Aim To investigate the effect of paeoniflorin(PF)on TLR2/4 pathway in AGEs-induced RAW264.7 macrophages.Methods RAW264.7 macrophages were incubated at different time points in AGEs stimulation,as well as different concentrations of PF,to optimize experimental conditions.RAW264.7 macrophages were randomly divided into five groups: control group(DMEM),bull serum albumin(BSA)group(200 mg·L-1 BSA),AGEs group(200 mg·L-1 AGEs),paeoniflorin group(200 mg·L-1 AGEs+10-5 mol·L-1 PF)and TLR2/4 inhibitor group(200 mg·L-1 AGEs+30 mg·L-1 OxPAPC).The expression of Toll-like receptor 2(TLR2),Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),p-IRAK1,TIR-domain containing adaptor protein-inducing IFN-β(TRIF),interferon regulatory factor 3(IRF3),p-IRF3,NF-κB p-p65,NF-κB p65,inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α),interleukin-l β(IL-1β)and monocyte chemotactic protein-1(MCP-1)were measured by Western blot.Real-time PCR was used to detect the expression of TLR2 and TLR4 mRNA,while TNF-α,IL-1β and MCP-1 levels in cell supernatant were measured by ELISA.Results Compared with control group,AGEs significantly increased the expression of TLR2,TLR4,MyD88,p-IRAK1,TRIF,IRF3,p-IRF3,NF-κB p-p65,NF-κB p65,iNOS,TNF-α,IL-1β and MCP-1 proteins(P<0.01),as well as TLR2 and TLR4 mRNA(P<0.01).TNF-α,IL-1β and MCP-1 contents were also elevated in cell supernatant(P<0.01).The effects induced by AGEs were decreased significantly in PF and TLR2/4 inhibitor group(P<0.01).Conclusion PF plays an anti-inflammatory effect via inhibiting TLR2/4 pathway on macrophages,which may provide a new theoretical basis for the treatment of diabetic nephropathy.

Objective To evaluate the clinical effects of ultrasound-guided internal jugular vein catheterization in long axis plane,short axis plane and oblique axis plane,in order to identify the opti-mal axis plane for this procedure.Methods One hundred and eighty patients (male 94 cases,female 86 cases,aged 34-82 years)requiring ultrasound-guided internal jugular vein catheterization were in-cluded in this study.They were randomly divided into three groups (n =60 each),long axis group, short axis group and oblique axis group,with 60 cases in each group.The details of catheterization in-cluding the time accessing into vein,the time finishing cannulation,needle redirecting times,number of skin points of puncture,puncture successful rate and complications in the three groups were recor-ded.Results Compared with long axis plane and short axis plane,the oblique axis plane was associat-ed with decreased time for venous access and cannulation.The oblique axis plane also needed less changes of needle direction.The complication of arterial puncture in the oblique axis plane group was significantly lower than long axis plane group and short axis plane group(P <0.05).The number of skin puncture points were similar between the three groups.Conclusion The oblique plane can provide a safe and more effective route to perform the IJV catheterization with minimal risk for carotid artery puncture,which demonstrates the practical superiority over the classic short axis plane and long axis plane for critically ill patients.

OBJECTIVETo observe the effects of H. dulcis on relieving alcohol toxicity by animal experiments.

METHODMale kunming mice were ovraiectomized and randomly divided into 5 groups: control group, model group, and aqueous extracts of H. dulcis group at 0.5, 0.25, 0.125 g x mL(-1). The acute alcohlism animals induced by gastral administration with "Er Guo Tou" and the alchol concentrations in serum were detected after treated with the extracts within 0.5-3 h by biochemical enzymes.

RESULTThe alcohol concentration in blood was up to the maximum in 0.5-1.5 h. However, the alcohol concentrations in blood of aqueous extract from H. dulcis group were decreased in 0.5-3 h. The activity of ADH in the liver in aqueous extract of H. dulcis group was increased in 2-3 h, while it was significantly increased in 1-1.5 h (P <0.05).

CONCLUSIONThe aqueous extract of H. dulcis could reduce the alchol concentration in blood of animals and inrease the activity of ADH after given alcohol. It means the extract has the effect of relieving alcohol toxicity and preventing drunkenness through restraining the absorption of alcohol in the gastrointestinal tract and promoting the metabolism of alcohol in the liver.

