This study sought to investigate the in vivo antiviral effect of amantadine (AM) and biphenyl dimethyl dicarboxylate (DDB) on hepatitis B virus (HBV) in HBV replication mice. HBV replication-competent plasmid was transferred into male BALB/c mice by using hydrodynamics-based in vivo transfection procedure to develop HBV replication mouse model. The model mice were matched by body weigh, age and serum levels of hepatitis B e antigen (HBeAg) and were divided into four groups: AM group, DDB group, AM+DDB group and NS group, with the last one as control, and the mice of each group were administered corresponding agent orally twice a day, in a medication course lasting 3 d. On the third day, the mice were sacrificed 4-6 h after the last oral intake. HBV DNA replication intermediates in liver were analyzed by Southern blot hybridization. The serum hepatitis B surface antigen (HBsAg) and HBeAg were detected by enzyme linked immunosorbent assay (ELISA). Compared to the animals in the control group, HBV DNA replication intermediates in liver and HBsAg and HBeAg in serum from the AM and AM plus DDB group of mice decreased, and there was no difference between these two groups of mice. The levels of HBV DNA intermediate from liver and the serum HBsAg and HBeAg between the control and DDB group, however, were not obviously different. In conclusion, the inhibition effect of AM on HBV was detected, but treatment with DDB for 3 days did not influence the viral replication and expression of HBV in the HBV replication mice.
Amantadine ; pharmacology ; Animals ; Antiviral Agents ; pharmacology ; DNA Replication ; DNA, Viral ; biosynthesis ; Dioxoles ; pharmacology ; Disease Models, Animal ; Hepatitis B ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; Plasmids ; Transfection ; Virus Replication ; drug effects

OBJECTIVETo evaluate the soft profile and hard tissue changes after maxillary protraction with skeletal anchorage implant in treatment of Class III malocclusion during growth period.

METHODS18 patients with skeletal Class III malocclusion were treated with maxillary protraction for about 9 months, (3.5 +/- 0.1) N, with skeletal anchorage implant and face mask. Cephalometric records were analyzed to assess the changes of maxillo-facial structure of the hard and soft tissue before and after treatment.

RESULTSAll patients' Class III profiles were corrected. Maxillary growth increased, chin clockwise rotated, facial convexity angle increased, lower lip protrusion decreased. Facial vertical height ratio, nasolabial angle, upper lip protrusion and mentolabial sulcus changed unconspicuously. Upper incisors kept in sites, lower incisor upright, maxilla moved forwards. SNA, ANB significantly increased. SNB decreased and the mandible clockwise rotated.

CONCLUSIONThe maxilla is effectively protracted without significant rotation by using skeletal anchorage implant. The undesired effects of conventional protraction therapies, such as labial tilt of upper anterior teeth and extrusion of the maxillary molars, are reduced or eliminated with skeletal anchorage implant. These effects can conspicuously correct profiles of the patients with skeletal Class III malocclusion, make the profile more harmonious and aesthetic.

Cephalometry ; Chin ; Extraoral Traction Appliances ; Face ; Humans ; Incisor ; Lip ; Malocclusion, Angle Class III ; Mandible ; Maxilla ; Molar

