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 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.

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.

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.

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.

Purpose:To observe the effect and safety of c om bination therapy with Percutaneous Local Cryotherapy(PLCT)and Percutaneous Etha nol Injection Therapy( PEIT)in patients with irresectable hepatic cancer Methods:43 patients with irresectable hepatic carcinoma were enrolled 29 tumors of 14 patients were treated by PLCT,23 tumors of 16 patients were treated by PEIT,and 23 tumors of 13 patients were treated by combination therapy Results :The tumor size decreased by more than 50% 1 month later in 51 7%(15/29 )of tumors treated by PLCT alone, in 43 5%(10/23)of tumors treated by PEIT alone, 78 3%(18/23)tumors treated by combination therapy The 1-year surv ival rates were 64 3%(9/14) in PLCT group?43 8%(7/16) in PEIT group?8 4 6%(11/13)in combination therapy group The blood levels of AFP decreased by various extents Conclusions:Combination therapy with PLCT a nd PEIT is more effective than PLCT or PEIT alone

BACKGROUND: Tissue-engineered artificial nerve was successfully constructed with the compound of acellular nerve graft and bone marrow mesenchymal stem cells, suggesting that it could promote peripheral neural regeneration.OBJECTIVE: To construct tissue-engineered artificial nerve, and to verify neural functional recovery of bridging rats following sciatic nerve defect.METHODS: A total of 60 adult male SD rats were used to induce sciatic nerve defect models (15 mm in length), and they were then randomly divided into three groups, with 20 rats in each group. Sciatic nerve defect group was treated with tissue-engineered artificial nerve; blank control group was treated with tissue-engineered nerve stent; autoallergic neural control group was treated with autoallergic neural transplantation. Twelve weeks after bridging, histology of sciatic nerve and neuralfunctional recovery were detected via gross observation, wet mass of tibialis anterior muscle, and histological analysis.RESULTS AND CONCLUSION: At 12 weeks after bridging surgery, rats in experimental group were able to stand on the floor,and withdrawal reflex was detected at plantar skin on the surgical side. S-100 protein of plantar skin was positive. There was no significant difference in wet mass of tibialis anterior muscle between experimental and autoallergic neural transplantation group (P ＞ 0.05). HRP retrograde tracing in the experimental group demonstrated that HRP-positive cells were observed in both spinalcord and posterior root ganglion. There was no significant difference in number of myetinated nerve fiber, thickness of myelin sheath, and area of nerve tissue between experimental and autoallergic neural transplantation group. The results demonstrated that the compound of acellular nerve graft and bone marrow mesenchymal stem cells could successfully construct tissue-engineered artificial nerve to repair sciatic nerve defect and promote neurohistological reconstruction and functional recovery.

Objective To establish a stable cell line expressing transmembrane form of human blood group A antigen mimotope vaccine by transfecting malignant melanoma cell line B16, and to detect the cytotoxicity of the vaccine against melanoma cells. Methods Cultured B16 cells were classified into 4 groups, i.e.,P/F-M-pIRES group [transfected with the recombinant plasmid mimotope peptide/Fas-macrophage inflammatory protein (Mip)-pIRES], P/F-pIRES group (transfected with the recombinant plasmid mimotope peptide/FaspIRES), M-pIRES group (transfected with the recombinant plasmid Mip-pIRES), and pIRES group (transfected with the empty plasmid pIRES). B 16 cells were transfected through Lipofectamine 2000. Subsequently, RT-PCR and Western blotting were performed to detect the mRNA and protein expressions of the mimotope peptide/Fas fusion gene and Mip3β in transfected B16 cells. Cell counting kit-8 (CCK-8) was used to evaluate the vaccinemediated complement dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC)against B16 cells. Results RT-PCR yielded specific DNA fragments with expected size. Western blotting revealed the anti-A antibody-binding activity of the recombinant mimotope peptide/Fas fusion protein. Factor analysis indicated significant differences in CDC (F = 244.522, P ＜ 0.01 ) and ADCC (F = 71.593, P ＜ 0.01 )against B16 cells between the 4 groups. Group comparisons demonstrated more intense CDC and ADCC in P/FM-pIRES and P/F-pIRES groups compared with M-plRES and pIRES groups, stronger ADCC in P/F-M-pIRES group in comparison with P/F-pIRES group (F = 15.42, P ＜ 0.05), but no significant difference in CDC was observed between M-pIRES and pIRES group. Conclusions The transmembrane form of human blood group A antigen mimotope vaccine could be stably expressed in B16 cells, and mediate ADCC and CDC against B16 cells in vitro.