The relationship between the morphological development and the contents of glycogen, LDH, SDH in the beating as well as beat arrested cardiocytes cultured in vitro was studied cytochemically in neonatal rats. The results showed that. (1) After the cardiocytes were inoculated, they did not start to beat until they developed to elongated cells with some processes or stellated cells, and the contents of glycogen, LDH and SDH in cardiocytes reached to a given threshold (score) respectively. In beat arrested cells the contents of glycogen, LDH and SDH were below the beat-starting thresholds. (2) The greater the cell density of cardiocytes was, the more strongly the cell clusters beated. The cell density of cardiocytes was increased by mitosis. In 48-72h, cell divisions occurred in part of cardiocytes. Most mitotic figures were observed in 120h. When mitosis was in progress, cardiocytes containing glycogen particles became short but still with some branched processes. As the cardiocyte clusters were formed, the beat occurred strongly. The cytochemical reactions mentioned above may be used to evaluate how serious the cell is damaged in cytotoxicological and pharmalogical research.

The Epstein-Barr virus latent membrane protein 1 (LMP1) oncopro tein causes multiple cellular changes, including activation of the NF-κB trans cription factor. To elucidate its possible mechanism, the interaction between LM P1 and the tumor necrosis factor receptor associated factor (TRAF) molecules was detected by the immunoprecipitation-Western blotting assay. Results showed tha t LMP1 was co-precipitated with TRAF1,2,3 in the LMP1-HNE2 cell line. In the m eantime, κB reporter gene analysis revealed that over expression of TRAF1 or TR AF2 augmented LMP1-mediated NF-κB activation from LMP1, suprisingly, overexpr ession of either TRAF3 or an dominant negative TRAF3 inhibited the NF-κB activ ation, indicating that TRAF1 or TRAF2 is a positive modulator of LMP1-mediated NF-κB activation, whereas,TRAF3 is a negative modulator. Rather both CTAR1 (carboxy-terminal activating region 1) and CTAR2 domains of LMP1 can independently activate NF-κB by interacting with TRAF proteins. These data indicate that LMP1 interacts TRAF1,2,3 which are important for LMP1-mediated N F-κB activation, and further suggest that signaling from TRAFs may be involved in the progression to malignancy in cells of epithelial origin such as nasophar yngeal carcinoma (NPC).

Objective To study the characteristics of clustering of cardiovascular risk factors in patients less than 50 years-old of premature stable coronary heart disease(PSCHD)complicated with nonalcoholic fatty liver(NAFL).Methods One hundred and six patients with documented PSCHD were recruited into this study and their clinical data,including biochemical parameters,high-sensitivity C-reactive protein(hsCRP),white blood cell(WBC)count,ete.,were analyzed based on whether they had NAFL by B-type ultrasound scanning and their homeostasis model assessment ratio(Homa-IR)by the criteria for metabolic syndrome formulated by the International Diabetes Federation.Results Thirty-two (30.1percent)of 106 patients of PSCHD complicated with NAFL,and 74(69.9 percent)without NAFL. As compared to patients without NAFL,patients with NAFL had higher fasting blood glucose(FBS),serum level of insulin(INS),total cholesterol(TC),triglyceride(TG),serum activity of alanine aminotransferase(ALT),hsCRP,WBC count,body mass index(BMI),Homa-IR,and higher proportion of those with abnormal blood glucose,hypertension.metabolic syndrome(MS)and carotid atherosclerosis (CA)(P＜0.05),respectively.Bi-variate correlation analysis revealed that hsCRP positively correlated to BMI,TG,ALT and Homa IR(r=0.420,P=0.000;r=0.200,P=0.040;r=0.218,P=0.048:and r=0.546,P=0.000,respectively)and inversely correlated with serum level of high-density lipoprotein cholesterol(HDL-C)(r=-0.220,P=0.023).WBC count positively correlated with FBS(r=0.211,P=0.030).BMI,hsCRP,ALT,and proportions of hypertension,diabetes,MS,NAFL and CA in patients with Homa-IR above median were significantly higher than those in patients with that below median ( P＜0.05,respectively).Conclusions More risk faetors for chronic inflammatory reaction,cardiovascular disease and insulin resistance were clustered more obviously in patients of PSCHD complicated with NAFL.

