A large amount of nicotinamide adenine dinucleotide phosphate (NADPH) is required for fatty acid synthesis and maintenance of the redox state in cancer cells. Malic enzyme 1(ME1)-dependent NADPH production is one of the three pathways that contribute to the formation of the cytosolic NADPH pool. ME1 is generally considered to be overexpressed in cancer cells to meet the high demand for increased de novo fatty acid synthesis. In the present study, we found that glucose induced higher ME1 activity and that repressing ME1 had a profound impact on glucose metabolism of nasopharyngeal carcinoma(NPC) cells. High incorporation of glucose and an enhancement of the pentose phosphate pathway were observed in ME1-repressed cells. However, there were no obvious changes in the other two pathways for glucose metabolism: glycolysis and oxidative phosphorylation. Interestingly, NADPH was decreased under low-glucose condition in ME1-repressed cells relative to wild-type cells, whereas no significant difference was observed under high-glucose condition. ME1-repressed cells had significantly decreased tolerance to low-glucose condition. Moreover, NADPH produced by ME1 was not only important for fatty acid synthesis but also essential for maintenance of the intracellular redox state and the protection of cells from oxidative stress. Furthermore, diminished migration and invasion were observed in ME1-repressed cells due to a reduced level of Snail protein. Collectively, these results suggest an essential role for ME1 in the production of cytosolic NADPH and maintenance of migratory and invasive abilities of NPC cells.
Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Survival ; Glucose ; metabolism ; Glycolysis ; Humans ; Malate Dehydrogenase ; metabolism ; NADP ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Oxidation-Reduction ; Oxidative Phosphorylation ; Pentose Phosphate Pathway ; Proto-Oncogene Proteins c-akt ; metabolism

AIM:To evaluate the effect of α2-macroglobulin(α2M) against X-ray induced obstacle on osteo-genic differentiation of human bone marrow mesenchymal stem cells(hBMMSCs). METHODS:hBMMSCs were cultured in vitro. The 4th generation of hBMMSCs was irradiated with 8 Gy X-ray,then induced osteogenic differentiation and trea-ted with different concentrations of α2M(0.5 and 1.0 g/L). The alkaline phosphatase(ALP) activity and the mRNA ex-pression of runt-related transcription factor-2 (RUNX2) were detected on day 7 after osteogenic induction. The protein ex-pression of osteoglycin (OGN) was evaluated by Western blot on day 14 after osteogenic induction. The formation of calci-um nodules was detected by alizarin red staining on day 21 after osteogenic induction. The activity of superoxide dismutase (SOD) and the protein expression of MnSOD of irradiated hBMMSCs with 8 Gy X-ray were determined at 24 h after α2M treatment. RESULTS:Compared with 8 Gy X-ray group,the activity of ALP,the mRNA expression of RUNX2,the pro-tein expression of OGN and MnSOD,as well as SOD activity were higher than those in the hBMMSCs treated with α2M at 0.5 and 1.0 g/L after 8 Gy X-ray irradiation,and the calcium nodules were also increased. CONCLUSION:α2M signifi-cantly improves the osteogenic differentiation ability,the SOD activity and MnSOD protein expression of hBMMSCs after ra-diation injury.

OBJECTIVETo measure breast basic dimension by using computer-aided projection fringe system.

METHODSA system has been developed for measuring breast basic dimension based on computer-aided projection fringe measurement and programming software. Plastic manikins breast's SN-N (sternal notch to nipple distance), N-ML (nipple to midline distance), N-N (internipple distance), MBW (base width of breast) and N-IMF (nipple to inframammary fold distance) are measured with this system. At the same time, these items are also measured with routine ruler.

RESULTSThis study indicate that the system has some merits: (1) non-touching measurement; (2) it is very rapid, the patient measured need hold his breath only 0.5 second, and all the time it takes is about 2.5 minutes; (3) the measurement's sensitivity is as high as to 0.6 mm, which meets the clinic requirement entirely; (4) the measurement's accuracy of the system is not significantly when comparing to the routine ruler's.

CONCLUSIONSComputer-aided projection fringe system for measuring breast basic dimension is feasible and advanced.

Breast ; anatomy & histology ; Female ; Humans ; Image Processing, Computer-Assisted ; Tomography, Optical ; methods

OBJECTIVETo study the chemical constituents contained in ethanol extracts from aerial parts of Emilia sonchifolia.

METHODThe compounds were separated and purified with various chromatographic techniques, and their structures were identified on the basis of physicochemical properties and spectral data.

RESULTFifteen compounds were separated from ethyl acetate fraction of 90% ethanolic extract and identified as rhamnetin (1), isorhamnetin (2), quercetin (3), luteolin (4), tricin-7-O-beta-D-glucopyranoside (5), 8-(2"-pyrrolidinone-5"-yl) -quercetin (6), 5, -2', 6'-trihydroxy-7, 8-dimethoxyflavone-2'-O-beta-D-glucopyranoside (7), succinic acid (8), fumaric acid (9), p-hydroxybenzoic acid (10), 4-hydroxy isophthalic acid (11), 3, 4-dihydroxycinnamic acid (12), esculetin (13), isowedelolactone (14) and uracil (15), respectively.

