Objective To investigate the gene expression of connexin43(Cx43) and its effect on gap junction intercellular communication(GJIC) of acute leukemia bone marrow stromal cells(ALBMSCs).Methods After ALBMSCs were transfected by recombinant adenovirus Ad-Cx43-GFP,the expression of report gene GFP and the transfection efficiency were monitored by fluorescent microscopy.RT-PCR,Western blot and immunocytochemical method were used to detect the mRNA and protein expressions of Cx43.Dye transfer procedure was performed to examine the GJIC function.Results After transfected by Ad-Cx43-GFP for 24 h,the expression of GFP in ALBMSCs was detected by fluorescent microscopy and the transfection efficiency was(82.7?2.16)%;The mRNA and protein expressions of Cx43 in ALBMSCs transfected by Ad-Cx43-GFP were higher than those not transfected by Ad-Cx43-GFP(P

Epidermodysplasia verruciformis (EV), a rare inherited disease, is believed to be associated with human papillomavirus (HPV) infection. EVER1/2 genes, dendritic cells, T lymphocytes, and the biological characteristics of HPV itself may play roles in the pathogenesis of HPV infection.
Dendritic Cells ; immunology ; Epidermodysplasia Verruciformis ; genetics ; immunology ; virology ; Humans ; Membrane Proteins ; genetics ; Mutation ; Papillomaviridae ; isolation & purification ; pathogenicity ; Papillomavirus Infections ; complications ; T-Lymphocytes ; immunology

Objective To improve objectivity and accuracy of the diagnosis,prognosis,treatment efficiency and observe the levels of cognitive and intelligent deficits of children with attention deficit hyperactive disorder(ADHD).Methods Event related potentials(ERP)P3 wave test provocated by audition and Wechsler intelligence scale for children(C-WISC) test were detected and compared in 60 children with ADHD(ADHD group) and 60 cases of healthy children(healthy control group).The ERP P3 wave test results between 2 groups of children which had different intelligent balance ability were also compared.Results Compared with the healthy control group,there was a significantly longer latency of P3 wave(P

BACKGROUND: The surface electromyography can evaluate the effect of exercise by recording the parameters of muscle activities, and vibration exercise is the best supplement to traditional weight training. More research focuses on the effect of simple vibration training on the surface electromyography of lower limbs, but the influence of vibration combined with weight-bearing training is poorly understood.OBJECTIVE: To study the effect of vibration combined with load on the surface electromyography of lower limbs at the micro level of muscle working principle.METHODS: Eight healthy college students were recruited, and subjected to four different stimulations: vibration (45 Hz) combined with load (45% one repetition maximum (IRM)); vibration (45 Hz) combined with load (60% 1RM); vibration (50 Hz) combined with load (45% 1RM); vibration (50 Hz) combined with load (60% 1RM), followed by semi-squat exercise with the heel lifting, 10 times/minute, for 3 courses with more than 2 hours in between.RESULTS AND CONCLUSION: Different vibrations combined with loads made significant difference on the root mean square amplitude of the surface electromyography (P < 0.01), and there was a significantly increased root mean square amplitude in the vibration (50 Hz) combined with load (45% 1RM), especially at the medial gastrocnemius. The four kinds of stimulations made significant different effects on the surface electromyography of rectus femoris, tibialis anterior, and medial gastrocnemius, except semitendinosus (P ≤ 0.05). Moreover, the effect showed significant difference among different stimulations except vibration (50 Hz) combined with load (45% 1RM) (P ≤ 0.05). Compared with the other three stimulations, vibration (50 Hz) combined with load (45% 1RM) exerted better effect on the muscular activation. To conclude, different vibrations combined with loads exert different effects on the motor unit of same neuromuscular activity, and a suitable stimulation may produce better effect. Besides, the same stimulus for the motor unit of different neuromuscular activities produces different effects, which may match to the muscle nature.

OBJECTIVEThe objective of this study is to investigate the influence of mechanical properties and sintering performance by adding 5% weight percentage aids to nano-compound zirconia toughened alumina (ZTA) ceramics.

METHODSMicrometer Al2O3 and nanometer ZrO2 (quality ratio 4:1) were used to get 55% volume percentage slurry. Magnesium oxide and titanium oxide were taken as aids which were 5% weight percentage of the Al2O3 and ZrO2 powder. Five groups (number 0, 1, 2, 3, 4 group) were divided according to different proportion of aids. After gel-casting, the porcelain pieces were sintered at 1150, 1200, 1300, 1400, 1450, 1500, 1600 degrees C for 2 hours. Static three-point flexure strength, line shrinkage, relative density were measured and scanning electron microscopy (SEM) was used to observe section.

