament Proteins ; genetics ; Neoplasm Proteins ; genetics ; Neoplasm Transplantation ; RNA, Small Interfering ; genetics ; Transfection

BACKGROUNDSeveral studies using functional magnetic resonance imaging (fMRI) and positron emission tomography (PET) have indicated that cognitive remediation therapy (CRT) might improve cognitive function by changing brain activations in patients with schizophrenia. However, the results were not consistent in these changed brain areas in different studies. The present activation likelihood estimation (ALE) meta-analysis was conducted to investigate whether cognitive function change was accompanied by the brain activation changes, and where the main areas most related to these changes were in schizophrenia patients after CRT. Analyses of whole-brain studies and whole-brain + region of interest (ROI) studies were compared to explore the effect of the different methodologies on the results.

METHODSA computerized systematic search was conducted to collect fMRI and PET studies on brain activation changes in schizophrenia patients from pre- to post-CRT. Nine studies using fMRI techniques were included in the meta-analysis. Ginger ALE 2.3.1 was used to perform meta-analysis across these imaging studies.

RESULTSThe main areas with increased brain activation were in frontal and parietal lobe, including left medial frontal gyrus, left inferior frontal gyrus, right middle frontal gyrus, right postcentral gyrus, and inferior parietal lobule in patients after CRT, yet no decreased brain activation was found. Although similar increased activation brain areas were identified in ALE with or without ROI studies, analysis including ROI studies had a higher ALE value.

CONCLUSIONSThe current findings suggest that CRT might improve the cognition of schizophrenia patients by increasing activations of the frontal and parietal lobe. In addition, it might provide more evidence to confirm results by including ROI studies in ALE meta-analysis.

Brain ; physiopathology ; Cognition ; Cognitive Remediation ; Humans ; Likelihood Functions ; Magnetic Resonance Imaging ; Positron-Emission Tomography ; Schizophrenia ; diagnostic imaging ; therapy