Adolescent ; Blood Cell Count ; Child ; Copper ; blood ; Electromagnetic Fields ; adverse effects ; Female ; Humans ; Iron ; blood ; Magnesium ; blood ; Male ; Students ; Trace Elements ; blood ; Zinc ; blood

OBJECTIVETo explore the functions of phospholipase C (PLC)-independent protein kinase C signaling pathway (PTH/nonPLC/PKC) of parathyroid hormone (PTH) and its role in bone metabolism.

METHODSOsteoblasts isolated from the calvaria of 2- or 3-day-old C57BL mice, identified by alkaline phosphatase staining and Alizarin red staining, were treated for 4 h with 100 nmol/L [Gly(1), Arg(19)]hPTH(1-28) plus 10 nmol/L RP-cAMP, 10 nmol/L [Gly(1), Arg(19)]hPTH(1-34) plus 10 nmol/L RP-cAMP , 10 nmol/L PTH(1-34), or and 0.1% trifluoroacetic acid (TFA). The total RNA was then isolated for screening differentially expressed genes related to PTH/nonPLC/PKC pathway using Affymetrix mouse 12x135K gene expression profile microarray, and the identified genes were confirmed by real-time quantitative PCR. MC3T3-E1 cells treated with [Gly(1), Arg(19)]hPTH(1-28)+RP-cAMP, [Gly(1), Arg(19)]hPTH(1-34)+RP-cAMP, [Gly(1), Arg(19)]hPTH(1-34)+ RP-cAMP +100 nmol/L Go6983, or 0.1% TFA were also examined for GR(1-28)- or GR(1-34)-mediated gene expression changes using real-time quantitative PCR.

RESULTSAlizarin red staining visualized red mineralized nodules in the osteoblasts at 28 days of culture. According to the genechip results, we selected 56 target genes related to PTH/nonPLC/PKC pathway, among which CITED1 showed higher expressions in [Gly(1), Arg(19)]hPTH(1-34)+ RP-cAMP group than in both the control group and [Gly(1), Arg(19)]hPTH(1-28)+RP-cAMP group (P<0.05), and its expression was the highest in PTH(1-34) group (P<0.05). RT-PCR of MC3T3-E1 cells yielded consist results with those in the primary osteoblasts, and the cells treated with Go6983 (a PKC inhibitor) did not show GR(1-28)- or GR(1-34)-mediated differential expression of CITED1.

CONCLUSIONThe activation of PLC-independent protein kinase C signaling pathway of PTH enhances the expression of CITED1 in mouse osteoblasts to mediate the effect of PTH on bone metabolism, and this pathway is not dependent on the activation of PLC or PKA signaling.

Animals ; Cells, Cultured ; Indoles ; Maleimides ; Mice ; Mice, Inbred C57BL ; Nuclear Proteins ; physiology ; Osteoblasts ; physiology ; Parathyroid Hormone ; physiology ; Protein Kinase C ; physiology ; Signal Transduction ; Skull ; Trans-Activators ; physiology ; Type C Phospholipases

OBJECTIVETo investigate the effect of acupoint injection of oxymatrine (OM) on experimental hepatocellular carcinoma and the mechanism.

METHODSThe rats of hepatocellular carcinoma induced by 2-acetoaminoflurence (2-AAF) were randomly divided into a normal control group (group N), a model group (group M), a control group of oxymatrine intraperitoneal injection (OM ip group) and a treatment group of small dose oxymatrine injection into Zusanli (OM ZSL group). At the end of 12h week, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (gamma-GT) were determined. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expressions of cyclin D1 and cyclin-dependent kinase 4 (CDK4) mRNA in hepatocellular carcinoma tissues.

RESULTSThe number of cancer nodes on the surface of liver in th Om ip group and the Om ZSL group was lower than in the group M, with the serum ALT, AST, and gamma-GT levels significantly decreased (P<0. 01), and significantly inhibited expressions of cyclin D1, CDK4 mRNA (P<0. 01).

CONCLUSIONOM ip and small dose oxymatrine injection into ZSL can treat or delay hepatocarcinogenisis of hepatocellular carcinoma induced by 2-AAF. Partial mechanism of this anti-carcinoma is protecting hepatocytes possibly through improving hepatic functions, and inhibiting excessive proliferation of liver cancer cells via inhibiting the expressions of cyclin Dl, CDK4 mRNA.

Acupuncture Points ; Alanine Transaminase ; blood ; Alkaloids ; administration & dosage ; Animals ; Aspartate Aminotransferases ; blood ; Cyclin D1 ; genetics ; Cyclin-Dependent Kinase 4 ; genetics ; Injections ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Quinolizines ; administration & dosage ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; gamma-Glutamyltransferase ; blood

OBJECTIVETo investigate the morphological changes after chemically extracted acellular nerve allografts transplant.

METHODSSeventy-two rabbits were divided into four groups. Acellular allografts of facial nerve were used in experimental group, and facial nerve autografts, acellular peroneal nerve allografts and peroneal nerve autografts respectively used in three control groups. The morphological changes after transplant were evaluated by modified trichrome staining, immunohistological staining and transmission electron microscope.

RESULTSThe two facial nerve grafts showed numerous regenerated nerve fibers, vessels and as well as a spindle schwann cells arranged longitudinally. No significant difference was observed in the fiber number and myelin thickness between the two groups,while the two peroneal nerve groups showed poor regeneration 6 months after operation.

CONCLUSIONSThe facial nerve allografts showed more neurite regeneration six months after transplant, and the regenerated axons passed through the distal stoma and there were well revascularized and proliferated schwann cells in the grafts.

