Objective To construct a lentiviral vector containing mir-7-3 gene and green fluorescent protein (GFP) gene,and to detect the expression of mir-7-3 gene in U251 cells.Methods The fragments containing all the mir-7-3 gene were amplified by RT-PCR and were cloned into the lentivirus vectors labeled with GFP,which was transfected together with the packaging plasmids into 293T cells by CaC12.The supernatant was collected,concentrated,identified,and was transfected to U251 cells of gliomas.Fluorescent microscopy was used to observe the fluorescence in the 293T cell,and real time RT-PCR was used to examine the relative contents of mir-7-3 in U251 cells.Results Electrophores was shown that the sequence of the RT-PCR product was consistent with the data of mir-7-3 by DNA sequence analysis,indicating that the mir-7-3 gene was successfully cloned,and strong green fluorescence was observed by fluorescent microscopy.The supernatant of lentivirus-transfected 293T cells effectively infected U251 cells and the relative content of mir-7-3 was observed in the transfected U251 cells.Conclusion It is concluded that the lentiviral vector containing mir-7-3 gene was constructed successfully,which provides a basis for further study of mir-7-3 function.

BACKGROUND:Macrophage migration inhibitory factor is involved in the process of a variety of diseases, and plays a very important role in the tumor, autoimmune diseases, inflammation, angiogenesis, fibrotic diseases and so on. These biological characteristics are similar to keloids. OBJECTIVE: To compare the distribution and number of macrophage migration inhibitory factor in normal skin, hypertrophic scar and keloid. METHODS: We colected 40 clinical pathological scar specimens after surgery, including 20 hypertrophic scars and 20 keloids. Another 10 samples of the normal skin were used as control group. Hematoxylin-eosin staining and immunohistochemistry staining were performed to test the expression of macrophage migration inhibitory factor in pathological scars and normal skin. RESULTS AND CONCLUSION:Macrophage migration inhibitory factor was positively expressed in the normal skin, hypertrophic scar and keloid, and the expression of macrophage migration inhibitory factor in keloid was significantly higher than that in hypertrophic scar and normal skin (P < 0.01). It means that the abnormal infiltration of macrophage migration inhibitory factor may be associated with the formation of keloid.

OBJECTIVETo evaluate whether garlicin can prevent reperfusion no-reflow in a catheter-based porcine model of acute myocardial infarction (AMI).

METHODSTwenty-two male Chinese mini swines were randomized into 3 groups: sham-operation group (n=6), control group (n=8), and garlicin group (n=8). The distal part of left anterior descending coronary artery (LAD) in swines of the latter two groups was completely occluded by dilated balloon for 2 h and a successful AMI model was confirmed by coronary angiography (CAG) and electrocardiograph (ECG), which was then reperfused for 3 h. In the sham-operation group, balloon was placed in LAD without dilatation. Garlicin at a dosage of 1.88 mg/kg was injected 10 min before LAD occlusion until reperfusion for 1 h in the garlicin group. To assess serial cardiac function, hemodynamic data were examined by catheter method before AMI, 2 h after occlusion and 1, 2, and 3 h after reperfusion. Myocardial contrast echocardiography (MCE) and double staining with Evans blue and thioflavin-S were performed to evaluate myocardial no-reflow area (NRA) and risk area (RA).

RESULTSLeft ventricular systolic pressure and left ventricular end-diastolic pressure significantly improved in the garlicin group after reperfusion compared with the control group P<0.05) and 2 h after AMI (P<0.05). MCE showed garlicin decreased reperfusion NRA after AMI compared with the control group (P <0.05). In double staining, NRA/RA in the garlicin group was 18.78%, significantly lower than that of the control group (49.84%, P<0.01).

CONCLUSIONSGarlicin has a preventive effect on the porcine model of myocardial infarction reperfusion no-reflow by improving hemodynamics and decreasing NRA.

