With the growth of people's living standard,the prevalence of childhood obesity is increasing year by year.Meanwhile,the adverse effect caused by obesity occurred more frequently as well.Therefore,obesity in childhood has become a critical issue in public health area.Complications of obesity may lead to enormous implications on children,including their endocrine,cardiovascular,digestive system,etc.,which will have an impact on the body growth in the adolescence.Obesity not only has a huge threat to children's physical and psychological health,but also has a long-term effect on body health condition in the future.Consequently,it is significant to diagnose the occurrences of complications of children,and implement the guiding and curing at an early stage.

Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P

A fused functional gene of human OPG and Mhsp65 was amplified by PCR,and cloned into the prokaryotic expression vector pET-28a.The BL21(DE3) strain of E.coli was transformed using the recombinant plasmid pET-28a-OPG-HSP65 and the expected protein was expressed by induction with IPTG.Result of SDS-PAGE indicated that the expected recombinant protein of 23 kDa was expressed with high yield as inclusion body.The fusion protein could be specifically recognized by both the anti-His antibody and anti-human OPG monoclonal antibody in Western blot analysis.The purified and refolded fusion protein could inhibit osteoclast proliferation and bone absorption in vitro.The results of mouse ear swelling assay and expressions of TNF-?,IFN-? and IL-17 mRNAs detected by real-time quantitative PCR demonstrated that the fusion protein had an anti-inflammation activity.The results suggest that the fusion protein of human OPG and Mhsp65 may act as a potential therapeutics for rheumatoid arthritis.

Huperzine A is a potent,reversible,and blood-brain barrier permeable acetylcholinesterase irhibitor.The aim of this study was to compare the pharmacokinetics,tolerability,and bioavailability of two formulations with the established reference formulation of huperzine A in a fasting,healthy Chinese male population.This was a randomized,single-dose,3-period,6-sequence crossover study.The plasma concentrations of huperzine A were determined by liquid chromatography tandem mass spectrometry.Tolerability was assessed based on subject interview,vital sign monitoring,physical examination,and routine blood and urine tests.The mean (SD) pharmacokinetic parameters of the reference drug were Cmax,1.550 (0.528) ng/mL;t1/2,12.092 (1.898) h;AUC0-72h,17.550 (3.794) ng.h/mL.Those of the test formulation A and test formulation B were Cmax,1.412 (0.467),1.521 (0.608) ng/mL;t1/2,12.073 (2.068),12.271 (1.678) h;AUC0-72h,15.286 (3.434) ng.h/mL,15.673 (3.586) ng.h/mL.The 90％ confidence intervals for the AUC0-72h and Cmax were between 0.80 and 1.25.No adverse events were reported by the subjects or found with results of clinical laboratory test.The test and reference products met the regulatory criteria for bioequivalence in these fasting,healthy Chinese male volunteers.All three formulations appeared to be well tolerated.

AIM To investigate the effects of asiatic acid (AA) on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD)-like motor symptoms in mice and its neuroprotective mechanism.METHODS Forty-five male C57BL/6 mice,except the nine mice in control group,were induced to be the PD models by peritoneal injection of 25 mg/kg MPTP for seven days and then were randomly assigned to model group,low-dose,high-dose AA groups and positive control group.Both the control group and the model group were administered with 0.5％ sodium carboxyl methyl cellulose (CMC-Na) solution,the AA groups were dosed with 12.5 mg/kg and 25 mg/kg AA,respectively,and the positive control group was given 75 mg/kg daily intragastric gavage of levodopa for eleven days.On the twelfth day,behavioral tests were performed.Tyrosine hydroxylase (TH) positive cells in substantia nigra were detected by immunohistochemistry.The mRNA expressions of iNOS,COX-2,TNF-α,IL-1β,and malonaldehyde (MDA) content in midbrain were measured.The levels of IL-1 β and TNF-α in the serum were detected using ELISA kits.RESULTS The mice treated with asiatic acid performed better in behavior tests than those in the model group (P ＜0.05,P ＜0.01).In addition,asiatic acid effectively protected the dopaminergic neurons in the substantia nigra due to upregulated TH expression and increased number of TH positive cells (P ＜ 0.05).The asiatic acid-treated mice had their mRNA expressions of IL-1β,TNF-α,iNOS and COX-2 in midbrain markedly suppressed (P ＜0.05,P ＜0.01),and a significant MDA level decrease in the midbrain tissue as well (P ＜ 0.01).Furthermore,reductions of IL-1 β and TNF-α contents in the serum were observed (P ＜ 0.05,P ＜ 0.01).CONCLUSION Asiatic acid attenuates motor dysfunction and dopaminergic neuronal deficits in PD mice,and the neuroprotective mechanisms may attribute to its anti-oxidative and anti-inflammatory activities.

