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.

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 investigate the characteristics of the cultured cervical cancer cell line transfected with anti HPV16E6 ribozyme, and to investigate the possibility and practicality of ribozyme in treatment of cervical cancer. Methods: The anti HPV16E6 ribozyme and empty eucaryotic expressing plasmids were transfected by lipofectin transfection into CaSKi cell, which named as CaSKi R, CaSKi P respectively. The morphology and the soft agar forming ability were studied. The expression of E6, PCNA and C erbB 2 genes was studied through Flow Cytometry. The tumorgenicity of each cell was detected by injecting cells into the nude mice skin. Three groups of nude mice were injected by CaSKi, CaSKi R and CaSKi P cell separately. Another group of mice was injected by CaSKi cell on right side and CaSKi R cell on left side. Results: There is no distinct difference of the morphology and growth rate between CaSKi and CaSKi P, but the growth rate of CaSKi R decreased. The soft agar forming rate of CaSKi P was similar with that of CaSKi cells, while that of CaSKi R was found decreased. The result of flow cytometric analysis showed that anti HPV16E6 ribozyme could reduce the expression of E6, PCNA and C erbB 2 genes on CaSKi R cells, while this phenomenon was not found on the CaSKi P cells. The tumorgenicity of CaSKi R in nude mice was decreased compared with CaSKi and CaSKi P cells. Conclusion: Anti HPVE6 rivozyme could partly reverse the malignant phenotypes of CaSKi cells. The reason may be the decrease of E6 gene expression, and the succeeding decrease of the PCNA and C erbB 2 genes′ expression.

This study was to observe the anti apoptosis effect of NADH on a normal liver cell line inflicted with chemical hypoxia. L02 liver cells were treated with 3mmol/L KCN in the presence of different concentrations of NADH (0～600?g/ml) for 0.5, 2, 4 hours. The percentage of apoptotic cells was assayed with flow cytometry(FCM). Another three groups of experiment were done as following. Group Ⅰ was control. Groups Ⅱ and Ⅲ were treated with 3mmol/L KCN in the presence or absence of NADH (400?g/ml) for 2h. The expression of proteins of Bcl XL, Bcl 2 and Bax in the L02 cells were determined with FCM analysis. The results showed that NADH could obviously prevent L02 cells from apoptotic damage at the concentration of 400?g/ml. it could also significantly upregulate the expression of Bcl 2 and Bcl XL proteins and downregulate the expression of Bax proteins after cyanide challenge( P

To investigate the role of NADH in preventing apoptotic damage of human hepatic cells induced by chemotherapy drug cisplatin, the ultrastructural changes in hepatic cells were examined with transmission and scanning electron microscopy, propidium iodide (PI) staining was used to measure the apoptotic rate with flow cytometry, the expressions of p53 and bcl 2 gene were detected with RT PCR, the change in caspase 3 and caspase 8 of apoptotic molecules was examined with colorimetric assay. The results showed that, compared with the group of cisplatin, morphological apoptotic changes were not obvious, the apoptosis rate was significantly decreased, p53 mRNA expression was decreased, bcl 2 mRNA expression was increased, caspase 3 and caspase 8 activity levels were kept in a low level in the group of NADH /DDP. The study indicates that NADH can prevent apoptosis of human hepatic cells, and increase the possibility of using NADH to reduce side effects of chemotherapy.

To study the repairing effect of NADH on PC12 cells damaged by chemical drugs, we determined the cytotoxicity of chemical drugs (cisplatin, doxorubicin and rotenone) on PC12 cells with cell proliferation test and MTT assay. At the same time, we analyzed the cyclin protein, GTPase binding protein and transglutaminase Ⅱ content with flow cytometry(FCM). NADH significantly inhibited the cytotoxicity of chemical drugs on PC12 cells. Cisplatin,doxorubicin and rotenone upregulated the expression of cyclin D1 and down regulated the expression of cyclin A and B1 in PC12 cells. On the contrary, NADH increased the expression of cyclin A and cyclin B1 and decreased the expression of cyclin D1. NADH could upregulate the expression of GTPase binding protein and transglutaminase Ⅱ, which were decreased by chemical drugs in PC12 cells. It is suggested that NADH could inhibit the cytotoxicity of chemical drugs to PC12 cells, which are associated with the regulation of cyclin proteins, GTPase binding proteins and transglutaminase Ⅱ.

To elucidate the mechanism of mitochondrial damage induced by rotenone and the possible biological function of NADH in repairing mitochondrial damage of PC12 cells, cytotoxicity test, immunocytofluorescence and flow cytometric analysis were used to investigate the changes of cell proliferation genes (c myc, c erbB 2), apoptosis inhibition genes bcl 2, p53 tumor suppressor protein, cell immediate early gene (c fos) and proliferating cell nuclear antigen (PCNA) in PC12 cells before and after exposure to rotenone. The results showed that rotenone could significantly inhibit the proliferation rate of PC12 cells and expression of c erbB 2, c myc, p53, and bcl 2 in PC12 cells, NADH could restore the proliferation activity of PC12 cell damaged by rotenone by gene regulation. It is suggested that rotenone could induce PC12 cells apoptosis not only by regulating mitochondria phosphorylation process, but also by down regulating the expression of oncogene proteins (C erbB 2, c myc), anti apoptotic gene protein (bcl 2), p53 tumor suppressor gene protein, and upregulating the expression of the immediate early gene c fos. Regulation of bcl 2, c myc, c erbB 2 and p53 might be involved in the repair of mitochondrial damage of PC12 cells.