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.

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.

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 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.

The influences of varied concentrations of AG on the 8H-TdR(3H-Thymidine ) incorporation inhibition rate and the release of PGI2 and TXA2 were studied in cultured human and rat lymphocytes . The results indicated that the AG increased the 3H-TdR incorporation inhibition rate on the concentrations of 1.5? 10-7mol/L and 3?10-7mol/L. Using the RIA method, authors find that AG on the concentrations of 4 ? 10-7mol/L, 4 ? 10-3mol/L and 4 ? 10-11mo/L can significantly decrease the cultured lymphocytes TXA2 release while has no clear influences on PGI2 release

To investigate a simple method visualizing neutral glycosphingolipid (N-GSLs) with iodine vapor, N-GSLs were isolated from the normal brain tissue of human by a modificated method of Hakomori, and then analyzed by HPTLC and visualized respectively by spraying aniline-diphenylamine-H 3PO 4 reagent and immersing into a jar full of iodine vapor. The plate should be kept 110℃ for 5 min until the coloration emerged by using the first method,and the N-GSLs were slowly made visible with a blue coloration. The N-GSLs were immediately made visible with a red-brown coloration by using the second method, and the color visible on the plates could emerge again after vaporized. Moreover, the second method was more sensitive than the first. The location of individual N-GSLs was the same by using the two methods. It suggested that a new method for visualizing N-GSLs has been established, it is simple, economic, rapid, innocuity and more sensitive, and the results are reproducible.

To investigate the role of tumor multidrug resistance (MDR) associated with CMH in the immunological evasion of MDR tumor cells, CMH was separated from neutral glycolipids of KBv 200by column chromatography, and the expression of B7 in DC was detected by flow cytometry. The results showed that the expression of B7 in DC was inhibited by CMH, it suggested the tumor MDR associated with glycolipids may inhibit human immune response by its effects on DC.

In order to evaluate the relationship between MDR of ovarian cancer and expression of neutral glycosphingolipids(N-GSLs) in ovarian cancer cell lines, the effects of TAM and VRP on the growth of COC1/DDP were assayed by MTT method. N-GSLs of the cells were isolated and purified with the modified Hakamoris method and analysed by HPTLC. The results showed that the expression of N-GSLs was different between parent cell lines and resistant cell sublines, the level of CMH was higher in COC1/DDP than in COC1. TAM and VRP could render multidrug-resistant cells sensitive to chemotherapy, while the level of CMH concomitantly was sharply decreased. It suggested that the expression of N-GSLs is associated with MDR of ovarian cancer, and CMH may be a kind of MDR related glycolipids in ovarian cancer.

In order to study the molecular structure of Glob-N, a glioma inhibitor, Glob-N was separated from the membrane of human type O red blood cells, cow brain and normal human brain tissue respectively. Their structures were identified by using time-of-flight mass spectrometry, 1H-NMR and 13C-NMR. The results showed that the structure of Glob-N from the membrane of human type O red blood cells was proposed as globotetraosylceramide, while Glob-N from cow brain and normal human brain was globotriaoceramide, and the number of carbon atoms in the ceramide of the Glob-N was 27, 33 and 26 respectively. It suggested that Glob-N is a kind of neutral glycosphingolipids, and its molecular structure is different in different tissues.