OBJECTIVE: To explore the influence of visual attention in visual evoked potential (VEP) examination. METHODS: The pattern visual evoked potential (PVEP) and flash visual evoked potential (FVEP) were used to examine 110 normal subjects whose visual acuity or corrected visual acuity is in the range of 4.8-5.2, and age between 20 to 30 years old. In PVEP inspection, subjects were asked to watch with three ways, to stare at the central screen (at the same time count silently the number of alternating of black and white squares to improve the subjects' attention), to stare at the central screen at the same time thinking other things, to look sideways (the left eye stared at the left bottom corner of the screen or the right eye stared at the right one). In FVEP inspection, subjects were checked with eyes open and closed respectively. The latency and amplitude of P100 wave were recorded. The data were statistically analyzed by SPSS 13.0 software. RESULTS In PVEP inspection, the latency and amplitude of P100 wave were statistically different among three watching ways. In FVEP inspection, latency and amplitude of P100 wave were statistically different between eyes open and closed group. CONCLUSION Although the visual evoked potential test is an objective electrophysiological visual function test, it is susceptible to the influence of various subjective factors such as visual attention. It should arise the attention of forensic experts.
Adult ; Attention ; Cooperative Behavior ; Electroencephalography ; Electrophysiology ; Evoked Potentials, Visual/physiology* ; Female ; Forensic Medicine/methods* ; Humans ; Male ; Pattern Recognition, Visual ; Photic Stimulation ; Reaction Time ; Visual Acuity/physiology* ; Visual Fields/physiology* ; Young Adult

OBJECTIVETo study the effect of DPC4 gene expression on the growth of colon cancer cells and its mechanism.

METHODSExpression plasmid pcDNA3.1-DPC4 was constructed and transfected into the colon cancer cell line SW620 by use of lipofectamine gene transfer technique. DPC4 protein expression was detected by Western blot and immunocytochemistry. The effect of DPC4 gene on the growth of SW620 cells was monitored by population doubling time (PDT) and cloning efficiency. The influence of DPC4 expression on p21WAF1 transcription was investigated by RT-PCR to detect p21WAF1 mRNA.

RESULTSSuccessful expression of DPC4 protein was detected in the transfected SW620 cells. Compared with the control cells, PDT (74 h) of the DPC4 expressing cells was prolonged and the cloning efficiency (21%) decreased. In addition, the mRNA level of p21(WAF1) in DPC4 transfected cells was increased.

CONCLUSIONSOverexpression of DPC4 gene inhibits the growth of colon cancer in vitro, and induction of p21(WAF1) expression may be an important functional aspect of DPC4.

Carcinoma ; genetics ; metabolism ; pathology ; Cell Cycle Proteins ; biosynthesis ; genetics ; Cell Division ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p21 ; DNA-Binding Proteins ; biosynthesis ; genetics ; Gene Transfer Techniques ; Genes, Tumor Suppressor ; Humans ; Smad4 Protein ; Trans-Activators ; biosynthesis ; genetics ; Transfection ; Tumor Cells, Cultured

Animals ; Cardiomyopathies ; therapy ; Disease Models, Animal ; Doxorubicin ; toxicity ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Rabbits ; Transplantation, Autologous

OBJECTIVE: To explore the effect of Twist gene on the migration and invasion of human gastric carcinoma cells. METHODS: MKN28 cells, a human gastric carcinoma cell line, were transfected with PcDNA3.1-Twist plasmid by lipofectamine transfecting technique. The transfected cells were selected with geneticin. Expressions of Twist,ecadherin and vimentin protein were detected by Western blot in cells transfected Twist gene. Matrigel invision chambers were performed to analyse the cell migration and invasion. RESULTS: MKN28 cells transfected with PcDNA3.1-Twist plasmid showed stronger intracellular expression of Twist protein than MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The expression of ecadherin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly decreased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without the transfection. However, The expression of vimentin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly increased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The migration and invasion ability of Twist+ - MKN28 cells were stronger than that of MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. CONCLUSION Twist gene may promote the migration and invasion ability of gastric carcinoma cells through epithelial mesenchymal transition.
Cadherins ; biosynthesis ; Cell Movement ; genetics ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection ; Tumor Cells, Cultured ; Twist-Related Protein 1 ; biosynthesis ; genetics ; Vimentin ; biosynthesis

OBJECTIVETo investigate the bacteriological character and pH value in nasal cavity and observe the efficacy following 8 week treatment with Budesonide aqueous nasal spray in chronic nonallergic rhinitis.

METHODSForty-two patients with chronic nonallergic rhinitis were treated with Budesonide aqueous nasal spray (256 microg once daily intranasal) for 8 weeks. Bacteriological character was measured before and 8 weeks after treatment All the swab specimens were obtained from the inferior meatus, bacteriologic outcome was determined by general cultures. Nasal pH value was measured by using a probe sited under the inferior turbinate before and after treatment. Efficacy was evaluated by measurement of nasal symptom scores and sign scores before and 8 weeks after treatment.

RESULTSThe bacterial growth was present in 37 of 42 cases (88%) before the treatment (group 1), including 53 aerobic bacteria. The bacterial growth was present in 38 of 42 cases (90.47%) 8 weeks after treatment (group 2), including 46 aerobic bacteria. There was no significant difference in the bacterial distribution between group 1 and group 2 (chi2 = 0.416, P > 0.05). Budesonide aqueous nasal spray reduced pH value from 7.90 +/- 0.39 to 7.70 +/- 0.23 (t = 2.72, P < 0.05). All the parameters of symptoms were improved after treatment, including nasal obstruction, nasal secretions, itchy feeling of the nose, closed rhinolalia and headache.

CONCLUSIONSGlucocorticoid aqueous nasal spray could be used safely for eight weeks and does not increase the risk of aerobes and fungi infection and reduce the pH in chronic nonallergic rhinitis. It is effective in relieving symptoms of patients with chronic nonallergic rhinitis.

Administration, Intranasal ; Adult ; Aged ; Budesonide ; administration & dosage ; therapeutic use ; Chronic Disease ; Female ; Humans ; Hydrogen-Ion Concentration ; Male ; Middle Aged ; Nasal Cavity ; microbiology ; Rhinitis ; drug therapy ; microbiology ; physiopathology