Animals ; Disease Models, Animal ; Facial Nerve ; pathology ; transplantation ; Facial Nerve Injuries ; pathology ; surgery ; Nerve Regeneration ; Rabbits ; Transplantation, Homologous

Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.

China ; epidemiology ; Humans ; Incidence ; Pneumoconiosis ; epidemiology

OBJECTIVETo evaluate the effect of SMU.2055 gene on acid resistance of Streptococcus mutans.

METHODSA SMU.2055-dificient mutant strain of S. mutans was constructed using homologous recombination technique. The growth of the wild-type and mutant strains was monitored in both normal and acidic conditions. The lethal pH level, glycolysis, proton permeability, cell permeability and biofilm formation of the two strains were compared.

RESULTSPCR and sequence analyses verified the successful construction of the SMU.2055-dificient mutant strain. The growth and biofilm formation capacity of the mutant strain were obviously lowered in both normal and acidic conditions. The mutant strain also showed increased lethal pH level, proton permeability, and cell permeability with impaired H-ATPase activity in acidic conditions, but its minimum glycolytic pH remained unaffected.

CONCLUSIONThe SMU.2055-deficient S. mutans mutant exhibits a lowered acid resistance, which affects the growth, lethal pH, proton permeability, H-ATPase activity, cell permeability and biofilm formation but not the minimum glycolytic pH of the mutant strain.

OBJECTIVETo evaluate safety and efficacy of conversion of calcineurin inhibitors (CNI) to sirolimus (SRL) therapy for treatment of new-onset diabetes after kidney transplantation (NODAT).

METHODSOf 321 kidney transplant recipients, 34 patients who developed NODAT (10.59%) were divided into 3 groups to receive continued CNI therapy at a reduced dose (group A, 14 cases), sirolimus conversion therapy (group B, 12 cases), or oral hypoglycemic drugs (group C, 12 cases). All the patients had dietary and exercise therapies, and insulin injections were given in patients with postprandial (2 h) blood glucose over 14.0 mmol/L. The patients were followed up regularly for 5 years.

RESULTSThe mean blood glucose level was 13.02∓1.74 mol/L upon the diagnosis of NODAT in the 34 patients without significant differences between the 3 groups. At 6 months of therapy, fasting plasma glucose levels in the 3 groups decreased to 8.05 ∓2.45, 7.45∓2.44, and 9.30∓3.89 mmol/L, repsectively; at 12 months, blood glucose became normal in both groups A and B, but the patients in group A needed a greater daily insulin dose (P<0.05). In group B, the mean serum creatinine level was 165.1∓61.82 mmol/L at the conversion and lowered to 150∓53.05 mmol/L at 5 years (P<0.05), which were similar to those in group A at the two time points (152∓43.05 and 145.88∓53.05 mmol/L, respectively; P>0.05). In group C, creatinine level further increased after medication with oral hypoglycemic drugs. At 5 years, the patient and graft survival rates were 100% and 75% in group A, respectively, similar to those in group B (83.4% and 68%, respectively; P>0.05); group C showed lower patient and graft survival rates than groups B and C.

CONCLUSIONConversion from CNI to SLR therapy can significantly the metabolism of patients with NODAT without increasing the risk of acute graft rejection.

Blood Glucose ; Calcineurin Inhibitors ; therapeutic use ; Diabetes Mellitus ; Graft Rejection ; prevention & control ; Humans ; Hypoglycemic Agents ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; Sirolimus ; therapeutic use

OBJECTIVETo identify the proteins involved in osteogenic differentiation of beagle dog bone marrow mesenchymal stem cells (BMSCs) and explore its possible regulation mechanism of osteogenic differentiation of BMSCs.

METHODSCultured beagle dog BMSCs were induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) for 7 days. The differentially expressed proteins between cells with osteogenic differentiation and control cells were identified by proteomics analysis based on two-dimensional gel electrophoresis. Q-PCR and Western blotting were used to verify the interested protein of LASP1, ferritin light chain and heavy chain.

RESULTSOsteogenic differentiation was induced successfully in the BMSCs. Twenty differentially expressed proteins were identified by proteomic analysis, including 9 down-regulated and 11 up-regulated ones. Q-PCR and Western blotting demonstrated a significant reduction of LASP1 expression and significant up-regulation of ferritin in the BMSCs after a 7-day induction with rhBMP-2.

CONCLUSIONLASP1, which plays an important role in the regulation of the activity of the cytoskeleton, and ferritin, an important molecule in cellular iron homeostasis, can be critical in the osteogenic differentiation of beagle dog BMSCs induced by rhBMP-2.

Adaptor Proteins, Signal Transducing ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Bone Morphogenetic Protein 2 ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Cytoskeletal Proteins ; metabolism ; Dogs ; Down-Regulation ; Ferritins ; metabolism ; LIM Domain Proteins ; metabolism ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Osteogenesis ; drug effects ; Proteomics ; Recombinant Proteins ; pharmacology ; Transforming Growth Factor beta ; pharmacology ; Up-Regulation

Objective To develop Chinese version of the Manchester patient safety framework, and evaluate the feasibility of its clinical application. Methods The Manchester patient safety framework (MaPSaF) was transformed into Chinese version by translation and back-translation together with inquiries from experts. The trial study was done firstly among 136 nurses in five pilot wards and followed by five times of focus-oriented group interviews. A semi-structured, in-depth interview was then performed in 12 randomly selected participants and acquired data were analyzed by category analysis. Result Two themes were abstracted: Chinese version of MaPSaF showed a good practicability, effectiveness and operability and it was effective for deepened understanding of patient′s safety culture. Conclusion The Chinese version of MaPSaF can be used for safety culture evaluation during nursing process in China.