Objective To investiagate cell apoptosis and expressions of Bax,Bcl-2 andCaspase-3 in gambogic acid-treated colorectal cancer cells. Methods SW480/LOVO colorectal cancer cells were treated by gambogic acid. Cell Counting Kit-8 assay (CCK-8) was used to test cell proliferation. Microscopy was used to check the morphological changes. Immunofluorescence staining technique was used to detect cell apoptosis. Expressions of Bax,Bcl-2 and Caspase-3 protein were detected by Western blot assay. Results Gambogic acid inhibited the proliferation of SW480/LOVO in a dose and time-dependent manner. Gambogic acid could induce cell apoptosis. Gambogic acid increased expressions of Caspase-3 and Bax, increased the ratio of Bax/Bcl-2, and decreased Bcl-2 protein expression. Conclusion Gambogic acid can inhibit proliferation and induce apoptosis of SW480LOVO cells, with the mechanism of up-regulation of Bax/Bcl-2 and activation of Caspase-3.

AIM: To examine the effects of recombinant human hepatocyte growth factor(rhHGF) and native calf HGF(cHGF) on SMMC-7721 human hepatocellular carcinoma(HCC) cell line. METHODS: Human HCC cell line culture, photometric assay, and flow cytometric assay were used in this study . RESULTS: A similar type of dose-dependent cell growth inhibition effect on SMMC-7721 human HCC cells by rhHGF(5-20 ?g/L) as well as by cHGF(25-100 mg/L) had been found, with the maximal effect at the highest concentration used. Approximately over 50% of the cells treated with rhHGF(5 ?g/L, 10 ?g/L, 20 ?g/L) accumulated in the quiescent G 0/G 1 phase of the cell cycle over incubation periods for 3 d. CONCLUSION: The growth of SMMC-7721 human HCC cells was strongly inhibited by both rhHGF and cHGF. This might be because the cells exposed to HGF became arrested in the G 0/G 1 phase.

OBJECTIVETo clarify if Epstein-Barr virus encoded LMP1 induces matrix metalloproteinase 9 expression via NF-kappa B or AP-1 signaling pathway, which gives evidence to the elucidation of the mechanism of LMP1- mediated carcinogenesis.

METHODSTo determine whether LMP1 or its mutants contribute to MMP9 production via NF-kappa B or AP-1 transcription factor, MMP9-chloramphenicol acetyl transferase (CAT), NF-kappa B mut 9-CAT, AP-1 mut MMP9-CAT were transfected into human nasopharyngeal carcinoma cells stably expressing LMP1 (HNE2-LMP1) or its mutants, [HNE2-LMP1 (1-185), HNE2-LMP1 (1-231), HNE2-LMP1 delta 187-351] by electroporation technic. The difference of MMP9 reporter activity among those cell lines was detected by CAT assay and expression of MMP9 was determined in nasopharyngeal carcinoma cells stably expressing LMP1 or its mutants by zymographic analysis. In the meantime, efforts were made to demonstrate if LMP1 regulates NF-kappa B or AP-1 activation using reporter gene analysis.