CONCLUSIONAll compounds except compound 3 were separated from this genus for the first time.

Asteraceae ; chemistry ; Plant Extracts ; analysis

OBJECTIVETo investigate the differential expression of toll-like receptor 2 (TLR2), toll-like receptor 4 (TLR4) and their potential role in the pathogenesis of chronic suppurative otitis media and cholesteatoma.

METHODSNormal canal skin of 30 patients with tympanosclerosis were enrolled as control, 30 cases with chronic suppurative otitis media and 30 patients with cholesteatoma were studied. Real-time PCR, Western blot and Immunohistochemistry were preformed to detect the expression of TLR2/TLR4 in normal canal skin, mucosa and granulation tissue of chronic suppurative otitis media, mucosa, granulation tissue, cholesteatoma epithelium of cholesteatoma, and the differential expression were analyzed.

RESULTS(1) the mRNA and protein expression of TLR2 and TLR4 were detected in all normal canal skin, mucosa and granulation tissue of chronic suppurative otitis media, mucosa, granulation tissue, cholesteatoma epithelium of cholesteatoma. (2) Both mRNA and protein level of TLR2/TLR4 in mucosa of chronic suppurative otitis media and cholesteatoma were higher than those in normal canal skin, but lower in cholesteatoma epithelium, there was no significant difference in mucosa of the two otitis media groups. (3) The mRNA and protein expression of TLR2/TLR4 in granulation tissue of chronic suppurative otitis media and cholesteatoma were significant increased when compared with normal canal skin, and TLR2 expression level was higher in granulation tissue of cholesteatoma than in chronic suppurative otitis media. (4) TLR2/TLR4 positive cells mainly infiltrated in granulations, significantly more than in normal skin, while fewer in the epithelium of cholesteatoma.

CONCLUSIONSDifferential expression of TLR2 and TLR4 in mucosa suggests middle ear is a TLR2/TLR4 participated functional modulation of the innate immune system and also suggests that they may play a different role in the pathophysiology of chronic otitis media and cholesteatoma.

Case-Control Studies ; Cholesteatoma, Middle Ear ; metabolism ; Chronic Disease ; Humans ; Otitis Media, Suppurative ; metabolism ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo look for a gene delivery route to the treatment of Duchenne muscular dystrophy(DMD).

METHODSThe recombinant adeno-associated virus vector(rAAV) carrying a LacZ reporter gene was constructed. rAAVLacZ was delivered into the skeletal muscle tissue of C57/BL6 mice by intramuscular injection. Then an intraarterial delivery route was taken to reveal whether rAAVLacZ could transduce muscle tissue.

RESULTS(1) The LacZ gene was efficiently transduced and expressed persisting for 5 months after intramuscular injection. (2) The membrane of muscle and smooth muscle of vessel was widely transduced by intra-arterial delivery rAAVLacZ.

CONCLUSIONThese data provide the evidence that rAAVLacZ can efficiently transduce muscle for a long period. Improving intraarterial gene delivery will be promising means for rAAV-mediated gene therapy for generalized skeletal muscle of DMD.

Animals ; Cell Line ; Dependovirus ; genetics ; Embryo, Mammalian ; Gene Expression ; Genes, Reporter ; Genetic Vectors ; Humans ; Injections, Intramuscular ; Kidney ; cytology ; metabolism ; Lac Operon ; physiology ; Mice ; Mice, Inbred C57BL ; Muscle, Skeletal ; cytology ; metabolism ; Recombinant Proteins ; metabolism ; Transfection ; methods

OBJECTIVEQuantitative study of the effect of anti-human VEGF mAb E11 to VEGF level in serum of nude mice transplanted buccal carcinoma.

METHODSE11 was administered into BALB/c nu/nu mice which were transplanted human buccal carcinoma. The saline was administrated as negative control. Mice were killed at 18 days. The VEGF level in serum of mice was determined by improved indirect ELISA.

RESULTSCompared with the VEGF level in serum of mice in saline group, it was dramatically decreased in E11 group. The VEGF level in serum of mice treated E11 by subcutaneous was lowest and only reached (1.17 +/- 0.13) microg/L.

CONCLUSIONIt demonstrated that the anti-human VEGF mAb could reduce the VEGF level in serum by binding VEGF, and block its biological activity. It indicates that VEGF in serum of malignant tumor patient is a new tumor marker.