Allyl Compounds ; pharmacology ; therapeutic use ; Animals ; Cardiotonic Agents ; pharmacology ; therapeutic use ; Contrast Media ; Disease Models, Animal ; Disulfides ; pharmacology ; therapeutic use ; Hemodynamics ; drug effects ; Male ; Myocardial Infarction ; complications ; diagnostic imaging ; drug therapy ; pathology ; Myocardial Reperfusion ; No-Reflow Phenomenon ; complications ; diagnostic imaging ; drug therapy ; pathology ; Swine ; Swine, Miniature ; Thiazoles ; metabolism ; Ultrasonography

BACKGROUND: Few studies investigated serum uric acid levels in patients with acute ST-elevation myocardial infarction (STEMI). The study was to assess the clinical value of serum uric acid levels in patients with acute ST-elevation myocardial infarction (STEMI). METHODS: Totally 502 consecutive patients with STEMI were retrospectively studied from January 2005 to December 2010. The level of serum lipid, echocardiographic data and in-hospital major adverse cardiovascular events (MACE) in patients with hyperuricemia (n=119) were compared with those in patients without hyperuricemia (n=383). The relationship between the level of serum uric acid and the degree of diseased coronary artery was analyzed. All data were analyzed with SPSS version 17.0 software for Student's t test, the Chi-square test and Pearson's correlation coefficient analysis. RESULTS: Serum uric acid level was positively correlated with serum triglyceride level. Hyperlipidemia was more common in hyperuricemia patients than in non-hyperuricemia patients (43.7％ vs. 33.7％, P=0.047), and serum triglyceride level was significantly higher in hyperuricemia patients (2.11±1.24 vs. 1.78±1.38, P=0.014). But no significant association was observed between serum uric acid level and one or more diseased vessels (P>0.05). Left ventricular end-diastolic diameter (LVEDd) was larger in hyperuricemia patients than in non-hyperuricemia patients (53.52±6.19 vs. 52.18±4.89, P=0.041). The higher rate of left systolic dysfunction and diastolic dysfunction was discovered in hyperuricemia patients (36.4％ vs. 15.1％, P<0.001; 68.2％ vs. 55.8％, P=0.023). Also, hyperuricemia patients were more likely to have in-hospital MACE (P<0.05). CONCLUSIONS: Serum uric acid level is positively correlated with serum triglyceride level, but not with the severity of coronary artery disease. Hyperuricemia patients with STEMI tend to have a higher rate of left systolic dysfunction and diastolic dysfunction and more likely to have more in-hospital MACE.

OBJECTIVETo establish a novel Myc gene transgenic mouse model for spontaneously forming B-lymphoma and assessing its tumorigenesis potential.

METHODSFreshly isolated hematopoietic progenitor cells served as the target for Myc gene transfer mediated by a retrovirus vector. These cells were engrafted into C57BL/6 mice with (60)Co-gamma ray radiation in advance. Tumor latency was measured and the tumor loaded mice were followed for survival time. Tumor was identified with histology and immunostaining. The exogenous Myc gene was detected by Western blot (in liver, spleen, tumor tissue) and flow cytometry (FCM) \[in bone marrow (BM)\].

RESULTSMice BM-infected with mutant Myc gene more readily gave rise to B-cell lymphomas than those infected with wild type Myc gene did Myc gene was expressed highly in BM and tumor tissues but not in liver and spleen.

CONCLUSIONOur model will be a tool in assessing the transforming potential of Myc mutants and in studying cooperation between Myc and other oncogenes. Mutant Myc is more effective than wild-type Myc in promoting B cell lymphomagenesis in mice.

Animals ; B-Lymphocytes ; Cell Transformation, Neoplastic ; Flow Cytometry ; Lymphoma ; Lymphoma, B-Cell ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Retroviridae Infections

