[Clinical analysis of 33 cases of chronic liver diseases overlapping with CMV infection].

Objective: To understand the clinical features and outcomes of chronic liver diseases overlapping with CMV infection. Methods: Clinical characteristics, treatment and outcome of patients of chronic liver diseases overlapping with CMV infection were analyzed retrospectively. T-test was used for measurement data and χ (2) test was used for count data. All measurement data were expressed by (x ± s). P > 0.05 was not determined as significant. P < 0.05 was regarded as statistically significant. Results: Chronic liver diseases overlapping with CMV infections had similar clinical features. Etiopathogenic treatment + symptomatic supportive treatment + CMV overlapping infection treatment (including antiviral therapy, corticosteroids consideration, clearing heat and traditional Chinese choleretic medicine, etc) were the primary principles of therapy. The incidence of cytomegalovirus infection accounted for 4.125% during the corresponding hospitalization period. Cytomegalovirus infection had relatively caused liver function damage in patients with milder clinical symptoms and signs. Biochemical indicators before and after treatment showed that there was no significant difference in total bilirubin (TBil) before (262.93 ± 178.944) µmol/L and after one week of treatment (245.08 ± 179.332) µmol/L (P > 0.05). However, when TBIL was compared with three (156.58 ± 147.461) µmol/L and four weeks (103.39 ± 102.218) µmol/L) of treatment, the decrease was significant (P < 0.05, P < 0.01). Alanine aminotransferase (ALT) after one week (293.57 ± 467.438) U/L (P < 0.01) of treatment was significantly lower than before treatment (782.34 ± 828.801) U/L. Gamma-glutamyl transferase (GGT) after treatment (202.52 ± 155.174)U/L was significantly lower than before treatment(280.69 ± 205.619)U/L). Total bile acid (TBA) was increased after treatment (198.04 ± 155.174)µmol/L, when compared with that of before treatment (62.93 ± 178.944)µmol/L. Biochemical indicators of liver diseases had shown typical features of cholestasis, and the slow and reduced flow of bile acid was tracked and observed. Compared with the advanced group (182.45 ± 214.169) umol/L, the total bilirubin in inflammation group (50.36 ± 26.282) umol/L was decreased (P < 0.05). Moreover, advanced group (122.18 ± 106.780) umol/L (P < 0.05) had elevated total bile acid normalization rate than that of bile acid group (54.82 ± 56.123) umol/L, and the inflammatory phase had significantly better outcome than those with advanced-stage. Conclusion: Chronic liver diseases overlapping with cytomegalovirus infection has a good therapeutic outcome in the inflammatory phase, but in the advanced-stage; the therapeutic efficacy and outcome is poor and perilous.

Shared genetic etiology of hypertension and stroke: evidence from bioinformatics analysis of genome-wide association studies.

Hypertension is the most significant modifiable risk factor for cerebrovascular disease. It has been estimated that about 54% of strokes worldwide can be attributed to hypertension. However, there has not been a systematic study assessing the shared genetic susceptibility to hypertension and stroke on a genome-wide level. In this study, SNPs associated with essential hypertension and stroke were collected from the NHGRI-EBI GWAS catalog, and genotype imputation were conducted using information from the 1000 Genomes Project. Subsequently, the SNPs and the mapped genes were compared between the two diseases. Finally, functional clustering was performed, and the enriched GO terms and KEGG pathways were further compared between hypertension and stroke. Comparison of these two groups of SNPs and genes identified only one shared SNP (rs3184504) and 11 shared genes. After genotype imputation, 129 shared SNPs and 16 shared genes were identified. These genes were significantly enriched in 10 GO terms, which were mainly involved in lipoprotein and triglyceride metabolism. Additionally, KEGG analysis identified one pathway, glycerolipid metabolism, as being significantly enriched in both diseases. The present study strongly suggests that the gene network regulating lipid metabolism and blood circulation is the major shared genetic etiology of hypertension and stroke.

Dexmedetomidine increases acetylation level of histone through ERK1/2 pathway in dopamine neuron.

