Objective To investigate molecular diagnostic method for hereditarymethemoblobinemia. Methods The cDNA coding sequence of NADH-cytochrome b5 reductase (b5R) from 3 patients with hereditary methemoglobinemia was analyzed by direct sequencing of RT-PCR products and the genomic DNA of b5R gene by PCR-restriction endonuclease digestion or PCR-sequencing. Results The b5R cDNA of patient A was T/C heterozygous at nucleotide 527 and G/A heterozygous at nucleotide 608. The b5R cDNA of patient B was G/A heterozygous at both nucleotide 170 and nucleotide 179. The b5R cDNA of patient C was G/A heterozygous at nucleotide 608 and C/T heterozygous at nucleotide 791. Result of genomic DNA analysis was in agreement with that of cDNA approach. Conclusion The method for molecular diagnosis of hereditary methemoglobinemia was established and 3 novel b5R gene mutations were identified in compound heterozygosity in 3 Chinese patients.

For obtaining new structural compounds with unique resistance profiles or novel mechanisms of action on HIV-1 from natural products, anti-HIV-1 drug screening models were used in vitro. Norcantharidin (NCTD), a derivative from cantharidin, was found to have inhibitory activities on HIV-1(IIIB) p24 antigen in lymphocyte lines MT-4, CEM and H9. It inhibited HIV-1 strain 018a (sensitive to zidovudine) from replicating with EC50 (50% effective concentration) of 14.9 micromol L(-1) and also inhibited HIV-1 strain 018c (resistant to zidovudine) from replicating with EC50 of 20.2 micromol L(-1) in primary lymphocytes peripheral blood mononuclear cells (PBMC). Norcantharidin showed synergistic activity with zidovudine on HIV-1(IIIB) in MT-4 cells, the combination index was less than 0.3. But, it was not active on HIV-1 integrase, reverse transcriptase or protease in vitro. As the structure of norcantharidin is unique and different from that of all clinic drugs approved, it would be possible to obtain new and effective compounds against HIV-1 with low toxicities after modification of norcantharidin.

Objective To approach the proliferative activities of Hodgkin's disease cells were transfected with mtrⅡ gene.Methods The ki67 and PCNA expression of Hodgkin's disease cells and those which were transfected with mtrⅡ gene by immunohistochemical technique respectively.Results The expression rate of ki67 and PCNA in Hodgkin's disease cells which were transfected with mtrⅡ gene was higher than that weren't transfected(P

Objective To investigate the proliferative activities and significance of Hodgkin's disease cells which were transfected with mtr Ⅱ gene. Methods The ki67 and proliferating cell nuclear antigen (PCNA) expression in Hodgkin's disease cells transfected with mtr Ⅱ gene were determined with immunohistochemistry. Results The ki67 and PCNA expression in cells transfected with mtr Ⅱ gene was more than that of cells without mtrⅡ gene transfected (P<0.05). Conclusion The proliferative activities of Hodgkin's disease cells transfected with mtr Ⅱ gene increased obviously, which may be related to the progress of Hodgkin's disease.

The effectiveness of traditional Chinese medicine (TCM) against various diseases urges more low cost,speed and sensitive analytical methods for investigating the phamacology of TCM and providing a theoretical basis for clinical use.The potential of second-order calibration method was validated for the quantification of two effective ingredients of Schisandra chinensis in human plasma using spectrofluorimetry.The results obtained in the present study demonstrate the advantages of this strategy for multi-target determination in complex matrices.Although the spectra of the analytes are similar and a large number of interferences also exist,second-order calibration method could predict the accurate concentrations together with reasonable resolution of spectral profiles for analytes of interest owing to its ‘second-order advantage'.Moreover,the method presented in this work allows one to simply experimental procedure as well as reduces the use of harmful chemical solvents.

Objective To study the detection of Na+/H+exchanger regulatory factor 3(NHERF3) protein expression in renal carcinoma and its correlation with malignant biological behavior. Methods Renal clear cell carcinoma and their adjacent tissues of forty- eight patients were collected. Immunohistochemical method was used to detect the positive expression of NHERF3 protein,and specific expression was detected by Western-blot.Patients were further divided into high NHERF3 group and low NHERF3 group according to median expression of NHERF3 protein,and each group had 24 cases.The expressions of proliferation,invasion and autophagy genes in tumor tissues were detected by fluorescent quantitative PCR.Results The positive rate of NHERF3 protein and the expression of NHERF3 protein in renal carcinoma tissue was significantly lower than that in paracancerous tissue(P<0.05).Expressions of proliferation genes such as k-Ras,c-Myc,TRPC1 mRNA in low NHERF3 group were higher than those in high NHERF3 group:141.74 ± 18.95 vs. 100.00 ± 0.00, 135.88 ± 16.32 vs. 100.00 ± 0.00, 137.21 ± 16.98 vs.100.00 ± 0.00;MIIP,FOXO1 mRNA levels were lower than those in high NHERF3 group: 43.19 ± 5.88 vs. 100.00 ± 0.00, 38.76 ± 4.51 vs. 100.00 ± 0.00; expressions of invasion genes such as CD74, Fascin, MACC1, TRPM8 mRNA were significantly higher than those in high NHERF3 group:152.18 ± 17.64 vs. 100.00 ± 0.00, 146.29 ± 17.63 vs. 100.00 ± 0.00, 139.76 ± 15.82 vs. 100.00 ± 0.00,150.47 ± 17.95 vs.100.00 ± 0.00;expressions of autophagy genes such as Beclin-1,LC3 mRNA were significantly lower than those in high NHERF3 group: 63.21 ± 7.09 vs. 100.00 ± 0.00, 56.28 ± 7.15 vs. 100.00 ± 0.00; EZH2 mRNA level was higher than that in high NHERF3 group:159.47 ± 17.82 vs.100.00 ± 0.00,and there were significant differences(P<0.05).Conclusions The positive rate of NHERF3 protein expression and the amount of protein expression in renal carcinoma tissue is increased, and the specific expression is closely related to tumor proliferation, invasion and activity of autophagy.