RESULTSNumber 1 (MgO 1%, TiO2 4%) group had the highest bending strength. It was (401.78+/-19.50) MPa after sintering at 1600 degrees C for 2 hours and was higher than 0 group (380.64+/-44.50) MPa. Bending strength became lower than 0 group when MgO was more than 2% or more than that weight percentage of ZTA powder. When MgO content was higher than 2% or more than that weight percentage, there was no difference in relative density raising rate between each sintering assistants groups. When the sintering temperature was higher than 1200 degrees C, all groups showed obvious line-shrinkage and the groups which contained sintering assistants were all was higher than 0 group.

CONCLUSIONAdding MgO and TiO2 aids from 1% to 4% weight percentage of ZTA will promote fritting and increase ZTA nano-compound ceramics mechanical properties. Adding 2% MgO aids or more than that weight percent will has no obvious help to increase the relative density raising rate of ZTA nano-compound ceramics and will degrade the mechanical properties of ZTA nano-compound ceramics.

Aluminum Oxide ; Dental Porcelain ; Microscopy, Electron, Scanning ; Temperature ; Titanium ; Zirconium

OBJECTIVE We have recently reported that cysteinyl leukotriene (CysLT) signaling plays an important role in microglial interleukin (IL)-1β secretion and subsequent neurotoxicity. The present study aimed to examine microglial morphological changes and the upstream molecular underlying IL-1βproduction in CysLT receptor agonist leukotriene D4 (LTD4)-treated BV2 microglia in vitro. METHODS Twenty-four hours after murine microglial BV2 cells were stimulated with LTD4 (1-100 nmol·L- 1), the cell proliferation and morphology were observed. The expression level of cysteinyl aspartate-specific protease 1 (CASP1) protein was measured by Western blotin BV2 cells. In addition, BV2 cells were pretreated with or without CysLT1 receptor antagonist montelukast for 1 h and the effects of monte-lukaston LTD4-stimulated microglial activation and CASP1 expression were evaluated. RESULTS The number of BV2 cells had an increasing tendency after 24 h treatment with LTD4, but no significant differences were observed between the control and LTD4-treated cells (P>0.05). Under basal and resting conditions, BV2 microglial cells displayed a ramified morphology. However, LTD4 at 100 nmool · L- 1 drove microglial morphological changes from a ramified towards an amoeboid shape. The expression of CASP1 protein was significantly upregulated in 100 nmool·L-1 LTD4-treated BV2 microglia (P<0.01). Furthermore, pretreatment with CysLT1 receptor antagonist montelukast prevented cell morphological changes and suppressed the increased CASP1 expression in LTD4-treated BV2 cells (P<0.05). CONCLUSION CysLT receptor agonist LTD4 induces morphological changes and CASP1 expressionin BV2 microglia, which can be inhibited by CysLT1 antagonist. These results suggest the involvement of CysLT signaling in microglial morphological changes and CASP1 expression.

OBJECTIVETo assess the anti-tumor immune response to percutaneous cryoablation in patients with local prostate cancer.

METHODSWe treated 10 patients with local prostate cancer by percutaneous cryoablation, collected the blood samples before and 2 weeks after the treatment and isolated peripheral blood mononuclear cells (PBMCs). Protein lysates were made by biopsy from autologous prostate cancer or non-cancer tissues. The levels of serum TNF-alpha, IFN-gamma, IL4 and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA) and the Th1/Th2 ratio was calculated by the IFN-gamma/IL-4 ratio. The number of IFN-gamma + T cells under the stimulation of different protein lysates was counted by enzyme link immunol spot (ELISPOT). And the cytolytic activity of cytotoxic T lymphocytes (CTL) was detected by LDH assay.

RESULTSCompared with pre-treatment, the levels of TNF-alpha and IFN-gamma, the Th1/ Th2 ratio and the number of IFN-gamma + T cells induced by tumor protein lysates in PBMCs were increased significantly after cryosurgery (P < 0.01), while the levels of IL4 and IL-10 decreased slightly, and the non-tumor protein lysates induced no obvious changes in the number of IFN-gamma T cells. The cytolytic activity of cytotoxic T lymphocytes against human prostate cancer cells LNCaP was markedly increased, but not that against renal cancer cells GRC-1. One case of recurrence was found during the 3-6 months follow-up.

CONCLUSIONPercutaneous cryoablation for prostate cancer could induce a tumor-specific immune response.

