To investigate the roles of neutral glycosphingolipids (N-GSLs) in the development of tumor, tumor antigen expression and immunological evasion, the regularities of the expression of N-GSLs in human embryonic and neoplastic tissue were studied, the influence of N-GSLs in the growth regulation, immunological evasion and multidrug resistance(MDR) of tumor cells were analyzed. The tumor embryonic associated antigen CDH, tumor multidrug resistance associated N-GSLs CMH and neoplasm inhi-bitor globoside were found. The significance of N-GSLs synthesis inhibition and de-N-glycosylation in the reversion of MDR and tumor therapy was also partly disclosed.

The author investigated the function of coenzyme NADH in increasing the level of energy metabolism, repairing cellular damage, improving cellular stress ability, decreasing cytotoxicity of chemotherapy drugs and radiation against normal tissue. The molecular regulation mechanism of NADH in cytoprotection was elucidated. A new prevention and cure way was provided in cytoprotection treatment for clinical disease.

Objective: To investigate the diagnostic value of combined examination of copeptin and high sensitivity cardiac troponin T (hs-cTnT) in patients at the early stage of acute myocardial infarction (AMI) . Methods: A total of 272 patients were enrolled in this study, all of them suffered from chest pain and admitted within 4 hours. The patients were divided into 4 groups according to coronary artery angiography (CAG) results. Control group, the patients with normal CAG,n=64, UAP group (unstable angina pectoris),n=50, STEMI group,n=82, NSTEMI group,n=76. All patients received in-hospital observation, plasma levels of copeptin and hs-cTnT were examined at admission and at 6 hours after the chest pain respectively. Results: Within 4 hours of chest pain, combined examination of copeptin and hs-cTnT had the higher sensitivity for diagnosing AMI than a single detection of hs-cTnT with the cut-off point of hs-cTnT ≤ 14ng/L and Copeptin < 14pmol/L. In NSTEMI group, the AUC (area under curve) for combined examination was 0.97 (95% CI 0.88-0.99), AUC for single hs-cTnT detection was 0.75 (95% CI 0.62-0.87),P<0.05. In STEMI group, the AUC for combined examination was 0.97 (95% CI 0.88-0.99), AUC for single hs-cTnT detection was 0.74 (95% CI 0.60-0.88),P< 0.05. The AUC for combined examination of copeptin and hs-cTnT in diagnosing early AMI was 0.912 (95% CI 0.812-0.961) which was higher than single detection of hs-cTnT, AUC 0.851 (95% CI 0.713-0.936), Z=2.553,P<0.05. Conclusion: Combined examination of copeptin and hs-cTnT had the higher sensitivity and accuracy for diagnosing the patients at the early stage of AMI, it may help the risk stratiifcation of chest pain which is valuable in clinical practice.

Objective: To study the characterization of the cultured cervical cancer cell line transfected with anti- HPV16E6-ribozyme, and to investigate the effect of ribozyme on proliferation and apoptosis of cervical cancer cell. Metliods: Anti-HPV16E6-ribozyme had been designed to cleave the HPV16E6 gene. With the method of lipofectin transfec- tion, the anti-HPVI6E6-ribozyme and empty eucaryotic expressing plasmids were transfected into CaSKi cell, which named as CaSKi-R, CaSKi-P respectively. The amounts of E6 mRNA in the three kinds of cells were detected by northern blot. Cell cycle was detemined by flow cytometry, and cell apoptosis was examined by fluorescent (Hoechst) staining and TUNEL. The expression of some genes, including c-myc, bcl-2, p53, and fas, was also detected by flow cytometry analy- sis. Results: Northern blot showed that E6 mRNA was less in CaSKi-R than in CaSKi. In CaSKi-R cells, cycle was arres- ted in G1 phase, with decreasing in percentage of S phase cells. The apoptosis rate of CaSK1-R cell was much higher than those of CaSKi and CaSK1-P. Anti-HPV16E6-ribozyme could reduce the expression of E6, c-myc, bcl-2 genes on CaSKi- R cells, and increased the expression of p53. While this phenomenon was not found on the CaSK1-P cells. The expression of fas was similar in the three kinds of cells. Conclusion: Anti-HPVE6-rivozyme induces apoptosis of human cervical cancer CaSKi cells. The mechanisms may be the decrease of E6 gene's expression, and the succedent changing of some genes'expression.

R 5 was administered in vitro to observe its antitumor activity in human breast carcinoma cell line MCF 7. As the most useful and efficient anthracycline, epirubicin(EPI) was served as the positive chemical treatment control. MTT colorimetric assay was applied to detect cytotoxicity of R 5 to MCF 7 cells. Apoptotic rate of cells double marked with Annexin V FITC and PI was examined by flow cytometry. And, the alteration of wild type (WT) p53, bax and bcl 2 proteins expression was also observed. Ultrastructural change of MCF 7 cells was observed under a transmission electron microscope. The results showed that growth restrain was observed in MCF 7 cells when administered with different doses of R 5 or EPI. The effect was increased concomitantly with the increasing of R 5 or EPI concentration and culture time. Apoptosis was observed since 6 hours after the MCF 7 cells cultured with R 5 or EPI, and the effect was increased with the culture time extending and reached the highest peak at about 48 hours. However, the apoptotic rate decreased when cultured for 72 hours. Different doses of R 5 and EPI can all induce apoptosis in MCF 7 cell, and the apoptotic rate increased with their concentration, but decreased when the concentration was higher than 5?10 -6 mol/L. Ultrastructure of apoptotic MCF 7 cells, observed by transmission electron microscope, showed typical morphologic changes of apoptosis.

