Two distinct phenotypes of triploid fetuses have been previously described and a correlation with parental origin of the triploidy has been suggested. We have studied the parental origin of the extra haploid set of chromosomes in nine triploid fetuses using analysis of DNA polymorphisms at a variety of loci. Maternal origin of the triploidy (digyny) was demonstrated in six fetuses with type II phenotype, paternal origin (diandry) in two cases with type I phenotype, and nonpaternity in one case. The predominance of digynic triploids in our study contrasts with the results reported in previous studies in which, through analysis of cytogenetic polymorphisms, paternal origin was found to account for the majority of triploid conceptuses. This difference may be accounted for by a combination of factors--the different methods of parental assignment used and analysis of a different subset of triploid conceptuses. The correlation between the observed phenotypes and the parental origin of triploidy may represent another example of imprinting in human development.

Kallistatin is a serine protease inhibitor with anti-inflammatory, anti-angiogenic, and anti-oxidative properties. Since oxidative stress plays a critical role in the pathogenesis of diabetic nephropathy, we studied the effect and mechanisms of action of kallistatin superinduction. Using ultrasound-microbubble-mediated gene transfer, kallistatin overexpression was induced in kidney tubules. In db/db mice, kallistatin overexpression reduced serum creatinine and BUN levels, ameliorated glomerulosclerosis and tubulointerstitial injury, and attenuated renal fibrosis by inhibiting TGF-β signaling. Additionally, downstream PAI-1 and collagens I and IV expression were reduced and kallistatin partially suppressed renal inflammation by inhibiting NF-κB signaling and decreasing tissue kallikrein activity. Kallistatin lowered blood pressure and attenuated oxidative stress as evidenced by suppressed levels of NADPH oxidase 4, and oxidative markers (nitrotyrosine, 8-hydroxydeoxyguanosine, and malondialdehyde) in diabetic renal tissue. Kallistatin also inhibited RAGE expression in the diabetic kidney and AGE-stimulated cultured proximal tubular cells. Reduced AGE-induced reactive oxygen species generation reflected an anti-oxidative mechanism via the AGE-RAGE-reactive oxygen species axis. These results indicate a renoprotective role of kallistatin against diabetic nephropathy by multiple mechanisms including suppression of oxidative stress, anti-fibrotic and anti-inflammatory actions, and blood pressure lowering.

The EGFR array provides a sensitive, inexpensive, and robust method for monitoring non-small-cell lung cancer patients' response to TKI, and obviates the need of repeated lung biopsy.

Summary.A reverse dot blot method based on membranebound allele-specific oligonucleotides as hybridization targets for amplified a-gene fragments has been developed for the rapid detection of four non-deletion a thalassaemia defects found in the Chinese. Since these non-deletion defects account for 22·8% of haemoglobin H disease, a sensitive, specific and rapid screening method should be of value.Keywords: reverse dot blot, allele-specific hybridization, nondeletion a thalassaemia.Alpha thalassaemia (thal) is a group of hereditary anaemias which are commonly due to deletion of one or more of the aglobin genes on chromosome 16. In humans the a genes are duplicated and loss of one and two a genes on the same chromosome (a thal 2 and a thal 1 phenotype respectively) do not manifest any clinical symptoms. Loss of three genes (Hb H disease) results in a thal intermedia picture, and complete deletion of all four a-globin genes (Hb Barts hydrops foetalis) results in severe anaemia in utero. Although most cases of Hb H disease are due to gene deletion, it can also occur as a result of compound heterozygosity of a thal 1 and a non-deletion a thal (a T a). Such cases usually result in a more severe phenotype, some with severe iron overload, despite infrequent transfusion episodes (Todd, 1971;Chim et al, 1998). In a recent study of 114 Hb H patients the prevalence of such non-deletion cases accounted for up to 22·8% (unpublished observations). In South-East Asia four major types of non-deletion a thal are found, viz a2 codon 30 (DGAG), a2 codon 59 (G → A), a Quong Sze codon 125 (T → C) and a Constant Spring codon 142 (T → C). Since it is now possible to selectively amplify the two a genes (Dodé et al, 1990;Hall et al, 1993), we have devised a reverse dot blot allele-specific hybridization method for the rapid detection of the four types of non-deletion defects which will circumvent the need for direct genomic sequencing. MATERIALS AND METHODS150 ng of genomic DNA was used for selective PCR amplification of the a2 genes. A 5 0 primer C1 (nt 6464-6483) (nucleotide numbering according to Humhba4, GenBank/EMBL database J00153) and an a2-specific 3 0 primer C3 (nt 7548-7529) were used. Amplification was made using conditions as previously described (Chan et al, 1997) which generated an a2 gene fragment of 1085 basepairs (bp).For more specific amplification, a semi-nested second PCR (30 cycles) was made using an internal 5 0 primer 501 (nt 6692-6712) with C3, 2 ml of the first PCR product as template and the addition of 750 pmol biotin-dUTP (BRL, U.S.A.). The reactants and conditions were the same as before.Preparation of membrane strips. Biodyne C (Pall Biomedical, U.S.A.) membrane was activated briefly in 0·1 N HCl, rinsed with water and soaked in 16% 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide (EDC) for 15 min; it was rinsed in water and air dried overnight. Oligonucleotide probes were diluted with 0·5 M NaHCO 3 /Na 2 CO 3 buffer, pH 8·4, to the desired concentration (ϳ5 pmol/ml) for application onto the membrane. Approxim...