Dexmedetomidine is a highly selective α2-adrenoceptor agonist with sedation, anesthetic sparing, analgesia, sympatholytic, and neuroprotective properties. This study evaluated neuroprotective effects of dexmedetomidine on dopamine neurons correlated to histone acetylation via extracellular signal-regulated protein kinase 1 and 2 (ERK1/2) pathway. Animals were randomly assigned to four groups and treatments were given as onetime doses: dimethyl sulfoxide (DMSO; n = 6), dexmedetomidine 1 mg/kg ( n = 6), 10 mg/kg ( n = 6), and 100 mg/kg ( n = 6). Acetylation histone protein levels and ERK protein levels in rats dopamine neuron from striatum were determined by Western blotting after various doses of dexmedetomidine (1, 10, and 100 mg/kg) treatments. The messenger RNA expression related to signal transduction coupled to 5-hydroxytryptamine receptor (5-HTR) in striatum was assessed by quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Dexmedetomidine administration increased expression of ERK1/2 phosphorylation and histones H3 acetylation. PD098059, an inhibitor of pERK1/2, almost completely blocked dexmedetomidine-induced histones H3 acetylation. In addition, bioinformatics analysis in combination with qRT-PCR demonstrated that dexmedetomidine could regulate the genes that are related to signal transduction coupled to 5-HTR via α2-adrenoceptor. Our results define dexmedetomidine as a modulator of histones H3 acetylation via ERK1/2 signaling pathway in dopamine neuron from striatum, which may provide clues for the mechanism underlying the neuroprotective effects of dexmedetomidine.

Pathogenicity and distribution of highly pathogenic porcine reproductive and respiratory syndrome virus in pigs.

The pathogenesis of highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) strain (HuN4) is poorly understood. Therefore, highly pathogenic PRRSV strain (HuN4) and its derivative strain (HuN4-F112) (obtained by propagation in MARC145 cells for 112 passages) were inoculated into a total of 48 PRRSV-sero-negative pigs (age: 4-5 weeks) by the intranasal route. Virological, pathological and in situ hybridization analyses were performed. The results exhibited that pigs infected with HuN4 showed a loss of appetite, decrease in body weight, raised body temperature and respiratory symptoms, along with interstitial pneumonia lesions. In the HuN4 group, multifocal interstitial pneumonia with macrophage infiltration was found in the lung. The lesions in the lymph node were characterized by collapsed follicles, depletion of germinal centres and reduction in lymphocytes. Perivascular cuffing and glial nodules were observed in the brains of some pigs. By comparison, the HuN4-F112 group had milder lesions. PRRSV was detected in macrophages, alveolar epithelial cells and vascular endothelial cells in the tonsil and lymph nodes. The PRRSV amounts in the pigs infected with HuN4 were 10(5) -10(9) copies/ml in the blood and 10(10) -10(11) copies/g in the lung tissues, whereas the virus amounts with HuN4-F112 were 10(2.15) -10(3.13) copies/ml in the blood and 10(3.0) -10(3.6) copies/g in the lung. Our results demonstrate that the PRRS HuN4 virus infects alveolar epithelial cells, macrophages and vascular endothelial cells causing diffuse alveolar damage and lymph node necrosis. Its higher pathogenicity compared with HuN4-F112 virus may be explained in part by higher replication rate in the previously mentioned organs.

Genetic polymorphism of 12 Y-chromosomal STR loci in the Minnan Han Chinese in Southeast China.

The 12Y-chromosomal short tandem repeat (STR) loci DYS19, DYS389I and II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS385a and b were analyzed in a sample of 109 unrelated male individuals from mountainous areas of Minnan, Southeast China. A total of 95 haplotypes were identified. The allele diversity values for each locus ranged from 0.3205 (DYS391) to 0.9553 (DYS385), the haplotype diversity was 0.9863, and the discrimination capacity 0.8716.

The role of high-frequency jet ventilation in the treatment of Cheyne-Stokes respiration in patients with chronic heart failure.

