Animals ; Aorta, Thoracic ; physiology ; Endothelium ; blood supply ; Male ; Physical Conditioning, Animal ; physiology ; Rats ; Rats, Sprague-Dawley ; Vasodilation

Aged ; Diagnostic Errors ; Humans ; Lung Neoplasms ; diagnosis ; Male ; Middle Aged ; Pneumonectomy ; Silicosis ; diagnosis ; surgery

Objective To investigate the changes of CD4~+ T cell subset in the role of immuno-pathogenesis of type 1 diabetes mellitus(T1DM). Methods Real-time PCR was used to evaluate the ex-pression levels of transcriptional factors (T-bet, GATA-3, Foxp3, ROR-γt), cytokines (IFN-γ, IL-4, IL-10, IL-17A) and negative regulatory factors (CTLA-4, GITR) mRNA from CD4~+ T cells. The proportions of Th1, Th2, Tr and Th17 cells in peripheral blood were detected by flow cytometric analysis. Plasma cyto-kine (IFN-γ, IL-4, TGF-β and IL-6) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Results (1) The proportions of Th1 cells in peripheral blood from children with T1DM were siguificanfly increased than that of healthy controls, and proportions of Th2 were decreased (P < 0.01). There were no significant differences between diabetic patients and healthy controls regarding the proportions of Tr cells and Th17 cells(P >0.05). (2) Transcription levels of T-bet and IFN-γ mRNA were significantly up-regulated, while GATA3 and IL-4 were significantly down-regulated in children with T1DM. The mRNA expression levels of Tr negativity regulatory factors such as IL-10, CTLA-4 and GITR were lower in CD4~+ T cells from children with TIDM compared with the controls(P <0.01). There were no statistically differences to be observed in mRNA expression levels of ROR-γt and IL-17A genes between two groups(P > 0.05).(3) In comparison with controls, serum concentrations of IFN-γ or IL-4 were remarkable increased or de-creased respectively (P < 0. 01), while TGF-β and IL-6 did not change in children with T1DM (P > 0.05). Conclusion The Th1/Th2 imbalance might be play an important role in immunopathogenesis of T1DM. Functional deficiency of Tr cell might further exacerbate Th1/Th2 imbalance and lead to disturbance of cellu-lar immune response.

Objective: To determine protein binding characteri stic and signal transmission pathway of melatonin（Mel） receptor(MR) in human e mbryonic peripheral organ tissues. Methods: MR was measured by radio ligand-binding assay and the effect of GTPγS on melatonin specific bindi ng was studied. Results: Mel specific binding sites were det ermined in 16 kinds of human embryonic tissue and this binding could be inhibit ed by GTPγS, supporting the theory that MR is coupled to inhibitory G-proteins system. Conclusion: MR is measured in human embryo tissue, the se results provide experimental data for elucidating the mechanism of the effect of Mel.

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To evaluate the expression of Toll-like receptor(TLR)on the monocyte- derived dendritic cells(DC)from chronic hepatitis B(CHB)patients and to analyze the expression pro- file and significance of the TLR such as TLR3,TLR4,TLR?,TLR8 and TLRg,which are associat- ed with immune response to viral infection.Methods Peripheral blood mononuclear cell(PBMC) centrifugated by the hydroxyethyl starch(HES)centrifugation were cultured and induced into DC by granulocyte-maerophage colony stimulating factor(GM-CSF)and interleukin-4(IL-4),and their mor- phology and phenotype were detected by the inverted microscope and flow cytometry respectively. Monocyte-derived DC were obtained from 10 chronically hepatitis B virus(HBV)-infected patients and 15 healthy volunteers.TLR3,TLR4,TLR7,TLRS,TLR9 expression on immature and mature DC were analyzed by FACS Calibur.DC was pulsed with HBcAg on day 3 and 5,then DC maturation and ability to process HBcAg and to stimulate autogeneic T cells were evaluated.Results Monocyte- derived DC developed different TLR expression patterns as they went through different maturation stages.TLR7,TLR8 expressions on immature DC and TLR3,TLR7 expressions on mature DC were lower in CHB than in control(for TLR7,TLR8 expression on immature DC:75.9%,1.0%vs 98.4%,15.4%,P

OBJECTIVETo explore the technique and therapeutic effect of bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral shaft fractures in young children. Methods:From June 2006 to June 2012,45 young children with femoral shaft fractures were treated by bridge wire splint fixation with ankle dorsiflexion,which was designed according to arch bridge mechanical principle and structure. There were 31 males and 14 females with an average age of 3.2 years old ranging from 8 months to 5.5 years old; 14 cases were upper 1/3 femoral fractures,26 cases were middle 1/3 femoral fractures,5 cases were lower 1/3 femoral fractures; 20 cases were transverse fractures, 14 cases were oblique fractures,6 cases were spiral frac- tures, and 5 cases were comminuted fractures. X-ray, follow-up imaging changes,clinical curative effect and complications were assessed.