Objective To investigate the immunological pathogenesis of immune reconstitution inflammatory syndrome (IRIS) during highly active antiretroviral therapy( HAART), in this prospective cohort study we analyzed the lymphocyte subsets, lymphocyte activation, changes in regulatory T cells, and levels of Th1 and Th2 cytokines in both IRIS and non-IRIS groups. Methods Two hundred and thirty-eight AIDS patients received HAART and participated prospective research cohort for 24 weeks follow-up. Forty-seven IRIS cases and 191 non-IRIS cases were enrolled in the IRIS group or non-IRIS group respectively. Blood samples were collected in both groups at pre- and post-HAART 12 weeks, 24 weeks. Using flow cytometer to detect the immunophenotypes of lymphocyte subsets (CD4 + CD45RA+ CD62L+, CD8+ CD45RA+ CD62L+naive T cells; CD4+ CD45RO+, CD8+ CD45RO+ memory T cells), activated T lymphocytes (CD4+CD38 +, CD8 + CD38 + cells), and regulatory T cell ( CD4 + CD25 + Foxp3 + ). Blood samples collected at pre-and post-HAART4 weeks, 12 weeks, 24 weeks and used ELISA to detect IL-2, IFN-γ, IL-4, IL-10and IL-7 cytokine serum levels. Results The percentages of CD4 + and CD8 + naive T cells and mlemory T cells exhibited no significant differences at the baseline, 12 weeks, 24 weeks of HAART initiation between both groups, but CD4 + and CD8 + memory T cells were demonstrated a trend towards to increase while compared to baseline during HAART. The percentages of CD4 + and CD8 + activated T cells are significantly higher at the baseline while compared to normal control and demonstrated a downward trend, but between both groups showed no significant difference. The percentages of CD4 + regulatory T cell was lower in IRIS group than non-IRIS group at the baseline, 12 weeks, 24 weeks and the onset of IRIS. Th1 cytokines, IL-2 and IFN-γshowed an upward trend during HAART at the levels of IRIS group had significantly increased at 4 weeks and the onset of IRIS. Th2 cytokines, IL-4 and IL-10 showed a downward trend during HAART,and the levels of IL-10 in IRIS group had significantly decreased at 4 weeks and the onset of IRIS. IL-7 was higher than normal control at the baseline in two groups and showed a downward trend during HAART. The level of IL-7 was higher than non-IRIS group at all follow-up points. Conclusion Memory T cells appear rapid increase in the early stage of HAART and may play a significant role in the inflammatory response of IRIS. CD4 + and CD8 + naive T cells, memory T cells and activated T cells showed no significant difference between IRIS and non-IRIS group within 24 weeks after HAART started. There was a significant reduction in the frequency of regulatory T cells in IRIS group without obvious upward trend during HAART, suggesting that the immune suppression function of regulatory T cells in IRIS was impaired. IL-2 and IFN-γ significantly increased while IL-10 significantly decreased at 4 weeks post-HAART initiation and onset of IRIS in IRISgroup than non-IRIS group, suggested that IRIS was related to cytokines environment disorder. That is, a significant increase in inflammatory cytokines, while the relative lack of non-inflammatory cytokines. The level of IL-7 decreased gradually after HAART started, and it was higher in IRIS group when compared to non-IRIS group in the first 24 weeks after HAART started. Also IL-7 may play a role in the pathogenesis of IRIS.

OBJECTIVETo analyze the associated predictive factors of lymph node metastasis in intramucosal early gastric cancer, and to provide reference for individualized treatment protocol.

METHODSClinical data of 498 patients who were pathologically confirmed to be intramucosal early gastric cancer after radical resection of gastric cancer were retrospectively reviewed. Univariate and multivariate analyses were applied to evaluate the associated factors.

RESULTSForty-three out of 498 patients (8.6%) had lymph node metastasis. Tumor size (HR=1.525, 95%CI:1.040-2.236), depth of invasion (HR=1.656, 95%CI:1.158-2.368) and histological type (HR=8.149, 95%CI:1.770-37.513) were the independent risk factors for lymph node metastasis in intramucosal early gastric cancer.

CONCLUSIONSPrediction and therapeutic guidance for lymph node metastasis in intramucosal early gastric cancer can be carried out according to clinicopathological risk factors.

Adult ; Female ; Humans ; Logistic Models ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; pathology ; surgery

OBJECTIVETo demonstrate the vasodilatation activity of the coumarin-containing Angelica dahurica var. formosana and to further analyze active components in the herb extracts.

METHODS(1) The vasodilatation effects induced by different extracts (cyclohexane, ethyl acetate, acetone, methanol, 95 % ethanol and water) of Angelica dahurica var. formosana on mouse thoracic aorta pre-contracted with phenylephrine were investigated. (2) The amount of imperatorin and isoimperatorin in each extract was measured by high-performance liquid chromatography. (3) The vasodilatation effects of imperatorin and isoimperatorin on mouse thoracic aorta were compared using the same in vitro method. (4) The vasodilatation mechanism of imperatorin in the mouse thoracic aorta pre-contracted with phenylephrine was studied using the methods of denuded endothelium, NG-nitro-L-arginine methylester (L-NAME, a nitric oxide synthase inhibitor), and propranolol.

RESULTS(1) The cyclohexane and ethyl acetate extracts of Angelica dahurica var. formosana decreased the maximal response of phenylephrine-induced mouse thoracic aorta contraction dose-dependently, with 50% inhibiting concentration (IC(50)) values of 35.3+/-12.4 mg/L and 40.5+/-12.0 mg/L, respectively. The vasodilatation effect of imperatorin and isoimperatorin was dose-dependent. (2) The cyclohexane extract, showing the strongest vasodilatation effect, possessed the highest contents of imperatorin (4.09%) and isoimperatorin (0.27%, w/w). There was a correlation between the vasodilatation activity and the contents of imperatorin and isoimperatorin in the extracts. (3) The vasodilatation effect of imperatorin was about 4-fold stronger than that of isoimperatorin. (4) The vasodilatation effect of imperatorin was signifificantly attenuated to 24.88%+/-4.06% in the denuded endothelium group compared with the intact endothelium group. And 1 mmol/L L-NAME reduced the imperatorin-induced vasorelaxation by 32.18 %+/-11.29 %.

