Objective To investigate the role of NF-kB in cytokines (IL-1?, TNF-? and IFN-r) -induced impairment of INS-1 cell, a pancreatic islet ?-cell line. Methods Retrovirus expressing IkB??N with mutant IkB?(inhibitor of NF-kB ) was constructed. Immunoblot analysis, fluorescence analysis using a kB-1uc reporter gene and thiazolyl blue viability assay were applied in this study. Results IL-1?-induced IkB? degradation and NF-kB activation were found in INS-1 cells but not in INS-1/IkB??N cells infected with IkB??N retrovirus. The viability of INS-1 cells incubated with the combination of IL-1? and IFN-r or TNF-? and IFN-r was decreased but INS-1/IkB??N cells could resist these cytokines-induced decrease of cell viability. Conclusion IL-1?, TNF-?can decrease the viability of? cells, while IFN-r may have some enhancing role. Inhibition of NF-kB activation can protect ? cells from the cytotoxicity of the cytokines.

Opjective To study the prevalence of diabetic nephropathy of type 2 diabetes and its correlated risk factors. Methods 1 059 cases with type 2 diabetes in our university hospital during the period of 1997-2000 were screened for diabetic nephropathy,and its correlated risk factors were analysed. Results The prevalence of diabetic nephropathy and end stage of renal disease were 9.73% and 5.47%, respectively. Hypertension and dyslipidemia characterized with hypertriglycemia and low leve1 of HDL were the main risk factors of type 2 diabetic nephropathy. There was no significant relationship between single-determining HbAlc level and UAER. Conclusion The prevalence of diabetic nephropathy of this study was slightly lower than that reported by other authors in western countries,and the risk factors were consistent with those papers.

Objective To investigate the effects of host factors and hepatitis B virus (HBV) factors on interferon treatment in patients with chronic hepatitis B(CHB). Methods Polymerase chain reaction sequence specific primers (PCR SSP) technology was used to determine human leukocyte antigen (HLA) DR and DQ alleles in 32 patients with CHB. Serum HBV DNA level was quantified by a PCR assay with a low limit of detection of 1000 copies/ml. HBV genotypes, precore(A 1896 ) and core promoter(T 1762 A 1764 ) mutations was determined by PCR restriction fragment length polymorphism(PCR RFLP) analysis. Results The frequency of HLA DRB1*14 allele was higher in 10 responders to IFN treatment than in 22 non responders(20.0% v.s. 2.3%, relative risk(RR)=10.750, P =0.030); whereas that of HLA DQB1*07 is lower(10.0% v.s. 38.6%, RR=0.176, P =0.022). Pretreatment serum HBV DNA level in 10 responders was lower than that in 22 non responders (6.71?1.06 v.s. 7.59?0.56log 10 copies/ml, P =0.030). Patients with HBV genotype B infection had a higher response rate to IFN than those with HBV genotype C infection(53.8% v.s. 15.8%, P =0.049). Pretreatment serum ALT level, presences of A 1896 mutation and T 1762 A 1764 mutations had no difference between responders and non responders. Logistic multivariate analysis identified low pretreatment HBV DNA level(RR=0.212, P =0.034), presence of A 1896 mutation(RR=0.063, P =0.049) and HLA DQB1*07(RR=13.358, P =0.045) as independent factors associated with IFN antiviral response. Conclusions Low load of pretreatment serum HBV DNA and T 1762 A 1764 mutation in HBV genome or HBV of genotype B are associated with the response rate to IFN treatment in patients with CHB.

Objective To explore the effect of the treating left colon cancer,Compare left colon irrigation at one-stage radical colon resections were performed with the way which is given colon bracket insertion operation then two-stage radical colon resections were performed.Methods 32 cases sigmoid and rectum cancer patients with obstruction were received.And 14 cases were treated by colon irrigation at one-stage radical colon resections; 18 cases were treated by colon bracket insertion operation then two-stage radical colon resections were performed.To analyze the complications of after-operation、the average time in hospital.Results No occurence of anastomotic leakage was found in the two-stage radical colon resections.There was only one case was infected; the operation time(115 ± 26)min.The drainage in 3 days after the operation(127 ± 66)ml,length time(12.5 ± 3.2)d.The group of one-stage radical colon resections:the operation time(210 ± 39)min.The drainage in 3 days after the operation.length time(14.8 ±2.6)d,There was two cases of anastomotic leakage and 12 cases were infected and two dead were found in this group.Conclusion The way which was given colon bracket insertion operation then two-stage radical colon resections were performed,was better than colon irrigation at one-stage radical colon resections were performed,and with the first way patients life quality enhanced.