BACKGROUND: Cheyne-Stokes respiration with central sleep apnea (CSR-CSA) is common in patients with severe cardiac failure. Various modes of positive airway pressure have been suggested as treatments. The authors hypothesized that high frequency jet ventilation (HFJV) can improve central sleep apnea in patients with chronic heart failure. METHODS: Eleven subjects with stable, optimally treated chronic heart failure and Cheyne-Stokes respiration were tested untreated and on four treatment nights in random order: nasal oxygen (4 L/min), continuous positive airway pressure (CPAP) (mean 9.3 cm H(2)O), bilevel positive airway pressure (BiPAP)(mean 12.0/5.3 cm H(2)O), or HFJV(60 approximately 80 breaths per minute) during polysomnography (EMBLA, Flaga, Reykjavik, Iceland). RESULTS: The apnea-hypopnea index (AHI) declined from 30.9 +/- 8.3/h in untreated night to 23.6 +/- 6.6/h oxygen night and 18.5 +/- 5.0/h CPAP, 14.3 +/- 3.9/h BiPAP, and 20.1 +/- 4.1/h HFJV (all P < 0.001 versus control). There was no significant difference between the AHI of HFJV and that of CPAP (P = 0.541). Arousal index decreased from 31.4+/-13.2/h untreated to 25.0 +/- 7.1/h oxygen and 13.6 +/- 4.7/h CPAP, to 13.7 +/- 4.9/h BiPAP and 14.4 +/- 4.7/h HFJV. HFJV had the similar effect to the other therapeutic groups in arousal index (P > 0.05). There were large increases in slow-wave and rapid eye movement (REM) sleep with HFJV. All subjects preferred HFJV to CPAP. CONCLUSIONS: One night of therapy with HFJV improved nocturnal breathing pattern and sleep quality in patients with Cheyne-Stokes respiration in chronic heart failure. HFJV therapy for sleep and breathing were the same as those during a nasal CPAP night. A long-term study of the effect of HFJV on cardiovascular function is needed.

Grain yield responses to moisture regimes in a rice population: association among traits and genetic markers.

Drought is a major constraint to rice (Oryza sativa L.) production in rainfed and poorly irrigated environments. Identifying genomic regions influencing the response of yield and its components to water deficits will aid our understanding of the genetic mechanism of drought tolerance (DT) of rice and the development of DT varieties. Grain yield (GY) and its components of a recombinant inbred population developed from a lowland rice and an upland rice were investigated under different water levels in 2003 and 2004 in a rainout DT screening facility. Correlation and path analysis indicated that spikelet fertility (SF) was particularly important for grain yield with direct effect (P=0.60) under drought stress, while spikelet number per panicle (SN) contributed the most to grain yield (P=0.41) under well-watered condition. A total of 32 quantitative trait loci (QTLs) for grain yield and its components were identified. The phenotypic variation explained by individual QTLs varied from 1.29% to 14.76%. Several main effect QTLs affecting SF, 1,000-grain weight (TGW), panicle number (PN), and SN were mapped to the same regions on chromosome 4 and 8. These QTLs were detected consistently across 2 years and under both water levels in this study. Several digenic interactions among yield components were also detected. The identification of genomic regions associated with GY and its components under stress will be useful to improve drought tolerance of rice by marker-aided approaches.

Effect of one-step 100% ethanol fixation and modified manual microdissection on high-quality RNA recovery from esophageal carcinoma specimen.

SUMMARY: This paper attempts to determine an optimal fixation protocol for stabilizing RNA during microdissection so as to obtain high-quality RNA from specific cell populations procured from esophageal carcinoma specimens, and to develop a manual microdissection that can facilitate the procurement. The special features of our protocol include one-step dehydration of tissue sections in 100% ethanol immediately after cryosectioning, a self-made T-shape plate (T plate) and "exclusion microdissection" procedure. The quality of RNA isolated from dissected cells was analyzed by neutral agarose gel electrophoresis and reverse transcription-polymerase chain reaction (RT-PCR) to detect genes of different abundance levels. One-step 100% ethanol fixation of cryosections effectively stabilized RNA integrity for agelong period of time while maintaining histological morphology comparable to that using the conventional procedure, indicating that it is a valid protocol for preservation of RNA in microdissected samples. In conjunction with the application of the T plate and 'exclusion microdissection' procedure, which efficiently simplifies manual microdissection procedure, allowing maximal procurement of target cells from complex primary tissues, full use of every single specimen for maximal procurement of target cells from the sections was allowed. The RNA isolated from 5 different stage-specific cell populations of an esophageal carcinoma specimen was of high quality and sufficient in quantity for various downstream molecular analyses. Our method is suitable for a wide spectrum of expression analysis in diverse clinical settings.

Genetic profile of 15 STR loci in the Min Nan population in Southeast China.

Genetic profile of 15 short tandem repeats (STR) loci were determined in a Chinese Han population from the Min Nan mountainous area, Southeast China.

Further evidence for an HLA-related recessive mutation in nasopharyngeal carcinoma among the Chinese.