RESULTSForty-five patients were followed up for 6 to 21 months (averaged 12 months). All fractures were reached clinical bone healing after 5 to 7 weeks (averaged 6 weeks) fixation. Seven cases appearred limb soft tissue complications, including buttocks bedsore,dorsal foot and Achilles tendon epidermal necrosis, and healed after dressing and removal of external fixation. During follow-up,the original overlap angle and lateral displacement were remodeled, and limbs were restored to the normal line of force and bone structure. According to Flynn standard, 35 cases got excellent results, 8 cases good, 2 cases fair.

CONCLUSIONThe bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral. shaft fractures in young children (less than 6 years old) is safe,feasible, simple,and has raliable effect, which can be applied in primary hospitals.

Ankle ; surgery ; Bone Wires ; Child ; Child, Preschool ; Female ; Femoral Fractures ; surgery ; Femur ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; instrumentation ; Humans ; Infant ; Male ; Treatment Outcome

OBJECTIVETo study the antiproliferation effect on HepG2 cells and its underlying mechanism of the active chemical composition of the Viburnum Odoratissimum.

METHODS3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and trypan blue dye exclusion assay were used to assess the effect of vibsane-type diterpenoids on the proliferation of various tumor cells. Alterations in cell cycle and apoptosis were determined by flowcytometry. The enzymatic activity of caspase-3/7 was measured by Apo-ONE homogeneous Caspase-3/7 Assay kit.

RESULTSCompound 1 #, a vibsane-type diterpenoid, was found to significantly inhibit the growth of HepG2 cells by anticancer proliferation activity screening. It was demonstrated that the modified groups on side chain coupled to C11 site affected the cell growth-inhibition activity of compounds by structure-activity analysis. In addition, HepG2 cell line was most sensitive to compound 1 #, which induced growth arrest of HepG2 cells in a dose- and time-dependent manner. Study on the mechanisms underlying these effects indicated that compound 1 # induced significant G0/G1 phase arrest of HepG2 cells in a time- and concentration-dependent manner. Meanwhile, It was found that higher concentrations of compound (5-10 micromol/L) caused evident increase in the unmber of apoptotic cells and dose-dependent activation of caspase-3/7.

CONCLUSIONVibsane-type diterpenoids could significantly inhibit the growth of HCC HepG2 cells. Induction of cell cycle arrest and apoptosis may play important roles in their anticancer effects.

Apoptosis ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; Hep G2 Cells ; Humans ; Viburnum ; chemistry

OBJECTIVETo investigate the stress shielding after mini-plate internal fixation for mandibular fractures.

METHODSEighteen patients with mandibular fractures were selected.X-rays were taken before operation and 3, 4, 6 months after operation when the plates were removed. The bone density of the mini plate area was examined and compared among different time interval groups.

RESULTSThe bone density before operation (125.41 ± 2.47) and 3 months after operation (120.19 ± 3.52) was not significantly different, but became lower 4 and 6 months after operation than before operation.

CONCLUSIONSThere appeared stress shielding after internal fixation for 4 and 6 months in mandibular fracture, and the mini plate should be removed at those times.

Bone Density ; Bone Plates ; Fracture Fixation, Internal ; Humans ; Mandibular Fractures ; surgery

Objective: To determine protein binding characteri stic and signal transmission pathway of melatonin（Mel） receptor(MR) in human e mbryonic peripheral organ tissues. Methods: MR was measured by radio ligand-binding assay and the effect of GTPγS on melatonin specific bindi ng was studied. Results: Mel specific binding sites were det ermined in 16 kinds of human embryonic tissue and this binding could be inhibit ed by GTPγS, supporting the theory that MR is coupled to inhibitory G-proteins system. Conclusion: MR is measured in human embryo tissue, the se results provide experimental data for elucidating the mechanism of the effect of Mel.