RESULTSIn contrast with vector-transfected cells, MMP9 CAT activity in HNE2-LMP1, HNE2-LMP1 (1-185), HNE2-LMP1(1-231), HNE2-LMP1 delta 187-351 increased 7.2, 1.3, 3.3, 4.0 times respectively. Zymographic analysis demonstrated that the 92 kDa MMP9 expression was induced in HNE2-LMP1, HNE2-LMP1(1-231) and HNE2-LMP1 delta 187-351 cells, whereas it was negative in HNE2-pSG5 and HNE2-LMP1 (1-185) cells. As compared to the HNE2 cells, NF-kappa B or AP-1 reporter activity in HNE2-LMP1 cells were increased 13.8, 8.4 fold respectively. Moreover, In contrast with MMP9 CAT-transfected cells, MMP9 CAT activity in NF-kappa B mut MMP9-CAT or AP-1 mut MMP9-CAT transfected HNE2-LMP1, HNE2-LMP1 (1-185), HNE2-LMP1(1-231) and HNE2-LMP1 delta 187-351 cells were significantly decreased by 18.1% or 16.3%, 35.0% or 33.3%, 29.1% or 26.1% from the original level. However, there was no difference in NF-kappa B mut MMP9-CAT or AP-1 mut MMP9-CAT transfected HNE2-pSG5, HNE2-LMP1 (1-185) cells.

CONCLUSIONIn nasophargyngeal carcinoma, Epstein-Barr virus-encoded LMP1 induces MMP9 transcription and enzymatic activity via an NF-kappa B or AP-1 signaling pathway, which may contribute to invasiveness and metastasis.

Gene Expression ; drug effects ; Herpesvirus 4, Human ; chemistry ; Humans ; Matrix Metalloproteinase 9 ; biosynthesis ; NF-kappa B ; metabolism ; Nasopharyngeal Neoplasms ; pathology ; Signal Transduction ; Transcription Factor AP-1 ; metabolism ; Tumor Cells, Cultured ; Viral Matrix Proteins ; pharmacology

OBJECTIVE: To investigate the association of myocardial blood flow (MBF) quantified by dynamic computed tomography (CT) myocardial perfusion imaging (MPI) with troponin level and left ventricle (LV) function in patients with ST-segment elevated myocardial infarction (STEMI). MATERIALS AND METHODS: Thirty-five STEMI patients who successfully had undergone reperfusion treatment within 1 week of their infarction were consecutively enrolled. All patients were referred for dynamic CT-MPI. Serial high-sensitivity troponin T (hs-TnT) levels and left ventricular ejection fraction (LVEF) measured by echocardiography were recorded. Twenty-six patients with 427 segments were included for analysis. Various quantitative parameters derived from dynamic CT-MPI were analyzed to determine if there was a correlation between hs-TnT levels and LVEF on admission and again at the 6-month mark. RESULTS: The mean radiation dose for dynamic CT-MPI was 3.2 ± 1.1 mSv. Infarcted territories had significantly lower MBF (30.5 ± 7.4 mL/min/100 mL versus 73.4 ± 8.1 mL/min/100 mL, p < 0.001) and myocardial blood volume (MBV) (2.8 ± 0.9 mL/100 mL versus 4.2 ± 1.1 mL/100 mL, p = 0.044) compared with those of reference territories. MBF showed the best correlation with the level of peak hs-TnT (r = −0.682, p < 0.001), and MBV showed a moderate correlation with the level of peak hs-TnT (r = −0.437, p = 0.026); however, the other parameters did not show any significant correlation with hs-TnT levels. As for the association with LV function, only MBF was significantly correlated with LVEF at the time of admission (r = 0.469, p = 0.016) and at 6 months (r = 0.585, p = 0.001). CONCLUSION: MBF quantified by dynamic CT-MPI is significantly inversely correlated with the level of peak hs-TnT. In addition, patients with lower MBF tended to have impaired LV function at the time of their admission and at 6 months.
Blood Volume ; Echocardiography ; Heart Ventricles ; Humans ; Infarction ; Myocardial Infarction ; Myocardial Perfusion Imaging ; Reperfusion ; Stroke Volume ; Troponin T ; Troponin

Advanced glycation end products (AGEs) have been implicated in aging and aging related diseases. Therefore, the clinical detection of AGEs is of great importance for the early prediction, intervention and long-term monitoring of above mentioned chronic diseases. There are various detection methods for assessment of AGEs, but due to the great heterogeneity and complex structure of AGEs, there is a lack of standardized detection method for AGEs so far. This review summarizes the current detection methods for AGEs and their advantages and disadvantages, aiming to highlight the future directions for the clinical detection of AGEs.