Objective Trichophyton rubrum strains can cause superficial infection and also deep tissue infection, but relevant animal model has not been reported yet.The aim of this study was to construct an animal model of granuloma infected by T.rubrum. Methods Three T.rubrum strains isolated from clinical granuloma tissues, 2 T.rubrum strains isolated from tinea infection and a standard strain of ATCCMYA4438 were selected.Corticosteroids were given to the Balb/C mice before and after the injection of the T. rubrum and mucin suspension and the mice model of granuloma was established.Direct microcopy, culture and histopathologic method were adopt to verify the infection effects. Results The mice inoculated with the T.rubrum granuloma strains with mucin suspension were examined after 21 days in the condition of applying appropriate dose of glucocorticoids.Direct microscopic examination showed the slender mycelium, fungal culture showed the growth of colony and histopathology showed excessive keratinization of foot tissue, formation of granuloma in the dermis with inflammatory cell infiltration of neutro-philic granulocyte and lymphocytes.However, the mice inoculated with the T.Rubrum tinea strains with mucin suspension showed the negative result. Conclusion The rubrum granuloma mice model can be es-tablished using the clinical isolates of T.rubrum granuloma strains with the mucin and glucocorticoids interventions.

This study is to investigate the effect of high altitude hypoxia on the activity and protein expression of CYP2C9 and CYP2C19. Rats from plain (P) and rats with acute middle altitude hypoxia (AMH), chronic middle altitude hypoxia (CMH), acute high altitude hypoxia (AHH) and chronic high altitude hypoxia (CHH) were administered orally phenytoin sodium (PHT) and omeprazole (OMZ) to evaluate the activity of CYP2C9 and CYP2C19, separately. The serum concentrations of PHT and metabolite 4'-hydroxyphenytoin (HPPH) at 12 h after treatment and the serum concentrations of OMZ and metabolite 5-hydroxy omeprazole (5-OHOMZ) at 3 h after treatment were determined by RP-HPLC. The activity of CYP2C9 and CYP2C19 was evaluated by the ratio of HPPH to PHT and the ratio of 5-OHOMZ to OMZ, respectively. The protein expressions of CYP2C9 and CYP2C19 were determined by ELISA method. The activities of CYP2C9 (HPPH/PHT) in P, AMH, CMH, AHH and CHH were 0.67 +/- 0.31, 0.75 +/- 0.29, 0.76 +/- 0.23, 0.79 +/- 0.31 and 0.75 +/- 0.18, respectively, and the activities of CYP2C19 (5-OHOMZ/OMZ) in P, AMH, CMH, AHH and CHH were 0.17 +/- 0.06, 0.20 +/- 0.10, 0.11 +/- 0.05, 0.37 +/- 0.13 and 0.19 +/- 0.05, respectively. The protein expressions of CYP2C9 in P, AMH, CMH, AHH and CHH were 4.20 +/- 1.27, 3.95 +/- 0.81, 3.93 +/- 1.11, 4.32 +/- 1.03 and 4.12 +/- 0.86 ng x g(-1), respectively, and the protein expressions of CYP2C19 in P, AMH, CMH, AHH and CHH were 3.91 +/- 1.82, 3.63 +/- 2.07, 2.55 +/- 0.85, 4.78 +/- 2.37 and 3.51 +/- 1.03 ng x g(-1), respectively. The activities and protein expressions of CYP2C9 in AMH, CMH, AHH and CHH were not significantly different with those of P. The protein expressions of CYP2C19 in AMH, CMH, AHH and CHH were not significantly different with those of P, but the activity of CYP2C19 in AHH was significantly higher than that of P. This study found significant changes in the activity of CYP2C19 under the special environment of acute high altitude hypoxia.

Objective To investigate the possibility and feasibility of the whole genome microarray scanning technique in clinical cytogenetic diagnosis of an uncertain karyotype and mentally retarded child. Methods The karyotype analysis of the mental development delayed child was 47, XY+mar. Genomic DNA was extracted from the peripheral blood and the whole genome microarray scanning technique was used to analyze the derivative chromosome. Results The whole genome microar-ray scanning technique indicated the derivative chromosome fragment had originated from 9p13.1-p24.3. Conclusions Com-paring to conventional cytogenetic analysis methods, the whole genome microarray scanning technique is of high resolution, high-throughput and high accuracy, which can detect the submicroscopic chromosomal aberrations and replace the conven-tional karyotype analysis.

10.3969/j.issn.1000-3606.2013.06.019

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.

Objectives To develop a rapid genotyping method of clinical isolates of Malassezia from patients with tinea versicolor by PCR-RFLP,and to evaluate reliability of the approach as compared with biochemical classification.Methods Tween assimilation test and catalase reaction were carried out to identify 74 isolates of Malassezia species from patients with tinea versicolor and 7 Malassezia reference strains.The sequence of 28S rDNA of Malassezia species was amplified by PCR,and then the product was analyzed by RFLP with Eco88I,Bsp143Ⅱ and BshNⅠ,respectively.Results M.restricta,M.obtusa and M.pachydermatis were successfully identified by three restriction endonucleases.M.restricta was found to be more diverse from the other 6 species in genetic homology.By comparison with PCR-RFLP technique,a possible mistake was discovered with biochemical method.Conclusion PCR-RFLP is a promising molecular biological technique,which could rapidly and correctly classify Malassezia species.