We typed 247 cases of nasopharyngeal carcinoma (NPC), a disease predominantly of the southern Chinese, and 274 controls from the Chao Shan region of China's Guangdong province for HLA A and B. Besides confirming the established associations with A2, A33, B46 and B58 (positive associations) and A11 (negative association), the results demonstrated a number of rarer alleles with strong negative association with NPC. Our data, combined with those from the previous studies in Southern Chinese, displayed the protective effects for A31 (odds ratio (OR)=0.0; 95% confidence interval (CI)=0-0.11), B13 (OR=0.50; 95% CI=0.35-0.69), B27 (OR=0.49; 95% CI=0.25-0.92), B39 (OR=0.18; 95% CI=0.06-0.48) and B55 (OR=0.32; 95% CI=0.14-0.68), the ORs comparing individuals with or without each allele. Other ethnic groups do not display such large HLA-associated variation in NPC risk. We show that a linked NPC gene with dominant mode of action could not generate such large protective effects. The results provide strong supporting evidence for the existence of a southern Chinese specific, recessive NPC gene closely linked to the HLA region as a major determinant of the Chinese risk for the disease.

Analysis of STR polymorphisms in the Chao Shan population in South China.

Chao Shao area is a littoral under the jurisdiction of Guangdong province, abutting on Fujian. Historically, the area was relatively isolated from other parts of China until 1990s, when it started to take a small number of immigrants from other regions. People residing in this area speak in unique dialect and have distinct lifestyle. Allele frequencies for the 15 short tandem repeats (STR) loci included in the AmpFLSTR Identifiler kit were obtained from a sample of 144 unrelated Chinese born and living in the Chao Shan area, South China.

Lysine-derivatized polyurethane as a clot lysing surface: conversion of adsorbed plasminogen to plasmin and clot lysis in vitro.

Polyurethane surfaces to which lysine residues are immobilized by photochemical methods are proposed as a basis for clot lysing surfaces. The lysines are attached in such a way that the epsilon-amino and carboxyl groups are free. We showed previously that these surfaces, when placed in contact with plasma, adsorb only plasminogen and virtually no other proteins (McClung et al., J. Biomed. Mater. Res. 49 (2000) 409). In this communication, data based on a chromogenic substrate assay are presented showing that plasminogen adsorbed to these surfaces is readily converted to plasmin in the presence of tissue-plasminogen activator (t-PA). Moreover, the rate of activation on the surface is considerably greater than in solution. Experiments demonstrating the ability of these surfaces to dissolve fibrin clots are also reported. Surfaces exposed to plasma and then to t-PA were placed in citrated plasma. On recalcification, clotting was initiated, but the incipient clots were soon dissolved. On control surfaces (no lysine or lysine in which the epsilon-amino groups were not available) coagulation continued until a stable clot was formed. Similar observations were made when the plasma/t-PA exposed surfaces were placed in a pure fibrinogen solution and thrombin was added.

Adsorption of plasminogen from human plasma to lysine-containing surfaces.

The objective of this work is to develop blood-contacting surfaces that will dissolve nascent clots that may begin to form on them. Surfaces were prepared consisting of a polyurethane to which a coating reagent was attached covalently by photochemical methods. The coating reagent was a polyacrylamide with lysine and benzophenone (for photochemical attachment) moieties pendant to the chains. It was hypothesized that via the lysine moieties such surfaces would show specific binding affinity for plasminogen, the principal component of the fibrinolytic system in blood. Surfaces of varying lysine content in which the lysine was bound through the alpha-amino groups, leaving the epsilon-amino groups free, were investigated. A control surface in which the lysine was bound through the epsilon-amino groups was also examined. Advancing water contact angles showed the surfaces to be hydrophilic. Hydrophilicity was found to decrease as the lysine content increased. Adsorption of plasminogen from plasma was studied using radioiodinated plasminogen as a tracer. For the epsilon-lysine surfaces, adsorption increased with increasing lysine content and reached a value of 1.2 microg/cm(2) for the surface with the highest lysine content, that is, in the range expected for a compact monolayer of plasminogen. The control surfaces, which contained either no lysine or lysine in which the epsilon-amino groups were unavailable, adsorbed very small amounts of plasminogen. Immunoblots were obtained for the proteins eluted from the surfaces after incubation with plasma. For the control surfaces, most of the proteins tested for (some 20 in all) were present. However, for the surface containing the highest concentration of epsilon-lysine, only plasminogen was detected in a significant amount. It is concluded that the epsilon-lysine surface adsorbs plasminogen to the exclusion of the other plasma proteins. Studies to examine the fibrinolytic properties of these surfaces will constitute the next phase of this work.

