Objective To evaluate the feasibility of perfusion weighted imaging (PWI) in the differentiation of recurrent glioma and radiation-induced brain injuries. Methods Fifteen patients with previously resected and irradiated glioma, presenting newly developed abnormal enhancement, were included in the study. The final diagnosis was determined either histologically or clinicoradiologically. PWI was obtained with a gradient echo echo-planar-imaging (GRE-EPI) sequence. The normalized rCBV ratio[CBV(abnormal enhancement)/CBV(contralateral tissue)], rCBF ratio[CBF(abnormal enhancement)/CBF(contralateral tissue)]and rMTT ratio[(MTT abnormal enhancement)/MTT(contralateral tissue)]were calculated, respectively. The regions of interest (ROIs) consisting of 20-40 mm2 were placed in the abnormal enhanced areas on postcontrast T1-weighted images. Ten to fifteen ROIs measurements were performed in each lesion and the mean value was obtained. Mann-Whitney test was used to determine whether there was a difference in the rCBV/rCBF/MTT ratios between glioma recurrence and radiated injuries. Results Nine of the 15 patients were proved recurrent glioma,6 were proved radiation-induced brain injuries. The mean rCBV ratio[2.87(0.70-4.91)]in glioma recurrence was markedly higher than that[0.70(0.12-1.62)]in radiation injuries (Z=-2.55,P＜0.05). The mean rCBF ratio[1.89(0.64-3.96)]in glioma recurrence was markedly higher than that[0.56(0.12-2.08)]in radiation injuries (Z=-2.08,P＜0.05). The areas under rCBV and rCBF ROC curve were 0.893 and 0.821. If the rCBV ratio ≤0.77, the diagnosis sensitivity of radiation-induced brain injuries was 100.0%;If ≥2.44, the diagnosis specificity of recurrent glioma was 100.0%. Conclusion PWI was an effective technique in distinguishing glioma recurrence from radiation injuries and rCBV and rCBF ratios were of great value in the differentiation.

OBJECTIVETo study the protective effects of Panax japonics (PJ) on alcohol-induced gastric lesion in mice and the possible mechanisms.

METHODMale ICR mice were randomized into six groups: normal, control, PJ (1.5, 3.0, 6.0 g x kg(-1)) and Yinduoan (1.5 g x kg(-1)). The mice were pretreated with PJ before administering ethanol to observe the effect on the concentration of ethanol in serum and urine. The contents of MDA, GSH and GSH-PX, CAT and SOD activities were measured in serum and gastric mucosa, and subsequently, the pathological evaluation of stomach was also observed.

RESULTThe concentration of ethanol in serum was evidently decreased after PJ (1.5, 3.0 g x kg(-1)) was administrated because the ethanol was eliminated fleetly through urine. Synchronously the PJ reduced the content of MDA and increased the GSH increased in serum and gastric, besides, it increased the enzymatic activities of GSHPX, CAT and SOD, and the ethanol-induced gastric mitochondria structure injury were ameliorated so as to make the function to normal.

CONCLUSIONBased on these observations, one could conclude that the PJ is a potent protective agent against ethanol-induced gastric damages. One mechanism may be related with inhibiting the absorbability of ethanol at gastrointestinal tract, decreasing the concentration of ethanol in serum, and accelerating the ethanol elimination through urine so as to alleviate the ethanol-induced damage to gastrointestinal mucosal, enhancing the first-pass metabolism in stomach, and particularly increasing the antioxidant levels in serum and gastric. These gastroprotective effects might be, at least partly, through ameliorating the gastric mitochondria structure.