Alcohol Dehydrogenase ; metabolism ; Animals ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ethanol ; blood ; toxicity ; Liver ; enzymology ; Male ; Mice ; Mice, Inbred ICR ; Plants, Medicinal ; chemistry ; Random Allocation ; Rhamnaceae ; chemistry ; Seeds ; chemistry

Objective To determine the influence of P-glycoprotein (P-gp) inhibitor on the blood brain barrier (BBB) transport of amphotericin B (AmB)..Methods An in-vitro BBB model was established with brain capillary endothelia cells (BCEC). AmB was chosen as the test drug and verapamil was chosen as the inhibitor of P-gp.Cellular uptake of AmB at different time points and with series of verapamil concentrations were performed respectively after the determination of appropriate incubation time and drug dosage by the cytotoxicity assay. The AmB concentrations of series of samples were detected using high performance liquid chromatography (HPLC) method. One-way ANOVA analysis and Bonferroni test were used for data analysis.Results The cellular transport of AmB was accumulated as the time prolonged.The inhibitor group had a significant higher cellular uptake levelsof AmBat the time point of 90 min (t=6.753,P=0.001),120 min (t=3.574,P=0.016) and 150 min (t=4.759,P=0.005) as compared with the control group.The AmB cellular uptake level increased significantly when BCEC were incubated with verapamil of 2 μmol/L (P=0.000),5 μmol/L (P=0.014),10 μmol/L (P=0.000),50 μmol/L (P=0.014),75 μmol/L (P=0.000) and 100 tμmol/L (P=0.000),respectively,compared with the control group.Conclusion The P-gp inhibitor verapamil can enhance the cellular uptake of AmB which indicates that P-gp is involved in the BBP transport of AmB.

Objective: To obtain the bFGF mimic peptide binding to FGFR via phage display, and to provide the base for developing peptide agonist of bFGF. Methods: Using Balb/c 3T3 cells as the target cells and COS-7 cells as the subtractive panning, the phage display heptapeptide library was biopanned for 4 rounds to obtain the single phage clones. The affinity and the specificity of the clones were assessed by ELISA. DNA sequencing was applied to further analyze the positive clones. Results: Twelve positive clones were selected from the enriched phages. A group of hydrophobic peptides containing a conserved motif, PR, was identified. Conclusion: Two bFGF mimic heptapeptides binding to FGFR were selected, which may be used as the candidates for bFGF agonist.

OBJECTIVETo summarize the clinical manifestation and diagnosis of ganglioneuroma in spine and investigate the clinical effect of surgical treatment.

METHODSThe clinical data of 6 patients underwent a surgery for ganglioneuroma in spine from January 2008 to January 2015 were retrospectively analyzed. There were 4 males and 2 females, aged from 2 to 63 years old with an average of 34.6 years. The courses of disease were from 3 days to 17 years. Five patients complicated with superficial hypesthesia in correlative level of tumor, and the muscle strength under tumor plane had decreased at different levels, with the strength of grade II-IV. Two cases complicated with hypermyotonia and positive bilateral Hoffmann's and Babinski sign. Five cases were sporadic lesion in correlative spinal canal and one case complicated with the giant occupying lesion in thoracic cavity.

RESULTSSix operations had been performed including 5 en bloc and 1 subtotal resection. Postoperative pathological results showed tumor cells scattered or fasciculated inserted into Schwann cells in the stroma. In 2 patients complicated with radiculalgia before operation, 1 case was relieved and 1 was invariant after operation. All 4 patients with preoperative dyscinesia in the limbs obtained improvement after operation. All the patients were followed up from 0.3 to 6.8 years with an average of 2.5 years. At the final follow-up, according to ASIA grade, 5 cases were good and 1 case was invariant. During the follow-up, only 1 patient experienced chemoradiation because of merging ganglioneuroblastoma and receiving subtotal resection. No recurrence in other 5 cases.

CONCLUSIONGanglioneuroma is a benign and rare tumors in spine. Clinically, radicular pain and sensory-motor disorders are the main manifestations. Its diagnosis depends on pathological examination. Prognosis of surgical treatment is good.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Ganglioneuroma ; diagnosis ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spinal Neoplasms ; diagnosis ; surgery

To investigate the mechanism of inhibitory effect of a novel bFGF antagonist peptide isolated from the phage display random heptapeptide library on cell proliferation induced by basic fibroblast growth factor. The effect of P7 on cell morphology was observed under an inverted microscope. Flow cytometry was applied to analyze the effect of P7 on cell cycle progress of bFGF-stimulated cells. The effect of P7 on bFGF-induced activation of MEK and Erk1/2 in MAPK pathway was detected by Western blotting. The results showed that no significant cell morphology change was observed in the range of detected concentrations of P7. Cell cycle analysis showed that P7 decreased S-phase cell population and arrested cell cycle at the G0/G1 phase of bFGF-stimulated cells. The results of MAP kinase activation assay indicated that P7 decreased bFGF-induced MEK and Erk1/2 phosphorylation in a dose-dependent manner. P7 inhibited proliferation of bFGF-stimulated Balb/c 3T3 cells possibly via cell cycle arrest at the G0/G1 phase and down-regulation of signal molecular activation in MAPK pathway.
Animals ; BALB 3T3 Cells ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Fibroblast Growth Factor 2 ; antagonists & inhibitors ; pharmacology ; MAP Kinase Kinase Kinases ; metabolism ; MAP Kinase Signaling System ; drug effects ; Mice ; Mice, Inbred BALB C ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Peptides ; pharmacology ; Phosphorylation ; Protein Binding

OBJECTIVETo label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects.