Surface texturing and coating of biomaterial implants: effects on tissue integration and fibrosis.

To determine whether texturing and coating have additive effects in promoting tissue integration and inhibiting fibrosis, we evaluated smooth silicone rubber (SSR), textured silicone rubber (TSR), porous silicone rubber (PSR), expanded polytetrafluoroethylene (ePTFE), and porous polyurethane (PPU) subcutaneous implants in eight minipigs. Some of the implants were coated with type IV collagen (Col) and/or fibronectin (Fn). At 6 months, we removed the implants and examined them microscopically. Texturing was more important than Col and Fn in reducing fibrosis and inflammation. The PSR yielded the best response, including reduced fibrosis and inflammation, satisfactory adherence, and no dystrophic mineralization.

Nutrition knowledge, attitudes and practices among senior medical students in Taiwan.

OBJECTIVE: Nutrition is an important issue in medical training, but the nutritional knowledge, attitudes and practices of medical students in Taiwan have not been elucidated. METHODS: This investigation was a need assessment that examined knowledge, attitude and practices of medical students in selected areas of nutrition. A national sample of 528 senior medical students from nine medical colleges in Taiwan participated in this study by completing a questionnaire. RESULTS: On a 10-point scale, the average score of students on general and clinical nutritional knowledge was 5.99 +/- 1.51 and 5.15 +/- 1.77, respectively. The percentage of correct answers from questionnaires in both areas was 60% and 52%, respectively. Seventy-seven percent or more of the students reported that they either agreed or strongly agreed with four positive-attitude statements and either disagreed or strongly disagreed with two out of three negative-attitude statements. Between 30% and 61% of the students reported that they practice on nutrition-related individual behaviors. CONCLUSIONS: The knowledge, attitudes and practices of senior medical students in Taiwan suggest the need for education strategies to improve competence in the area of nutrition.

PIP: A questionnaire administered to 528 senior medical students from all 9 medical colleges in Taiwan revealed a need for curriculum modifications to improve nutrition-related knowledge, attitudes, and practices. At present, 5 of the medical schools offer elective courses on nutrition, but a nutrition curriculum is not required during medical training. The 20 items on the questionnaire concerned the nutritional functions of various nutrients, nutrition management in disease states, nutrition in disease prevention, and nutrition status assessment. On a 10-point scale, the average score was 5.99 on general nutrition knowledge and 5.15 on clinical nutritional knowledge. Correct responses were highest (77.0%) on the 2 questions concerning the nutritive content of foods and lowest (17.35%) on nutrition status assessment. Only 50% knew the definition of a balanced diet and just 30% were concerned about the caloric content of their own diet. Overall, these findings suggest that nutrition education, including an evaluation of one's own diet, should be incorporated into the training programs of Taiwanese medical students.

Nutrition knowledge, attitude and practice among primary care physicians in Taiwan.

OBJECTIVE: To investigate nutritional knowledge, attitude and practices among primary care physicians in the Taiwan area. METHOD: A closed-end questionnaire containing 26 knowledge questions, 12 attitude statements and 12 practice statements was mailed to physicians on the mailing list of the National Health Administration (NHA). RESULTS: The data reported are based on the responses of 27% of the total 1210 primary care physicians in the Taiwan area. Physicians answered 59% of the total knowledge questions correctly, with a tendency to score higher on general knowledge than clinical nutrition. The majority of physicians tended to agree with the positive-attitude statements and disagree with the negative-attitude statements. The performance of physicians regarding personal practices was less than that for job-related practices. CONCLUSIONS: This nationwide survey of nutrition-related knowledge and practices demonstrates the need for nutrition education for physicians. The questionnaire may be a useful instrument for future educational strategies in Taiwan.