ObjectiveTo investigate the CT and MRI characteristic features of neuroendocrine carcinoma in paranasal sinuses.MethodsCT and MRI findings of 10 patients with proved neuroendocrine carcinoma by pathology were retrospectively reviewed. All patients underwent plain and enhanced MRI scanning,and 9 patients also underwent CT manning.ResultsThere were 5 males and 5 females with mean age of (48 ± 9 ) years old,ranging from 27 to 57 years.The treatment time after symptoms onset ranged from 1 to 4 months,with the median of 2 months.Clinical symptoms were headache and vision loss,hyposmia and yellow nasal discharge,and exophthalmos.The lesions were located in the ethmoidal sinus ( n =6 ),maxillary sinus ( n =2),and bilateral sphenoid sinus ( n =5 ).The lesions were symmetrical in the sphenoid sinus.Pathology type included typical carcinoid tumor ( n =1 ),atypical carcinoid ( n =1 ),and neuroendocrine carcinoma not otherwise specified ( n =8 ). Immunohistochemical staining showed that neurospecific enolase,synaptophysin,cytokeratin and P53 were all positive.On CT images,lesions showed isointensity (n =1 ),iso- to hypointense (n =4 ),and iso- to hyperintense (n =4 ) with hypointense or hyperintense spots.Bone changes included bony absorption and sclerosis ( n =1 ) with a clear margin in typical carcinoid tumor,and moth-eaten bone destruction in other 8 cases( n =8).The lesions were isointense on T1-weighted images,and isointense (n =4) or mixed iso- to hyperintense on T2-weighted images (n =6).Lesions showed mild to medium heterogeneous enhancement ( n =7 ) or marked enhancement ( n =3 )on gadolinium-enhanced images.Time-signal intensity curve ( TIC ) showed plateau type in 2 cases.The aggressive nature of the tumors was demonstrated by invasion of adjacent structures,involvement of nasal cavity( n =9 ),orbits ( n =7 ),pterygopalatine fossa ( n =4 ),ethmoidalsinus and sphenoid ( n =3 ),clivus ossis occipitalis(n =2),cavernous sinus and internal carotid canal(n =2),optic canal(n =2),jugular fossa ( n =1 ),anterior fossa ( n =1 ),apex partis petrosae ossis temporalis ( n =1 ),meninges ( n =1 ),temporal fossa and infratemporal fossa ( n =1 ),pharyngonasal cavity and parapharyngeal space ( n =1 ).ConclusionsThere are different CT features in different pathological types of neuroendocrine carcinoma of the paranasal sinuses,and MRI can demonstrate the invasive extent accurately. CT combined MRI can provide more comprehensive information in the diagnosis and therapy.

Objective To evaluate the incidence of postoperative hemorrahage in children undergoing adenoidectomy , and to discuss its possible causes. Methods Included in this study were children who underwent adenoid and/or tonsil surgery at Shenzhen Chilidren's Hospital between January 2004 and November 2009. The change of hemoglobin ( Hb) and hematocrit ( Hct) were retrospectively analysed. The blood loss was estimated by the change of Hct. Results There were 2078 cases that accomplished the inclusion criteria in the period of study. Ten children bled 0. 5 - 4. 0 hours after surgery,without superfluous hemorrahage during the operation and post-tonsillectomy. This represented an incidence of 0.48% of immediate postoperative haemorrhage among the 2078 procedures analyzed. Statistical differences were found between boys (0.21%) and girls (1.10%, x2 =5.597, P<0.05). The change of Hb and Hct was positively correlated (r = 0. 95 , P < 0. 01) , the blood loss was positively correlated with the bleeding time ( r = 0. 66, P < 0. 05 ) . The causes of postoperative hemorrhage were coagulation system deficits, chronic nasopharyngitis, deficient hemostasis and immoderate ravage. To control the postoperative hemorrhage, 2 postnasal packing under topical anaesthesia and 8 electrocautery under general anaesthesia were applied. Conclusions Poor operative technique and deficient hemostasis are the major causes of primary hemorrahage. Prompt operation to control the postoperative bleeding should be done 2 hours after bleeding under general anesthesia in order to avoid severe complications.

OBJECTIVETo study how hepatitis B virus(HBV) 'a' determinant hotpoint mutations were influecing the hepatitis B vaccine efficacy.

METHODSPrimers were designed in HBV conservative region, and the degenerate probes for detecting 16 'a' determinant hotpoint mutations were developed for gene chips. Sensitivity and specificity of the gene chips were evaluated by clone sequencing. Sera of 47 pairs of mothers and infants with immune failure and 323 mothers of children with immune protection of HB vaccine were detected by the gene chips.

RESULTSResult from clone sequencing demonstrated that the gene chips were specific for the detection of 'a' determinant hotpoint mutations. The wild type of HBV was still dominant, with the prevalence of 78.66%, and the mutation frequencies of 126A, 145R, 126S-1, 126S-2, 129H, 144A, and 129R were 11.27%, 5.76%, 5.28%, 4.56%, 1.20%, 0.72% and 0.24%, respectively. The prevalence of 126A mutation was significantly higher than that of other mutations(P < 0.01). No significant differences were found in mother-infant transmission rates of 126A, 126S-1, 126S-2 and 145R variants.