Animals ; Catalase ; blood ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Ethanol ; Gastric Mucosa ; drug effects ; metabolism ; pathology ; Glutathione ; blood ; metabolism ; Glutathione Peroxidase ; blood ; metabolism ; Male ; Malondialdehyde ; blood ; metabolism ; Mice ; Mice, Inbred ICR ; Microscopy, Electron ; Mitochondria ; drug effects ; ultrastructure ; Panax ; chemistry ; Phytotherapy ; Plants, Medicinal ; chemistry ; Protective Agents ; isolation & purification ; pharmacology ; therapeutic use ; Random Allocation ; Stomach Diseases ; blood ; chemically induced ; prevention & control

OBJECTIVETo investigate the effects of tumor metastasis suppressor gene 1 (TMSG-1) overexpression on the proliferation, invasion and apoptosis of breast cancer cells and to determine possible correlations of TMSG-1 and metastasis of breast cancer.

METHODSFull-length human TMSG-1 coding sequences were cloned into plasmid pcDNA3.0-FLAG. The recombinant plasmids constructs were transfeced into MDA-MB-231, a highly malignant breast cancer cell line. Parental, vector-only stable transfectant and TMSG-1 stable transfectant clones were tested by MTT, soft agar colony formation and Boyden chamber assays. At twenty-four hours and forty-eight hours post transient transfection, double staining with Annexin-V-FITC and PI were employed to distinguish apoptotic cells from living cells by flow cytometry analysis.

RESULTSThree TMSG-1 overexpression clones were selected. Compared with the control cells, TMSG-1 overexpression MDA-MB-231 cells showed strong inhibition of proliferation and decreased clonogenicity in soft agar (P<0.05). Transfection of TMSG-1 into MDA-MB-231 cells significantly suppressed the cell invasion ability in vitro (decreased numbers of cells trespassing the matrigel in three experiments: 72.3+/-8.1, 85.0+/-4.2, and 73.5+/-7.8) in comparison with nave cells without transfection (187.5+/-2.1) and cells transfected with the control vector (162.3+/-6.8) (P<0.01). Transient transfection of TMSG-1 into MDA-MB-231 cells could promote cell apoptosis at 24 and 48 hours after transfection (P<0.05).

CONCLUSIONSTMSG-1 protein may have multiple functions in the regulation of proliferation, invasion and apoptosis of metastatic breast cancer cells, likely as a metastasis suppressor gene.

Apoptosis ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Membrane Proteins ; genetics ; metabolism ; physiology ; Neoplasm Invasiveness ; Plasmids ; Recombinant Proteins ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; physiology ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism ; physiology

OBJECTIVETo investigate the effects of tumor metastasis-related gene TMSG-1 overexpression on the proliferation and invasion of a highly metastatic prostate cancer cell line in vitro.

METHODSThe eukaryotic expression plasmids containing full-length TMSG-1 cDNAs were stably transfected into the highly metastatic prostate cancer cell line PC-3M-1E8. Clones highly expressing TMSG-1 were identified by RT-PCR and Western Blot analysis after G418 screening. The cell proliferation was detected by cell growth curve, MTT assay and soft agar colony formation assay. The invasive potential of tumor cells in vitro was tested by Matrigel invasion assay.

RESULTSThree TMSG-1 overexpression clones were selected. Cell growth curve and MTT assay showed that TMSG-1 overexpression clones exhibited a strong inhibition of proliferation compared with that of the parental cells or those transfected with vector alone from the third day of culture (P <0.05). In vitro analysis also showed that the TMSG-1 transfected clones exhibited a decreased clonogenicity in soft agar compared with that of the parental cells or those transfected with vector only (P < 0.05). TMSG-1 expression significantly suppressed cell invasion in vitro of TMSG-1-transfected PC-3M-IE8 cells (P < 0.05).

CONCLUSIONThe TMSG-1 protein may serve as a tumor metastasis suppressor due to inhibiting cell proliferation and invasion of the highly metastatic prostate cancer cell line PC-3M-1E8.

Animals ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; NIH 3T3 Cells ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Sphingosine N-Acyltransferase ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo investigate the regulatory mechanism of the transcription of tumor metastasis suppressor gene TMSG-1.

METHODSLuciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of TMSG-1. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were carried out to verify the interaction of KLF6 and Sp1 with the regulatory region of TMSG-1. Co-immunoprecipitation (CoIP) was performed to analyze the interaction between KLF6 and Sp1. TMSG-1 and wt-KLF6 mRNA expressions in cells with different metastatic capacities were quantitated by real-time PCR. Cell invasive capability was determined by Matrigel invasion assay.

RESULTSA 63 bp inducible regulatory region (+59 bp - +123 bp) in exon 1 was identified by luciferase assay using reporter plasmids with a series of TMSG-1 regulatory region deletions. Mutations in KLF6/Sp1 binding sites of this region resulted in a decrease of luciferase activity, while cotransfection with KLF6 or Sp1 expressing plasmids led to a remarkable increase of luciferase activity. EMSA and ChIP demonstrated that KLF6 as well as Sp1 interacted with this region. CoIP also indicated a possible interaction between KLF6 and Sp1 proteins. In the highly metastatic cell sublines, a low level of wild type KLF6 was associated synchronously with a low TMSG-1 level. Prostate carcinoma cells overexpressing KLF6 exhibited a higher TMSG-1 level and a lower invasive capability.

CONCLUSIONSTranscription factor complex of KLF6 and Sp1 may participate in the inducible transcriptional regulation of TMSG-1, and a decreased wild type KLF6 expression is likely associated with a low TMSG-1 level in the highly metastatic cell sublines.

Binding Sites ; genetics ; Cell Line, Tumor ; Electrophoretic Mobility Shift Assay ; Humans ; Immunoprecipitation ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutation ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Sp1 Transcription Factor ; genetics ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transcriptional Activation ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo observe the therapeutic efficacy of dietary boron supplement on retinoic acid-induced osteoporosis in rats, so as to provide experimental evidence for clinical management of osteoporosis with boron.

METHODSThirty-two SD rats were randomized into normal control group (8 rats) and osteoporotic group (24 rats), and osteoporosis was induced in rats of the latter group by intragastric retinoic acid administration at the daily dose of 80 mg/kg for 15 consecutive days. The osteoporotic rats were subdivided into control group (8 rats) without treatment, boron treatment group (8 rats) and estradiol treatment group (8 rats). After 30 days of treatment, the serum contents of Ca, P, boron and the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) in the rats were assayed, the bone mineral density (BMD) of the whole body, lumbar vertebrae and tibia were determined, and the morphological changes of the femurs were observed.

RESULTSThe serum contents of Ca and P in the rats of the 4 groups differed scarcely, but the content of boron in boron treatment group was markedly higher than that in the other three groups. In the osteoporotic control group, the activities of serum AKP and TRAP, the masses of spongy bone and cortical bone of the femurs, and the quantity of the osteoclasts were increased, with the BMD of the lumbar vertebrae and tibia decreased, suggesting osteoporotic conditions. The mean trabecular plate density and thickness, trabecular bone volume and cortical bone volume of the femurs in the osteoporotic rats treated with boron or estradiol were significantly increased, but the active osteoclast quantity in the spongy bone and serum TRAP activities were obviously decreased, and the bone quality was comparable with that of the normal group. In addition, the serum AKP activity and the active osteoblast quantity in the spongy bone were obviously increased in boron treatment group.

CONCLUSIONThe dietary boron supplement can increase the serum content of boron of osteoporotic rats to stimulate bone formation and inhibit bone resorption, producing therefore obvious therapeutical effect against osteoporosis.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Bone Density ; drug effects ; Boron ; administration & dosage ; therapeutic use ; Dietary Supplements ; Female ; Femur ; drug effects ; growth & development ; metabolism ; Isoenzymes ; blood ; Osteoporosis ; blood ; chemically induced ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase ; Time Factors ; Tretinoin

BACKGROUNDMycobacterium abscessus (M. abscessus) can cause a variety of human infections, involving the lung, skin and soft tissues, and is generally believed to be acquired from environmental sources. The aim of this study was to investigate the molecular diversity and antibiotic susceptibility of M. abscessus isolates as the basis for strategies to improve control and management of infection.

