SNP and mRNA expression for glutathione peroxidase 4 in Kashin-Beck disease.

Kashin-Beck disease (KBD) is a chronic endemic osteoarthropathy, which mainly occurs in West and Northeast China. Epidemiological studies suggest that Se deficiency is an important environmental factor for the incidence of KBD. Glutathione peroxidase 4 (GPx4) belongs to the glutathione peroxidase family, which is crucial for optimal antioxidant defences. Our purpose is to investigate the putative association between GPx4 polymorphisms and the risk of KBD. Restriction fragment length polymorphism-PCR was used to detect two SNP (rs713041, rs4807542) in 219 cases and 194 controls in Han Chinese subjects, and quantitative analysis for the GPx4 mRNA level was performed by the real-time PCR method. The results revealed that linkage disequilibrium existed in the two SNP. A significant difference was observed in the haplotype A-T (P = 0·0066) of GPx4, which was obviously lower in the KBD cases (0·006 v. 0·032 %). Correlation analysis based on a single locus showed no association between each SNP and KBD risk. Furthermore, the GPx4 mRNA level was dramatically lower in the blood of KBD patients. Overall, our finding indicated GPx4 polymorphisms and decreased mRNA level may be related to the development of KBD in the Chinese population, suggesting GPx4 as a possible candidate susceptibility gene for KBD.

Expressions of nucleoside diphosphate kinase (nm23) in tumor tissues are related with metastasis and length of survival of patients with hepatocellular carcinoma.

OBJECTIVE: To evaluate the relationship of expressions of nucleoside diphosphate kinase (nm23) and proliferating cell nuclear antigen (PCNA), as well as apoptosis, with the prognosis of HCC patients by analyzing their pathological and clinical data. METHODS: The expressions of nm23 and PCNA were analyzed by immunohistochemistry and the apoptotic phenomena were detected by TUNEL technique in the liver samples from 43 HCC tissues, 39 para-neoplastic tissues, and 10 normal tissues. The mean apoptosis index (AI) and proliferative index (PI) in individual sample were calculated. RESULTS: As shown by the detection, 32.6% of carcinomas had negative nm23 signal in tumor tissues, whereas all para-neoplastic and normal tissues had positive nm23. The AI in nm23 positive HCC was significantly higher than that in nm23 negative one, with statistical difference (P<0.05). Furthermore, the expressions of nm23, and the values of AI and PI were contrastively analyzed with some main pathological and clinical data of HCC. It revealed that HCC with extrahepatic metastasis showed remarkable correlation with the negative nm23 (P=0.013) and higher PI values of HCC (P=0.015). The disease-free survival in HCC patients with negative nm23 expression was significantly poorer than that in patients with positive nm23 expression. CONCLUSIONS: These data suggest that expressions of nm23 protein in tumor tissues are correlated with occurrences of metastasis and length of survival of the HCC patients, which may be an indicator for their prognosis.

Over-expression of nm23-H1 in HeLa cells provides cells with higher resistance to oxidative stress possibly due to raising intracellular p53 and GPX1.

AIM: To determine whether the antitumor factor nm23 is related with antioxidation. METHODS: Full-length human nm23-H1 was cloned into a mammalianexpressing vector and transiently introduced into HeLa cells. RESULTS: A remarkably low level of reactive oxygen species (ROS) was detected in the cells overexpressing nm23-H1. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays found that the cells transfected with a nm23- H1-expressing plasmid had higher viability and stronger resistance to oxidative stress. Immunoprecipitation tests revealed that endogenous nm23-H1 formed a protein complex with p53. Furthermore, the intracellular levels of p53 and p53- regulated gene GPX1 were obviously increased in the cells overexpressing nm23- H1. The downregulation of p53 in the cells overexpressing nm23-H1 resulted in a higher cellular ROS level and lower cell viability. CONCLUSION: The findings suggest that nm23-H1 may act as a cellular protector against oxidative stress, possibly triggering the p53-related antioxidative pathway.

[Full length nucleotide sequence analysis of L and M segments of Hantavirus Q32 strain gene].

BACKGROUND: To study the full length L and M sequence of Hantavirus Q32 strain gene and explore its molecular characters. METHODS: The L and M segment cDNA of Hantavirus Q32 strain was amplified by RT-PCR. The purified PCR products were sequenced directly or cloned into pGEM-T Vector and then sequenced. RESULTS: The L genome segment of Q32 virus was found to be 6533 nucleotides in length. One large open reading frame was found located at bases 38 to 6493. This was predicted to encode an L protein 2151 amino acids in length with a molecular mass of 2.46 x 10(5). The M genome segment was 3616 nucleotides in length. One open reading frame was located at bases 41 to 3488. This was predicted to encode an M protein 1135 amino acids with a molecular mass of 1.26 x 10(5). CONCLUSION: The nucleotides sequence of M and L segments of strain Q32 was similar to that of other Hantavirus M and L segments. Deduced amino acid sequences of glycoprotein and RNA polymerase revealed high homologue to other Hantavirus.

[Dynamic effects of gallium chloride on osteoporotic rat model induced by tretinoin].

OBJECTIVE: To investigate the effect of gallium salts on bone metabolism in osteoporosis rats caused by tretinoin. METHODS: After duplicating osteoporotic model of rats by using tretinoin, we observed the metabolism of blood, urine and bone in vivo. The rats were divided into control group, osteoporosis group, estrogen treated and gallium chloride treated group. The indexes of bone metabolism and related indexes in serum and urine were observed after 60 days of treatment. In the same time, we observed the dynamic effect of 30 days and 60 days of treatment. RESULTS: After duplicating osteoporotic model, rats' bone structures were injured, the bone mineral density and the organic matrix decreased, the contents of tartrate-resistant acid phosphatase (TRAP) in serum were higher. After gallium salt treating, the above damages were inhibited or retarded. Trabecular width and thickness of bone cortex were significantly increased. AKP and TRAP activities were decreased to the level close to that of the control group. CONCLUSION: Gallium salt is effective in treating osteoporosis.

A dual-wavelength resonance light scattering ratiometry of biopolymer by its electrostatic interaction with surfactant.

A dual-wavelength resonance lighting scattering (DW-RLS) ratiometry is developed to detect anion biopolymer based on their bindings with cation surfactant. Using the interaction of Hyamine 1622 (HM) with fish sperm DNA (fsDNA) as an example, a dual-wavelength resonance light scattering (DW-RLS) ratiometric method of DNA was constructed. In Britton-Robinson buffer controlled medium, fish sperm DNA (fsDNA) could interact with Hyamine 1622 (HM), displaying significantly enhanced RLS signals. By measuring the RLS signals characterized at 300.0nm (I(300.0)) and the RLS intensity ratio (I(276.0)/I(294.0)), respectively, fsDNA over a wide dynamic range of content could be detected. Typically, when HM concentration is kept at 6.0x10(-5)moll(-1), using I(300.0) could detect fsDNA over the range of 50-2000ngml(-1) with the limit of 3.0ngml(-1), while using I(276.0)/I(294.0) could detect fsDNA over the range of 0.5-2500ngml(-1) with the limit of 0.05ngml(-1). Thus the latter so-called DW-RLS ratiometry is obviously superior to the former one. Based on the measurements of I(300.0) and I(276.0)/I(294.0) data, a Scatchard plot concerning the interaction between HM and fsDNA could be constructed and thus the binding number (n) and binding constant (K) could be available with the values of 13.5 and 1.35x10(5)mol(-1)l, and 11.9 and 1.65x10(5)mol(-1)l, respectively.