Aged ; Cryosurgery ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Male ; Middle Aged ; Prostatic Neoplasms ; immunology ; therapy ; T-Lymphocytes, Cytotoxic ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Necrosis Factor-alpha ; blood

BACKGROUNDHuman beta-defensin-3 (HBD(3)) is an epithelial peptide that has been demonstrated to have a salt-insensitive broad spectrum of potent antimicrobial activity. Expressing antimicrobial peptides in Escherichia coli (E. coli) is very difficult for it can result in death of the bacterial host cells. Our aim was to establish a prokaryotic system expressing soluble HBD(3) protein and demonstrate the antimicrobial activity of the expressed protein. We then studied whether the host cells would activate the suicide pathways.

METHODSWe first cloned the complementary DNA coding for the mature chain of HBD(3), inserted it into the vector PGEX-KG then transformed E. coli BL21 (DE3) with the appropriate recombinant plasmid. After induction with 0.5 mmol/L isopropyl-1-thio-beta-D-galactopyranoside (IPTG) the transformed E. coli produced a recombinant glutathione S-transferase and HBD(3) (GST-HBD(3)) fusion protein. The fusion protein was treated with thrombin to produce pure HBD(3) protein then the antimicrobial activity of HBD(3) was evaluated in a liquid microdilution assay.

RESULTSThe fusion protein GST-HBD(3) was efficiently cleaved by thrombin and yielded HBD(3) that had anti-staphylococcus aureus activity with a minimal inhibitory concentration level of 12.5 microg/ml. The E. coli strain expressing the recombinant protein did not grow slower than the empty vector strain.

CONCLUSIONActive HBD(3) in E. coli by expressing the recombinant protein GST-HBD(3) could be produced, and suicide did not occur in the E. coli strain expressing the recombinant protein.

Amino Acid Sequence ; DNA, Complementary ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; growth & development ; Glutathione Transferase ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Fusion Proteins ; chemistry ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Staphylococcus aureus ; drug effects ; Thrombin ; metabolism ; beta-Defensins ; genetics ; metabolism ; pharmacology

This study was aimed to develop and establish an efficient method for high through-put automatically extracting genomic DNA from EDTA-anticoagulated whole blood samples, and to utilize this method in routine rSSO HLA genotyping by luminex flow array assay, the genomic DNA was extracted automatically from 400 microl blood samples by using TECAN DNA workstation and 96-well plate with 2 ml volume per well. The yield and purity of each DNA sample was tested by UV-spectrophotometer, the integrity of these DNA samples were run electrophoresis on the agarose gel. Each DNA sample was subjected to PCR amplification and hybridization using One lambda rSSO HLA-A, -B and -DRB1 commercial kit, the fluorescent intensity for positive bead and negative bead hybridized with HLA-A, -B and -DRB1 PCR products were calculated and analyzed. The results showed that the mean yield and purity (A260/A280) of genomic DNA extracted from 400 microl whole blood samples were 3.217+/-0.715 microg and 1.710+/-0.103 respectively. The molecular weight was more than 15 kb in size and the fluorescent intensity for positive bead hybridized with HLA-A, -B and -DRB1 PCR products of each sample was >600 RFU, however, the fluorescent intensity for negative bead for each sample was <50 RFU. It is concluded that the highly qualified genomic DNA can be extracted automatically from blood samples of marrow-donors by using TECAN DNA workstation, and the extracted DNA samples are suitable for high through-put HLA genotyping by luminex flow array assay and other downstream transplant immunological and molecular biological experiments.
Biological Specimen Banks ; Bone Marrow ; DNA ; isolation & purification ; DNA Primers ; Genotype ; HLA Antigens ; genetics ; High-Throughput Screening Assays ; methods ; Humans ; Living Donors ; Nucleic Acid Hybridization ; methods ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo investigate the expressions of CD1a and CD83 of Langerhans cells (LC) in the lesions of epidermodysplasia verruciformis (EV) patients.

METHODSWe used immunohistochemical method to detect the expressions of CD1a and CD83 in the lesions of 10 patients with EV lesions and in the skins of 10 normal subjects.

RESULTSNo CD83 + LCs was detected in all EV patients and normal controls, but CD1a + LC was found in all cases. The quantity of CD1a + LCs in the lesions of EV patients was significantly lower than that in the normal skin (P < 0.01); furthermore, the distribution of LCs in EV lesions was uneven.

CONCLUSIONThe functions of LCs may be inhibited in EV patients.

Antigens, CD ; biosynthesis ; genetics ; Antigens, CD1 ; biosynthesis ; genetics ; Epidermodysplasia Verruciformis ; immunology ; pathology ; Humans ; Langerhans Cells ; immunology ; Leukocyte Immunoglobulin-like Receptor B1 ; Receptors, Immunologic ; biosynthesis ; genetics ; Skin ; immunology ; pathology