Animals ; Antibodies, Monoclonal ; pharmacology ; Biomarkers, Tumor ; blood ; Carcinoma ; blood ; Humans ; Mice, Inbred BALB C ; Mice, Nude ; Mouth Mucosa ; pathology ; Neoplasm Transplantation ; Neoplasms ; blood ; Vascular Endothelial Growth Factor A ; blood

[Objective]To explore the effect of alpha2-macroglobulin(α2M)on superoxide anion(.O2-)content, superoxide dismutase(SOD)activity and the process of cell-to-myofibroblast transformation in human skin fibroblasts (HSF)after X-ray irradiation.[Methods]HSF cells were irradiated with 0,5,10,15 and 20 Gy X-ray.The change of. O2- content and SOD activity in the supernatant of cell culture medium were measured on the first day after irradiation. The protein expression of alpha-smooth muscle actin(α-SMA)was detected by Western blot on the fifth day after irradia-tion.The most sensitive radiation dose is selected.HSF cells were irradiated with the above sensitive dose.Respectively, 1h before irradiation,1 h after irradiation,the experimental group cell culture medium was added to a final concentration of 0.25 mg/mL,0.5 mg/mL of α2M.The change of.O2-content,SOD activity and the protein expression of α-SMA were detected.[Results]HSF cells were irradiated with 5~20 Gy doses of X-ray..O2- content increased,SOD activity de-creased and α-SMA protein expression increased gradually(P<0.05).The addition of α2M at 1 h after 10Gy X-ray irradi-ation reduced the.O2- content,increased the SOD activity and downregulated the protein expression of α-SMA in HSF cells(P<0.05). There was no significant change in the administration at 1 h before irradiation.[Conclusion]HSF cells increased.O2-content significantly,while SOD activity decreased,and the tendency to transform myofibroblasts after X-ray irradiation.α2M can reduce the.O2-content,increase the SOD activity in HSF cells and inhibit the transformation of fibroblasts into myofibroblasts after irradiation.Indicating that α2M can play a role in radiation protection by anti-oxida-tion and anti-fibrosis.

OBJECTIVETo assess the feasibility of the 10 microg recombination yeast hepatitis B vaccine in the expanded applicable population group aged 5 - 18.

METHODSPeople with both HBsAg and anti-HBs negative were selected to take two-stage clinical experiment and the safety and immunogenicity were observed. Safety observation was conducted in 925 subjects, while 568 for immunogenicity. The observation group (aged 5 - 18) included 493 subjects, and (age > 18) 75 enrolled in control group. For the observation group, there were three sub-groups including a child group (141, aged 5 - 6), early youth group (177, aged 12 - 13), and youth group (175, aged 16 - 18). Both groups were administered with 10 microg recombination yeast hepatitis B vaccines with 3 doses at 0 month, 1st month, 6th month. To assess the immunogenicity, the vaccination reactions were observed during the following 4 weeks in order to assess the vaccine safety. The blood samples were taken during 4 - 6 weeks after fully vaccinated, and then anti-HBs were tested with RIA and analyzed by comparing the positive rate of anti-HBs, the geometric mean titer (GMT) and the protective rate between the two groups.

RESULTSBoth observation and control group didn't show any general reactions, adverse events following immunization (AEFI) or coincidental cases when observed at 0.5 h, 6 h, 24 h, 48 h, 72 h, 1 week, 2 weeks, 3 weeks, 4 weeks after being vaccinated. The result of serum test showed, the positive rates of child group, early youth group, youth group and control group were respectively 100.00% (141/141), 97.18% (172/177), 98.29% (172/175) and 89.33% (67/75); the GMTs of anti-HBs were respectively 440.28, 875.38, 467.80, 131.06 U/L; the protective rates were respectively 100.00% (141/141), 97.18% (172/177), 97.14% (170/175) and 86.67% (65/75). The positive rate, GMT and protective rate of the experimental group were all higher than that of control group (chi(2)(positive rate) = 12.77, 5.12, 7.99; t(GMT) = 3.89, 4.13, 5.91; chi(2)(protective rate) = 16.81, 8.60, 8.44; P < 0.05).

CONCLUSIONThis vaccine could be expanded to 5 - 18 year-old population with safety and effectiveness, the positive rate and protective rate of anti-HBs were both higher than that of control group.

Adolescent ; Child ; Child, Preschool ; Female ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B Vaccines ; administration & dosage ; adverse effects ; immunology ; Humans ; Male ; Vaccines, Synthetic ; administration & dosage ; adverse effects ; immunology

【Objective】To establish a cell model of radiation injury of human bone marrow mesenchymal stem cells（hBMMSC），and to provide experimental basis for further study on the pathogenesis of osteoradionecrosis of the jaws（ORNJ）.【Methods】After X-ray irradiation，the gene expression change of Beclin1，Sox2，Nanog，RUNX2 and OGN in hBMMSC were evaluated by Western blot and RT-qPCR. The apoptosis rate change was detected by Annexin- V/PI double staining method. The proliferation rate change was determined by clone formation experiment. The alkaline phosphatase activity change was detected by microplate method.【Results】The gene expressions of Beclin1 increased with the doses increased，while Sox2，Nanog，RUNX2 and OGN in hBMMSC were all down- regulated after irradiation. The apoptotic rate increased，the colony formation rate decreased and the alkaline phosphatase activity decreased also.【Conclusion】X-ray irradiation can cause ionizing radiation injury of hBMMSC. We successfully established a cell model of hBMMSC radiation injury，the model can be applied to the experimental study of the pathogenesis of ORNJ.