PIP: A questionnaire completed by 331 primary health care physicians in Taiwan revealed deficiencies in nutrition-related knowledge, attitudes, and practices. Questionnaires were sent to all 1210 physicians on the mailing list of the National Health Administration; the response rate was 27%. Physicians answered 59% of the 26 knowledge-related questions correctly. The highest proportion of correct responses was obtained for questions related to nutrient functions and nutrition during pregnancy (both 70.6%), while the lowest was recorded for the item concerning nutritional assessment (42%). Overall, physicians considered nutrition to be important in their personal and clinical practice. However, only 78% expressed agreement with the statement that nutrition consultation should be a part of health care. Nutrition knowledge was higher among female physicians, those under 35 years of age, and non-smokers. Finally, there was a significant correlation between nutrition knowledge and attitudes. Although this study is limited by the poor response rate, the results indicate a need for improvements in the basic nutritional knowledge and practices of primary care physicians in Taiwan.

Induction of the chemotactic S100 protein, CP-10, in monocyte/macrophages by lipopolysaccharide.

The murine S100 protein CP-10 is a potent chemotactic factor for murine and human myeloid cells in vivo and in vitro. This is the first report describing regulations of the CP-10 gene by a proinflammatory stimulus, lipopolysaccharide (LPS), in cells of the monocyte/macrophage lineage. Murine monocyte/macrophage-like WEHI 265 and RAW 264.7 cells preexposed to 5 to 50 ng/mL LPS expressed significant levels of CP-10 mRNA 4 hours, and maximal at 20 hours, after a secondary LPS challenge. This was accompanied by increasing levels of cell-associated and released CP-10 protein. In contrast, a single dose of LPS upregulated CP-10 mRNA in elicited peritoneal macrophages, whereas mRNA and protein levels decreased following LPS challenge. The state of macrophage differentiation may control responsiveness as LPS had no effect on CP-10 basal levels in bone marrow derived macrophages. LPS-induced CP-10 expression was controlled at the transcriptional level and nuclear run-on and protein synthesis inhibition assays indicated that LPS priming and challenge of RAW cells occurred via distinct pathways. MRP14, another S100 protein generally coordinately expressed with human MRP8, was not induced by LPS under the same conditions. We propose that CP-10 may play a key role in recruitment of leukocytes into tissues in response to gram-negative bacterial infection.

Polymerase chain reaction (PCR) mutagenesis enabling rapid non-radioactive detection of common beta-thalassaemia mutations in Mediterraneans.

The IVS-1-110 (G----A) and IVS-1-1 (G----A) mutations occur in approximately 33% and 9% respectively of beta-thalassaemia alleles in Mediterraneans (Kazazian & Boehm, 1988). They are generally detected in polymerase chain reaction (PCR)-amplified material by allele-specific oligonucleotide (ASO) hybridization patterns. In this study, artificial base substitutions in amplified material have been created to distinguish normal from mutant alleles on the basis of restriction enzyme digestion patterns. Invariant target sites provide an internal control for restriction enzyme activity. Mutagenesis was achieved by 3' base mismatches in primers selected to anneal immediately adjacent to target sites. Digestion of PCR products from normal and thalassaemic alleles with the restriction enzymes MboI (IVS-1-110) and HinfI (IVS-1-1) produced different fragments on electrophoresis. The above strategy was validated by allele-specific oligonucleotide probing. Identification of the three commonest mutations in this population (IVS-1-110, codon 39 and IVS-1-1), which account for approximately 69% of thalassaemic alleles (Kazazian & Boehm, 1988), was subsequently undertaken in seven chorion villus biopsies.

Utility of the polymerase chain reaction (PCR) for prenatal diagnosis of genetic disease.

Gene amplification by the polymerase chain reaction (PCR) has been applied to prenatal diagnosis for alpha and beta thalassemias (1 and 5 cases respectively), Hemoglobin (Hb) Lepore/beta thalassemia (1 case) and cystic fibrosis (14 cases). Chorionic villus samples were obtained in the tenth week of pregnancy and DNA analysed in parallel with conventional gene mapping. Direct diagnosis of the common Mediterranean beta-thalassemia mutations (IVS-1-110 and codon 39), Hb Lepore, and the delta F508 mutation causing cystic fibrosis was achieved by hybridization of amplified material with pairs of allele-specific oligonucleotide (ASO) probes or by restriction enzyme digestion of PCR products. Results were confirmed by DNA mapping. Definitive diagnosis or exclusion of an affected fetus was possible in 17 of 21 cases thus examined. PCR reduces the time required for prenatal diagnosis. DNA contamination is a potential source of error.