METHODSSeventy M. abscessus isolates from patients attending the Guangzhou Thoracic Hospital were identified from 2003 to 2005 by biochemical tests, gas chromatography, polymerase chain reaction (PCR)-restriction analysis (PRA) of heat shock protein gene hsp65, and sequencing of the quinolone resistance determining regions (QRDRs) of gyrA. Susceptibilities to six antibiotics were determined by micro-broth dilution. Isolates were genotyped using randomly amplified polymorphic DNA (RAPD) analysis.

RESULTSMost isolates (63/70; 90%) were susceptible to amikacin but rates of susceptibility to other antibiotics varied from moderate, clarithromycin (60%) and imipenem (43%), to low for ciprofloxacin and ofloxacin (3%), and 87% of isolates had intermediate susceptibility to cefoxitin. RAPD analysis showed that the 70 clinical isolates displayed 69 unique RAPD patterns.

CONCLUSIONSThe high genetic diversity of isolates suggests that they are not transmitted from person to person but, presumably, are acquired independently from environmental sources. M. abscessus isolates displayed variable levels of susceptibility to all antibiotics tested, other than amikacin, indicating a need for routine susceptibility testing to guide treatment.

Amikacin ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Cefoxitin ; pharmacology ; China ; Chromatography, Gas ; Ciprofloxacin ; pharmacology ; Clarithromycin ; pharmacology ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Mycobacterium ; drug effects ; genetics ; Polymerase Chain Reaction ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo study the expression and the location of vascular cell adhesion molecule-1 (VCAM-1) gene and its clinical significance in human oral squamous cell carcinoma (OSCC).

METHODSIn situ hybridization, PV-9000 polymer detection system for immunohistochemical staining was used to detect the expression and the location of VCAM-1 mRNA and VCAM-1 protein in 48 cases of OSCC and 10 cases of normal controls. Statistical analysis was performed using chi-square test in SPSS 13.0.

RESULTSVCAM-1 protein was mainly expressed in tumor cell cytoplasm and membrane, VCAM-1 mRNA was mainly expressed in tumor cell cytoplasm. The expression rate of VCAM-1 mRNA and VCAM-1 protein was significantly higher in OSCC than that in normal oral mucosa (P<0.01). The expression of VCAM-1 mRNA was positively correlated with that of VCAM-1 protein (P<0.01). In the clinicopathologic factors, lymph node metastasis and depth of infiltration were closely correlated with VCAM-1 expression (P<0.01). The expression of VCAM-1 was significantly higher in tumor with lymph node metastasis than in tumor without lymph node metastasis (P<0.01).

CONCLUSIONOverexpression of VCAM-1 gene in OSCC may play a potential role in the development of OSCC. The overexpression of VCAM-1 gene in OSCC may be related to the tumor infiltration and metastasis.

Carcinoma, Squamous Cell ; Humans ; In Situ Hybridization ; Lymphatic Metastasis ; Middle Aged ; Mouth Mucosa ; Mouth Neoplasms ; RNA, Messenger ; Vascular Cell Adhesion Molecule-1

OBJECTIVETo investigate the correlation between matrix metalloproteinase 9 (MMP-9) expression and tumor invasion and metastasis as well, and to explore the potential application of controlled expression of target gene in tumor gene therapy.

METHODSOne self-contained tetracycline-regulated retroviral vector containing anti-sense cDNA of MMP-9 was constructed and transfected into a metastatic human melanoma cell line WM451 which expressed a high level of MMP-9. Techniques such as growth rate measurment, MTT assay, (3)H-thymidine incorporation, colony forming ability in soft agar, invasion assay in Boyden chamber, as well as zymography and Western blot were applied to analyze the expression of MMPs and behaviors of tumor cells in vitro before and after gene transfection. Tumorigenecity and spontaneous metastasis were tested in nude mice.