CONCLUSIONSThe principal effective component of Angelica dahurica var. Formosana was found to be imperatorin. Imperatorin-induced vasodilatation is at least partially regulated by nitric oxide, and has no correlation to beta-receptor.

Angelica ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Endothelium, Vascular ; physiology ; Furocoumarins ; analysis ; pharmacology ; Male ; Mice ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; physiology ; Phenylephrine ; pharmacology ; Plant Extracts ; pharmacology ; Propranolol ; pharmacology ; Vasodilation ; drug effects

Tooth bleaching agents may weaken the tooth structure. Therefore, it is important to minimize any risks of tooth hard tissue damage caused by bleaching agents. The aim of this study was to evaluate the effects of applying 45S5 bioglass (BG) before, after, and during 35% hydrogen peroxide (HP) bleaching on whitening efficacy, physicochemical properties and microstructures of bovine enamel. Seventy-two bovine enamel blocks were prepared and randomly divided into six groups: distilled deionized water (DDW), BG, HP, BG before HP, BG after HP and BG during HP. Colorimetric and microhardness tests were performed before and after the treatment procedure. Representative specimens from each group were selected for morphology investigation after the final tests. A significant color change was observed in group HP, BG before HP, BG after HP and BG during HP. The microhardness loss was in the following order: group HP>BG before HP, BG after HP>BG during HP>DDW, BG. The most obvious morphological alteration of was observed on enamel surfaces in group HP, and a slight morphological alteration was also detected in group BG before HP and BG after HP. Our findings suggest that the combination use of BG and HP could not impede the tooth whitening efficacy. Using BG during HP brought better protective effect than pre/post-bleaching use of BG, as it could more effectively reduce the mineral loss as well as retain the surface integrity of enamel. BG may serve as a promising biomimetic adjunct for bleaching therapy to prevent/restore the enamel damage induced by bleaching agents.
Animals ; Biomimetic Materials ; analysis ; therapeutic use ; Cattle ; Ceramics ; analysis ; chemistry ; Chemical Phenomena ; Color ; Colorimetry ; Dental Enamel ; drug effects ; ultrastructure ; Electron Probe Microanalysis ; Glass ; analysis ; chemistry ; Hardness ; Hydrogen Peroxide ; pharmacology ; Hydrogen-Ion Concentration ; Microscopy, Electron, Scanning ; Protective Agents ; analysis ; therapeutic use ; Random Allocation ; Solubility ; Spectroscopy, Fourier Transform Infrared ; Time Factors ; Tooth Bleaching ; methods ; Tooth Bleaching Agents ; pharmacology ; Water ; chemistry ; X-Ray Diffraction

Human bone marrow is the major source of mesenchymal stem cells (MSC). It was reported that the standard density gradient centrifugation method was not efficient in isolating MSC and it may be caused by the existing of bone marrow particles. In previous study, a lot of MSC were obtained by culturing bone marrow particles alone combined with standard method. However, it is time- and labor-consuming to obtain bone marrow particles by filtering and to isolate MNC by density gradient centrifugation. This study was purposed to explore the more simple and efficient method to isolate MSC from bone marrow. Seven normal bone marrow aspirates were collected and centrifugated. The bone marrow particles floated on surface layers were cultured by modified primary explant culture, whereas the bone marrow aspirates deposited were cultured by direct plating method, then the immun phenotype and differentiation capability of isolated cells were analyzed. The results showed that in 3 of 7 aspirates, bone marrow particles were floated on surface layers, whereas the other bone marrow cells and some particles were deposited after centrifugation. The MSC were reliably isolated from the floating layers or deposited aspirates by modified primary explant culture and direct plating method separately. After 3 passages the isolated MSC did not express CD45 and CD34, but expressed CD105, CD73, CD44, CD90, CD49e and they could differentiate into chondrocytes and adipocytes. It is concluded that normal human bone marrow MSC can be isolated simply and efficiently by direct plating method in combination with modified primary explant culture.
Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology

BACKGROUNDCancer testis antigens (CTAs) are a novel group of tumor associated antigens. Demethylating agent decitabine was reported to be able to up-regulate CTAs through its hypomethylation mechanism, thus enhance the immunogenicity of leukemia cells. However, few researches have ever focused on the questions that whether this immunostimulatory effect of decitabine could induce autologous CTA specific cytotoxic T lymphocytes (CTLs) in vivo, and if so, whether this effect contributes to disease control. In this study, we aimed to show that decitabine could induce specific autologous CTLs against some mouse CTAs in leukemia cells in vitro and in vivo.