CONCLUSIONThe currently available hepatitis B vaccine could block mother-infant transmission of 126A, 126S and 145R variants. It appears that there is no need to develop a new hepatitis B vaccine against 126 and 145 variants at present, but the consistent epidemiological surveillance on HBV mutants should be carried out.

Adult ; Female ; Genotype ; Hepatitis B ; prevention & control ; transmission ; Hepatitis B Vaccines ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control ; Mutation ; Oligonucleotide Array Sequence Analysis ; Pregnancy ; Pregnancy Complications, Infectious ; prevention & control ; virology

OBJECTIVETo investigate the biological functions of TTG1A in liver fibrosis.

METHODSYeast two-hybrid system was used to screen proteins associated with TTG1A. Briefly, the coding sequence of TTG1A was cloned into pGBKT7 vector, and the recombinant plasmid was transformed into yeast cells AH109 ( a type), then these cells were mated with yeast cells Y187 (a type) transformed with human leukocyte cDNA library plasmid pACT2. The obtained diploid yeast cells were plated on synthetic dropout nutrient medium containing X-alpha-gal for double selection. The plasmids from positive colonies were transformed into E.coli and sequenced.

RESULTSThe recombinant yeast expression vector pGBKT7-TTG1A was successfully constructed. Nineteen TTG1A binding proteins, including Homo sapiens major histocompatibility complex, class II DP beta 1 (HLA-DPb1), Homo sapiens ribosomal protein L30 (RPL30), Homo sapiens nucleophosmin Homo sapiens nucleobindin 2 (NUCB2), Homo sapiens ash2, variant Gaucher disease and variant metachromatic leukodystrophy, MORF4L1, Homo sapiens ubiquitin-conjugating enzyme E2L3 (UBE2L3), APOA1, Homo sapiens lectin, and galectin 1, were identified.

CONCLUSIONSThis study may help to elucidate the molecular function of TTG1A.

Carrier Proteins ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Library ; Genes, Regulator ; Genetic Vectors ; Hepatic Stellate Cells ; Humans ; Liver Cirrhosis ; genetics ; Oligonucleotide Array Sequence Analysis ; Plasmids ; genetics ; Ribosomal Proteins ; genetics ; Transcriptional Activation ; Transforming Growth Factor beta1 ; genetics ; Two-Hybrid System Techniques ; Yeasts ; genetics

OBJECTIVETo investigate the role of monocarboxylate transporter 1 (MCT1) in enhancing the sensitivity of breast cancer cells to 3-bromopyruvate (3-BrPA).

METHODSThe inhibitory effect of 3-BrPA on the proliferation of breast cancer cells was assessed with MTT assay, and brominated propidium bromide single staining flow cytometry was used for detecting the cell apoptosis. An ELISA kit was used to detect the intracellular levels of hexokinase II, lactate dehydrogenase, lactate, and adenosine triphosphate, and Western blotting was performed to detect the expression of MCT1. MDA-MB-231 cells were transiently transfected with MCT1 cDNA for over-expressing MCT1, and the effect of 3-BrPA on the cell proliferation and adenosine triphosphate level was deteced.

RESULTS3-BrPA did not produce significant effects on the proliferation and apoptosis of MDA-MB-231 cells, and the cells treated with 200 µmol/L 3-BrPA for 24 h showed an inhibition rate and an apoptosis rate of only 8.72% and 7.8%, respectively. The same treatment, however, produced an inhibition rate and an apoptosis rate of 84.6% and 82.3% in MCF-7 cells, respectively. In MDA-MB-231 cells with MCT1 overexpression, 200 µmol/L 3-BrPA resulted in an inhibition rate of 72.44%, significantly higher than that in the control cells (P<0.05); treatment of the cells with 25, 50, 100, and 200 µmol/L 3-BrPA for 6 h resulted in intracellular adenosine triphosphate levels of 96.98%, 88.44%, 43.3% and 27.56% relative to the control level respectively.

CONCLUSIONMCT1 can enhance the sensitivity of breast cancer cells to 3-BrPA possibly by transporting 3-BrPA into cells to inhibit cell glycolysis.