RESULTSIn the presence of exogenous tetracycline, the transfected antisense MMP-9 did not affect the endogenous level of MMP-9 in WM451 cells, and showed no significant changes in cell behaviors in comparison with that of the vector-transfected control cells. Nevertheless, withdrawal of tetracycline from the medium caused a significant down-regulation of expression and activity of MMP-9. The capacity of cell growth in vitro, colony forming ability in soft agar, invasion through Matrigel all were inhibited remarkably when compared with the controls. Spontaneous metastasis in nude mice was significantly inhibited.

CONCLUSIONSTransfection of anti-sense MMP-9 can down-regulate the invasion and metastasis of melanoma cells both in vitro and in vivo, further clarifying the important role of MMP-9 in tumor progression.

Animals ; Cell Division ; Cell Line, Tumor ; DNA, Antisense ; DNA, Complementary ; genetics ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Genetic Vectors ; Humans ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Melanoma ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Retroviridae ; genetics ; Tetracycline ; pharmacology ; Transfection

OBJECTIVETo determine the role of extracellular signal-regulated kinase (ERK1/2) and p38 cascades in P2Y receptor-evoked effects on prostatic cancer cells.

METHODSHighly metastatic prostatic cancer cells 1E8 were transfected with dominant-negative MAPK kinase 1 (KA-MEK1). The activation of ERK1/2 was determined by Western blot technique. The role of ERK1/2 and p38 cascades in P2Y receptor-evoked effects on in vitro growth, colony formation and in vitro invasion was detected by cell count, soft agar colony formation assay and in vitro invasion assay. The effect of ATP on apoptosis was detected by flow cytometry.

RESULTSERK1/2 activity in 1E8-KA-MEK1 transfectants was significantly suppressed by dominant-negative MEK1 transfection. After culture of 6 days, 1E8-KA-MEK1 transfectants exhibited a growth inhibition of 71% as compared with 1E8-pcDNA3 control. Moreover, after continuous treatment with 100 micro mol/L ATP for 6 days, the growth of 1E8-KA-MEK1 transfectants was further inhibited by an additional 17.2%. Pretreatment with 10 micro mol/L p38 inhibitor SB203580 antagonized the effect of ATP-induced additional growth inhibition, suggesting that ERK1/2 and p38 pathways play an important role in ATP-induced growth inhibition. In soft agar assay, 1E8-KA-MEK1 transfectants formed smaller colonies and exhibited a 75% decrease in colony formation (as compared with control). Further treatment with ATP or SB203580 plus ATP did not show significant effect on colony formation of 1E8-KA-MEK1 cells, implying a potential role of ERK1/2, instead of p38, in P2Y receptor-mediated inhibitory effect on colony formation. In in vitro invasion assay, 1E8-KA-MEK1 cells showed a 41% decrease in passing through matrigel-coated membranes, as compared with control. Treatment with ATP could restore their invasive ability, and this effect by ATP could be blocked by pretreatment with SB203580, indicating the involvement of both ERK1/2 and p38 pathways in invasive ability of prostatic cancer cells.

CONCLUSIONSThe effects of ATP on in vitro growth, invasion and colony formation of prostatic cancer cells depend on the status of P2Y receptor activation by different treatment protocols. Continuous activation of P2Y receptor results in growth inhibition and transient activation of P2Y receptor stimulates in vitro invasion of prostatic cancer cells. Both ERK1/2 and p38 pathways are responsible for these effects; but only the ERK1/2 pathway is involved in regulation of colony formation of prostatic cancer cells.

Adenosine Triphosphate ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Imidazoles ; pharmacology ; MAP Kinase Kinase 1 ; metabolism ; Male ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Neoplasm Invasiveness ; Prostatic Neoplasms ; enzymology ; pathology ; Pyridines ; pharmacology ; Receptors, Purinergic P2 ; metabolism ; physiology ; Transfection ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors