OBJECTIVEThis study aims to evaluate the effect of ptxA and ptxB genes, which are important genes in the L-ascorbate phosphotransferase system (PTS) of Streptococcus mutans (S. mutans).

METHODSThe ptxA-, ptxB-, and ptxAB-double deficient mutant as well as ptxAB-complemented strain were constructed. Quantitative real-time polymerase chain reaction analysis was performed to evaluate the expression of the target genes of wild-type S. mutans when L-ascorbate was used as the sole carbohydrate source. The OD₆₀₀ values of the wild type, deficient, and complemented strains were continuously monitored, and their growth curves were constructed to compare growth capacity.

RESULTSPolymerase chain reaction and sequencing analyses suggested that deficient and complemented strains were successfully constructed. The expression levelsof ptxA and ptxB significantly increased (P < 0.01) when L-ascorbate was used as the sole carbohydrate source. The growth capacity of the deficient mutants decreased compared with that of the wild-type strain. However, the wild-type phenotype could be restored in the complemented strain.

CONCLUSIONptxA and ptxB genes are associated with L-ascorbate metabolism of S. mutans. The construction of deficient strains and complemented strain lay a foundation for further mechanism study on L-ascorbate metabolism in S. mutans.

Bacterial Proteins ; genetics ; Genes, Bacterial ; Phosphotransferases ; metabolism ; Real-Time Polymerase Chain Reaction ; Streptococcus mutans ; physiology ; Transcription Factors ; genetics

The aim of this study is to investigate the anti-inflammatory effect of the adenosine derivative N6-(3-hydroxylaniline) adenosine (WS070117M1) on cigarette smoke plus LPS (lipopolysaccharide)-induced chronic obstructive pulmonary disease (COPD) in mice and its mechanism. COPD model was established by exposing male BALB/c mice to cigarette smoke and challenged with LPS inhalation. Supernatants of bronchoalveolar lavage fluid (BALF) were harvested and IL-1β, IL-6, IL-8 and TGF-β1 levels were measured by ELISA (enzyme-linked immunesorbent assay). The number of total white blood cells and neutrophils in bronchoalveolar lavage fluid was counted separately. Lung tissue was stained with Mayer 's hematoxylin and eosin for histopathologic examination. pAMPKa protein expression and distribution of lung tissue were analyzed by immunohistochemistry method. In vitro, levels of AMPKα phosphorylation in phorbol-12- myristate-13-acetate (PMA) differentiated THP-1 cells was detected by immunohistochemistry, IL-8 level in supernatants of cigarette smoke condensate stimulating PMA differentiated THP-1 cells was measured by ELISA. The results showed that WS070117M1 treatment significantly activated AMPKa in the lung tissue. It also resulted in down regulation of IL-1β, IL-6, IL-8 and TGF-β1 levels in bronchoalveolar lavage fluid and IL-8 level in cigarette smoke condensate stimulating PMA differentiated THP-1 cells. In addition, WS070117M1 could inhibit the recruitment of total white blood cells and neutrophils. These results suggest that WS070117M1 may alleviate the airway inflammation by activating AMPK in the lung tissue.
AMP-Activated Protein Kinases ; metabolism ; Adenosine ; analogs & derivatives ; Animals ; Bronchoalveolar Lavage Fluid ; Cell Line, Tumor ; Disease Models, Animal ; Humans ; Inflammation ; drug therapy ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocyte Count ; Lipopolysaccharides ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophils ; cytology ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Smoke ; adverse effects ; Tobacco ; Transforming Growth Factor beta1 ; metabolism

Both market research and literature reports both found that the ootheca of mantodea was all used as medicine. However, Chinese Pharmacopoeia only records the ootheca of three mantis species. The clinical use of ootheca unrecorded in Chinese Pharmacopoeia, will pose potential risks to drug safety. It's urgent to identify the origin of Mantidis Oötheca. The current researches about original animal in Mantidis Oötheca are based on morphology and unanimous. DNA barcoding fill gaps of the traditional morphological identification, which is widely used in animal classification studies. This study first use DNA barcoding to analyze genetic distance among different Mantidis Oötheca types, align COI sequences between mantis and Mantidis Oötheca and construct the phylogeny tree. The result confirmed that Tenodera sinensis and Hierodula patellifera were the origin insects of Tuanpiaoxiao and Heipiaoxiao, respectively, and Statilia maculate and Mantis religiosa were the origin insects of Changpiaoxiao.
Animals ; DNA ; genetics ; DNA Barcoding, Taxonomic ; methods ; Electron Transport Complex IV ; genetics ; Insect Proteins ; genetics ; Mantodea ; classification ; genetics ; Medicine, Chinese Traditional ; Phylogeny

OBJECTIVETo assess the clinical therapeutic effects of elastic intramedullary nail on extremity fractures in children.

METHODSFrom June 2005 to March 2008, 40 children with extremity fractures were treated by elastic intramedullary nail, in whom femoral shaft fractures occurred in 26 cases, tibiofibular fractures in 8 cases, radial capitular fractures in 4 cases, ulnoradial fractures in 2 cases. All patients were treated by closed reduction and elastic intramedullary nail fixation.

RESULTSAll the fractures gained satisfactory reduction and healing. The average duration needed for fracture healing was 1-2 months. Postoperative follow-up confirmed a sound functional recovery.

CONCLUSIONSThe elastic intramedullary nail is a minimally invasive and effective surgical approach for treatment of extremity fractures in children. It allows early functional exercises after operation and secures a satisfactory bone union and functional recovery.

Adolescent ; Bone Nails ; Child ; Child, Preschool ; Extremities ; diagnostic imaging ; injuries ; surgery ; Female ; Fracture Fixation, Intramedullary ; instrumentation ; Fracture Healing ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Male ; Minimally Invasive Surgical Procedures ; Postoperative Complications ; Radiography ; Treatment Outcome

The aim of the present study is to investigate the effects of Vam3 which is one of the dihydroxystilbene compounds on expressions of ICAM-1 in the lungs of OVA-induced asthmatic mice and the mechanisms of anti-airway inflammation. Balb/c mice were challenged with OVA inhalation. Lung tissues were stained with Mayer's hematoxylin and eosin for histopathologic examination. The expression of ICAM-1 in the lungs of mice was analyzed by Western blotting and immunohistochemistry method. The NF-kappaB activities were detected by NF-kappaB-luc reporter genetic transient transfection method. The activities of MMP-9 induced by LPS, TNF-alpha and PMA in THP-1 cells were determined by gelatin zymography method. The results showed that Vam3 could inhibit the expression of ICAM-1 in the OVA-induced mouse model. In addition, Vam3 could significantly suppress the activities of NF-kappaB in A549 cells and MMP-9 in THP-1 cells induced by LPS, TNF-alpha and PMA. These results suggested that Vam3 could alleviate the asthmatic inflammation by decreasing ICAM-1 expression in asthmatic mice, down regulating NF-kappaB and MMP-9 activities. Compound Vam3 showed inhibitory effects on inflammatory signal pathways involved in asthma.
Animals ; Anti-Asthmatic Agents ; pharmacology ; Anti-Inflammatory Agents ; pharmacology ; Asthma ; chemically induced ; metabolism ; Benzofurans ; pharmacology ; Cell Line, Tumor ; Humans ; Inflammation ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Leukemia, Myeloid ; metabolism ; pathology ; Lung ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; metabolism ; Ovalbumin ; Stilbenes ; pharmacology

OBJECTIVETo analyze the characteristics of CDR3 of TCRbeta on CD8+ T cells in chronic hepatitis B patients.

METHODSEight patients with chronic hepatitis B (ALT more than 2 ULN) were enrolled in this study. CD8+ T cells were isolated from peripheral blood. RT-PCR was proformed to amplify the CDR3 of TCRbeta, and the PCR products were sequenced and analyzed.

RESULTSThe chronic hepatitis B patients showed obvious clonal expansion of T cell, and three perturbation patterns of T cell expansion were showed in the CDR3 of TCRbeta, including monoclonicity, oligoclonicity and skewed peak patterns. The number of perturbation families of CD8+ subpopulation was significantly higher than that of CD8- subpopulation (10.6+/-4.7 vs. 4.1+/-3.1, t = 6.619, P less than 0.01). In 3 out of 8 patients, the number of perturbation families of CD8+ subpopulation was also higher than that of PBMCs without depleting CD8+ subpopulation.

CONCLUSIONSThe characteristics of CDR3 of TCRbeta may help to understand the inflammatory response in CHB patients.

Adult ; CD8-Positive T-Lymphocytes ; immunology ; Complementarity Determining Regions ; genetics ; Genes, T-Cell Receptor beta ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Male ; Receptors, Antigen, T-Cell ; immunology ; Young Adult

Biotinylated chitosan nanoparticles (Bio-CS-NP) were prepared for the active delivery to cancer cells and its characterization was investigated in this study. The preparation process included two steps. First, biotinylated chitosan ( Bio-CS ) was obtained through a reaction between sulfosuccinimidobiotin and chitosan (CS). Second, Bio-CS-NP were prepared by the precipitation of Bio-CS with sodium chloride solution. With a biotin reagent box, the conjugation densities of biotin on the surface of Bio-CS-NP were determined. The morphology and diameter of the nanoparticles were assayed by transmission electron microscope (TEM) and laser light scattering particle analyzer, respectively. The uptake of nanoparticles by human hepotacarcinoma HepG2 cells, for example, Bio-CS-NP and chitosan nanoparticles (CS-NP) without any modification, was quantitatively examined. The results indicated that the conjugation densities of biotin on the surface of Bio-CS-NP were 2.2 biotin CS. Bio-CS-NP were spherical, smooth on the surface. The average diameter was 296.8 nm. The polydispersion index was 0.155. The uptake of Bio-CS-NP by HepG2 cells was much higher than that of CS-NP (P < 0.05). It demonstrated that Bio-CS-NP can be applied as a new vehicle to actively deliver anticancer drugs to tumor cells. The method for the determination of biotin was simple and practical.
Biotin ; analogs & derivatives ; chemistry ; metabolism ; Biotinylation ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Chitosan ; chemistry ; metabolism ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Nanoparticles ; Particle Size ; Succinimides ; chemistry ; metabolism

OBJECTIVETo search for hepatocellular carcinoma (HCC) invasion related biomarkers using the cell membrane proteomics approaches, and to validate the markers using experimental and clinical specimens.

METHODSThe HCCLM9 and MHCC97L cells with a similar genetic background and remarkably different metastasis behaviors were used for comparative membrane proteome profiling using sodium dodecyl sulfate polyacrylamide gel electrophoresis and electrospray ionization mass spectrometry technologies. Candidate protein makers were further validated by western blot on cells, immunohistochemistry (IHC) on animal tumor tissues, and tissue micro-array on clinical specimens.

RESULTSThe membrane proteins of MHCC97L and HCCLM9 cells were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis analyses. 14 proteins were identified by ESI-MS/MS among the differential bands. Coronin-1C was overexpressed in HCCLM9 (7.31+/-0.73) versus MHCC97L (2.84+/-0.99) validated by western blot. Elevated coronin-1C expression was observed in liver cancer tissues of HCCLM9 nude mice. IHC study in 115 human HCC specimens demonstrated that patients with higher coronin-1C expression had more advanced stage.

CONCLUSIONThe study suggests that coronin-1C could be a potential molecule to predict HCC invasive behavior.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microfilament Proteins ; biosynthesis ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis

Background and Objectives: Systemic scleroderma (SSc) is a rare and serious connective tissue disease, an autoimmune disease, and a rare refractory disease. In this study, preventive effect of single systemic human umbilical cord mesenchymal stem cells (UC-MSCs) transfusion on SSc was preliminarily explored. Methods: and Results: SSc mouse model was established by daily intradermal injection of Hypochlorite (HOCl). SSc mice were treated by single transfusion of UC-MSCs at 0.625×10 5 , 2.5×105 and 1×106 respectively. At the 42nd day of intradermal injection of HOCl, the symptoms showed up by skin and alveolar wall thickening, lymphocytic infiltration, increased collagen in skin/lung, and the increased proportion of CD3 ＋ CD4＋ CD25＋ FoxP3＋ cells (a Treg subset) in spleen. After UC-MSCs transfusion, the degree of skin thickening, alveolar wall thickening and lymphocyte infiltration were decreased, the collagen sedimentation in skin/lung was decreased, and the proportion of CD3＋ CD4＋ CD25＋FoxP3＋ cells was decreased. Conclusions UC-MSC can achieve a preventive effect in SSc mice by fibrosis attenuation and immunoregulation.

Background and Objectives: Systemic scleroderma (SSc) is a rare and serious connective tissue disease, an autoimmune disease, and a rare refractory disease. In this study, preventive effect of single systemic human umbilical cord mesenchymal stem cells (UC-MSCs) transfusion on SSc was preliminarily explored. Methods: and Results: SSc mouse model was established by daily intradermal injection of Hypochlorite (HOCl). SSc mice were treated by single transfusion of UC-MSCs at 0.625×10 5 , 2.5×105 and 1×106 respectively. At the 42nd day of intradermal injection of HOCl, the symptoms showed up by skin and alveolar wall thickening, lymphocytic infiltration, increased collagen in skin/lung, and the increased proportion of CD3 ＋ CD4＋ CD25＋ FoxP3＋ cells (a Treg subset) in spleen. After UC-MSCs transfusion, the degree of skin thickening, alveolar wall thickening and lymphocyte infiltration were decreased, the collagen sedimentation in skin/lung was decreased, and the proportion of CD3＋ CD4＋ CD25＋FoxP3＋ cells was decreased. Conclusions UC-MSC can achieve a preventive effect in SSc mice by fibrosis attenuation and immunoregulation.