OBJECTIVETo investigate the association of gankyrin protein expression in colorectal cancer (CRC) with its prognosis.

METHODSClinical data and resection samples of 100 colorectal cancer patients identified by pathology undergoing resection in our department from June 2008 to June 2009 were collected. The gankyrin expression in CRC tissues and matched adjacent noncancerous tissues collected during the operation of 100 CRC cases was detected by immunohistochemical staining and Western blotting. The associations of gankyrin expression level with overall survival, clinicopathologic features were analyzed by Chi square test, Cox regression analysis, Kaplan-Meier analysis and log rank test.

RESULTSImmunohistochemical staining showed that the positive brown granules were mainly distributed in the cytoplasm, and nuclear immunostaining was observed in tissue samples of 29 cases, of whom 16 cases had distal metastasis [55.2% (16/29)]. The positive rate of gankyrin and the relative gray value of Western blotting in CRC tissues were 67% (67/100) and 0.69±0.23, respectively, which were significantly higher than those of 2 cm adjacent noncancerous tissues [6% (6/100) and 0.31±0.16] and 10 cm adjacent noncancerous tissues [1%(1/100) and 0.16±0.11] (all P<0.001). Patients with positive expression of gankyrin had worse survival than those with negative ones (41.8% vs. 72.7%, P=0.008). The gankyrin expression was associated to lymph node metastasis (P=0.005), tumor stage (P=0.001) and distal metastasis (P=0.002). Cox regression analysis showed that distal metastasis (P=0.004) and high expression of gankyrin (P=0.038) were independent risk factors for poor prognosis of patients with CRC.

CONCLUSIONUp-regulated expression of gankyrin is related to invasion and metastasis of human CRC, and gankyrin may be valuable in predicting prognosis.

Blotting, Western ; Colorectal Neoplasms ; Humans ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Postoperative Period ; Prognosis ; Proteasome Endopeptidase Complex ; Proto-Oncogene Proteins

OBJECTIVETo elucidate the regulation of the phosphorylation of epidermal growth factor receptor (EGFR) by the EB virus encoded latent membrane protein 1 (LMP1) in nasopharyngeal carcinoma cell line.

METHODSThe levels of EGFR expression and phosphorylation in pTet-on LMP1 HNE2 cell, a nasopharyngeal carcinoma (NPC) cell line, in the dynamic expression of LMP1 induced by different concentrations of doxycycline (Dox) were observed. The EGFR dominant negative mutant and LMP1 antisense expression plasmid were transiently transfected into pTet-on LMP1 HNE2 cells by lipofectamine, and the changes in EGFR phosphorylation were observed by immunocoprecitation and Western blot. The changes in EGFR phosphorylation were observed after EGF treatment.

RESULTSIn pTet-on LMP1 HNE2 cells, Dox-induced LMP1 upregulated EGFR expression and phosphorylation in a dose-dependent manner. After EGFR dominant negative mutant was transfected into pTet-on LMP1 HNE2 cells, the increase of EGFR phosphorylation was inhibited completely. When LMP1 antisense expression plasmid was transfected into pTet-on LMP1 HNE2 cells, the levels of EGFR phosphorylation were also inhibited significantly. Meanwhile, after EGF had been added into pTet-on LMP1 HNE2 cells, increase of EGFR phosphorylation was induced, but it was completely blocked by EGFR dominant negative mutant and the introduction of LMP1 antisense.

CONCLUSIONEB virus encoded LMP1 not only induces the dose-dependent expression of EGFR, but also the dose-dependent phosphorylation of EGFR. The phosporylation of EGFR may play a vital role in the development of nasopharyngeal carcinoma.

Blotting, Western ; Epidermal Growth Factor ; metabolism ; Herpesvirus 4, Human ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; pathology ; virology ; Phosphorylation ; Receptor, Epidermal Growth Factor ; metabolism ; Tumor Cells, Cultured ; Viral Matrix Proteins ; metabolism