METHODSGFP cDNA was inserted into pcDNA(3.1)-hIGF-1 to construct the co-expression vector with two multiple cloning sites mammalian expression vector under two cytomegalovirus promoters/enhancers respectively. Recombinant pcGI was transfected into NIH 3T3 cells with the help of lipofectamine.

RESULTSEnzyme digestion and agarose gel electrophoresis analysis revealed that pcGI vector contained correct GFP and hIGF-I cDNA. Expression of hIGF-1 and GFP was confirmed in transfected NIH 3T3 cells by immunocytochemical analysis and fluorescence microscopy.

CONCLUSIONShIGF-I eukaryotic expression vector has been successfully labeled with GFP.

Cartilage, Articular ; injuries ; Cells, Cultured ; Eukaryotic Cells ; cytology ; physiology ; Fibroblasts ; cytology ; physiology ; Gene Expression Regulation ; Genetic Therapy ; methods ; Genetic Vectors ; Green Fluorescent Proteins ; pharmacology ; Humans ; Immunohistochemistry ; Insulin-Like Growth Factor I ; genetics ; therapeutic use ; Luminescent Agents ; pharmacology ; Sensitivity and Specificity ; Transfection ; methods ; Wounds and Injuries ; diagnosis ; therapy

OBJECTIVETo explore the curative effects and complications of elastic intramedullary nail in treating children's bilateral femoral shaft fractures.

METHODSForm February 2005 to March 2008, 7 patients with bilateral femoral shaft fractures were treated by closed reduction and internal fixation with elastic intramedullary nail. There were 5 males and 2 females. The age ranged from 3 to 13 years with the mean of 8.3 years. Six injuries caused by road accident and 1 injury caused by fall from high. Two cases associated with pulmonary contusion, 3 cases brain injuries, 1 case fracture of calcaneus and 1 case bladder injuries. All the cases were closed fractures without nerve and blood vessel injury. A cast external fixation had been used after operation for a month in two cases.

RESULTSAll the patients were followed up for 21-37 months with an average of 30.3 months. No infecton of incisional wound, displacement fracture, internal fixation fail, delayedunion and malunion were found. All fracture obtained healing for 7-12 weeks with an average of 8.7 weeks. Inequality of lower limb was found in 1 case (length differences was 5 mm). According to Flynn scoring,all fractures were excellent.

CONCLUSIONTreatment of femoral shaft fractures in children with elastic intramedullary nail according with biological principle. The method has little trauma, less complication, outstanding effect and it is a good way to treat bilateral femoral shaft fractures result from high-energy injuries.

Adolescent ; Bone Nails ; Child ; Child, Preschool ; Elasticity ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Intramedullary ; adverse effects ; methods ; Humans ; Male

BACKGROUNDConstruction of replication-deficient recombinant adenovirus expressing gag-pol and env genes of human immunodeficiency virus (HIV) in mice.

METHODSgag-polDelta and gp140TM genes were cloned into shuttle vector pAdTrack-CMV respectively, and then the plasmids containing gag-polDelta or gp140TM gene were cotransformed with the backbone of adenovirus into E.coli BJ5183. Transfections of the recombinants were performed to obtain recombinant adenoviruses. Its immunogenicity was evaluated by testing antibody levels of mice primed with DNA vaccines and boosted with recombinant adenoviruses.

RESULTSThe replication-deficient recombinant adenovirus could express Gp140TM, Gag P55 and P24 proteins correctly. The mice primed with DNA vaccines and boosted with recombinant adenoviruses elicited high titer of HIV-1-specific antibody compared with that inoculated with DNA vaccines only.

CONCLUSIONReplication-deficient recombinant adenovirus expressing gag-polDelta and gp140TM can elicit high titer HIV-1-specific antibodies.

AIDS Vaccines ; immunology ; Adenoviridae ; genetics ; Animals ; Female ; Fusion Proteins, gag-pol ; biosynthesis ; genetics ; Gene Products, env ; biosynthesis ; genetics ; HIV-1 ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Recombination, Genetic ; Transfection ; Vaccines, DNA ; immunology ; env Gene Products, Human Immunodeficiency Virus