METHODSSeveral mouse CTAs were screened by RT-PCR. CTL specific to one of the CTAs named P1A was detected and sorted by P1A specific dimer by flow cytometry. The activity of specific CTLs was measured by real time RT-PCR.

RESULTSWe firstly screened expression of some CTAs in mouse leukemia cells before and after decitabine treatment and found that decitabine treatment did up-regulate expression of many CTAs. Then we measured the CTLs' activity specific to a mouse CTA P1A in vivo and showed that this activity increased after decitabine treatment. Finally, we sorted these in vivo induced P1A specific CTLs by flow cytometry and demonstrated their cytotoxicity against decitabine treated leukemia cells.

CONCLUSIONSOur study showed the autologous immune response induced by decitabine in vivo. And more importantly, we firstly proved that this response may contribute to disease control. We believe that this immunostimulatory effect is another anti-cancer mechanism of decitabine, and this special effect would inspire new applications of decitabine in the field of leukemia treatment in the future.

Animals ; Antigens, Neoplasm ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes, Cytotoxic ; drug effects ; metabolism

Mucopolysaccharidosis type I (MPS-I) is an inborn error of metabolism with progressive multisystem involvement. Hurler syndrome is the most severe form of MPS-I that causes progressive deterioration of the central nervous system with ensuing death. This study reported the therapeutic effect of allogeneic hematopoietic stem cell transplantation (allo-HSCT) on Hurler syndrome in one case. The patient was a 25-month-old boy. He underwent allo-HSCT. The donor was his elder sister whose HLA-B locus was not matching. The reduced-intensity of BuCy conditioning regimen in allo-HSCT for this patient was as follows: busulfan 3.7 mg/kg daily at 9 to 6 days before transplantation, cyclophosphamide 42.8 mg/kg daily at 5 to 2 days before transplantation, and rabbit antithymocyte globulin 3.5 mg/kg daily at 1, 3, 5, and 7 days before transplantation. Human granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood stem cells (CD34+ cells 12.8 x10(6)/kg) were infused and cyclosporine (CSA), short-course methotrexate, daclizumab and mycophenolate mofetil (MMF) were administered to prevent graft-versus-host disease (GVHD). Complete donor-type engraftment was confirmed by Short Tandem Repeat-Polymerase Chain Reaction (STR-PCR) on day 14 after transplantation. Neutrophil and platelet engraftment occurred on days 11 and 19 after transplantation respectively. Only grade I regimen-related toxicity of live and gastrointestinal tract occurred. GVHD and graft failure were not observed. After transplantation, the clinical symptoms and the neurocognitive function were greatly improved in this patient. It was concluded that allo-HSCT was effective for the treatment of MPS-I. The reduced-intensity conditioning regimen was helpful to decrease the regimen-related toxicity. Sufficient immunosuppressive therapy and adequate hematopoietic stem cells infusion may be beneficial to the donor cell engraftment and reducing the incidence of graft failure and GVHD.
Child, Preschool ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Mucopolysaccharidosis I ; therapy ; Transplantation, Homologous

OBJECTIVETo evaluate the effect of serum luteinizing hormone (LH) variation on clinical outcomes in patients with polycystic ovarian syndrome (PCOS) undergoing in vitro fertilization-embryo transfer (IVF-ET).

METHODSA retrospective analysis was conducted in 314 patients with PCOS undergoing their first IVF cycle using standard long protocol. On the basis of LH concentrations on early-, mid- and late-follicular phase, the patients were divided into decreased LH (LH ratio≤1, group A) and increased LH (group B, LH ratio>1) groups in early- to mid-follicular phase, decreased LH (group C) and increased LH (group D) groups in mid- to late-follicular phase, and decreased LH (group E) and increased LH (group F, LH ratio>1) in early- to late-follicular phase. The clinical outcomes were compared between groups A and B, groups C and D, and between groups E and F.

RESULTSNo significant differences were found in the clinical outcomes between group A and B (P>0.05). The number of oocytes retrieved and the early abortion rate were significantly lower, but the normal fertilization rate, implantation rate, clinical pregnancy rate and ongoing pregnancy rate were significantly higher in group D than in group C (P<0.05). In group F, the early abortion rate was significantly lower and the ongoing pregnancy rate was significantly higher than those in group E (P<0.05), and no significant differences were found in other clinical outcomes between the two groups (P>0.05).

CONCLUSIONAn increase in LH level from early- or mid- to late-follicular phase has a positive effect on the clinical outcomes, but this LH variation in early- to mid-follicular phase does not affect the clinical outcomes.

Embryo Transfer ; methods ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; therapy ; Luteinizing Hormone ; blood ; Oocytes ; Polycystic Ovary Syndrome ; blood ; Pregnancy ; Pregnancy Rate ; Retrospective Studies