Objective To study the modulation of mdr1 and P glycoprotein (P gp) by 1 phenyl 2 palmitoylamino 3 morpholino 1 propanol (PPMP) in SKOV3 adriamycin resistant (SKOV3/AdrR) cell line Methods SKOV3/AdrR cells were treated with PPMP, mRNA expression of multidrug resistant (mdr1) gene was analyzed by reverse transcriptase polymerase chain reaction Intracellular rhodamine (Rh123) concentration was measured by flow cytometry Results PPMP was found to inhibit mdr1 expression of SKOV3/AdrR at the mRNA level This modulation of gene expression was content dependent and complete inhibition appeared at 25 ?mol/L PPMP treatment PPMP could increase intracellular Rh123 accumulation in SKOV3/AdrR cells After 15, 25 ?mol/L PPMP treatment, Rh123 accumulation in SKOV3/AdrR was markedly enhanced Rh123 fluorescence intensity were 389 98,426 08 respectively ( P

Objective: To investigate the effects of HPV16E6-ribozyme on telomerase activity in cervical carcinoma cell line CaSKi and the related mechanisms. Methods: Anti-HPV16E6-ribozyme and blank eucaryotic plasmids were transfected into CaSKi cells via lipofectin, and the resultant cells were named as CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blotting. The expression of E6 mRNA and protein in the 3 kinds of cells were detected by Northern blotting and Western blotting, respectively. Telomerase activity was determined by TRAP-Elisa method; the expression of P53, c-myc, hTERT and hRT mRNA were examined by RT-PCR.Results: RNA dot blotting showed that anti-HPV16E6-ribozyme was stably expressed in transfected CaSKi-R cells. Western blotting showed that the expression of E6 mRNA and protein in CaSKi-R cells was obviously lower than that in CaSKi and CaSKi-P cells. The telomerase activities in CaSKi,CaSKi-P and CaSKi-R cells were (0.89?0.14), (0.90?0.11) and(0.36?0.06),respectively. The inhibitory rate of telomerase activity in CaSKi-R cells was 59.55%, which was significantly lower than those in CaSKi and CaSKi-P cells (P

The current study was designed to investigate the effects of PPMP (DL-threo-1-phenyl-2- palmitoylamino-3-morpholino-1-propanol), a kind of glycolipids synthase inhibitor, on the modulation of mdr1 mRNA expression and the reversing effect of multi-drug resistance by PPMP in human malignancy KBv200cell line. In vitro KBv200 cells were treated with PPMP in different concentration, the alterations of mRNA expression of drug-resistant gene mdr1 in KB (sensitive cell line) and KBv200 (before and after the treatment of PPMP) cells were analyzed by RT-PCR. Intracellular rhodamine(Rh123) concentration was measured by flow cytometry. PPMP was found to inhibit mdr1 gene expression of KBv200 at the mRNA level, and complete inhibition appeared at 25μmol/L PPMP treatment for 48h. PPMP could increase intracellular Rh123 accumulation in resistant cell lines. This modulation of gene expression and Rh123 accumulation was directly correlated with the concentration of PPMP. It suggested that PPMP, a chemical inhibitor of glycolipids synthase, could modulate mdr1 expression at the mRNA level in a content dependent manner, PPMP possesses MDR-reversing activity.

To probe the effect of reduced form coenzyme Ⅰ(NADH) in antagonizing cardiac muscle toxicity induced by doxorubicin and its underlying mechanism. Primary culture of myocardial cells of SD rat and doxorubicin injury model were established. MTT assay, laser confocal microscopy, transmission electron microscopy and biological oxygen monitor were used to observe the morphology and function of mitochondria. The results showed that the killing rate was increased in the group of doxorubicin, and that in the group of NADH/doxorubicin was greatly decreased. Doxorubicin could induce ultrastructural damage of cardiomyocyte mitochondria, manifested as swelling, disintegration, disruption of cristae and fusion. Cardiac mitochondria were protected against injuries in the group treated with NADH/doxorubicin. There was a significant difference in the fluorescence intensity of mitochondria membrane potentional and ROS between the groups treated with doxorubicin and NADH/ doxorubicin. It is suggested that NADH can significantly antagonizing cardiac muscle toxicity induced by doxorubicin and can protect mitochondrial structure and function.

AIM To study effects of antioxidant NADH on damage of irradiated normal cell lines. METHODS L02 liver cells were cultured in RPMI 1640, exposed to X ray irradiation, and continued to culture in the presence or absence of NADH for 24 h. The cellular viability was determined by routine MTT method. Using fluorescence probe and confocal microscope, the level of cellular H 2O 2 was detected. Positive rate of bcl 2 and Bax protein expression in the L02 cells were analyzed by flow cytometry. RESULT NADH can not only antagonized growth inhibition of X ray irradiated L02 cells, decreased cellular H 2O 2 production, but also significantly increased positive rate of L02 cells expressed bcl 2 protein and decrease positive rate of L02 cells expressed Bax proteins ( P