Faculty of Medicine, Ege University, Bornova, Izmir, Turkey, and 157 healthy individuals were studied. Pa­tients with common bile duct stone and cholangitis did not have pancreatitis on the basis of radiological, laboratory and clinical tests. All patients and healthy control individuals were over 18 years of age, were informed about the study and consented to take part; they also consented to be recalled after 4 years.

All patients were checked for fasting blood sug­ar, blood urea, creatinine, serum alanine aminotrans-ferase (ALT), serum aspartate aminotransferase (AST), serum alkaline phosphatase, gamma-glutam-yltransferase (GGT), serum amylase, serum lipase, serum lactate dehydrogenase (LDH), sodium (Na), potassium (K), calcium (Ca), serum total cholesterol, trigylcerides, serum high density lipoprotein (HDL), serum low-density lipoprotein (LDL) and serum very low density lipoprotein (VLDL) levels. Complete blood counts (CBCs), whole abdominal ultrasonog-raphy and abdominal tomography were performed on all patients. All patients had abdominal pain, fe­ver, raised ALT, AST and serum alkaline phosphatase levels, whereas pancreatitis patients only had elevat­ed amylase levels. The serum bilirubin level was above 2 mg/dL in all patients. Endoscopic retrograde cholangiopancreatography (ERCP) and sphincterec-tomy were performed on all patients.

The cholangitis and biliary pancreatitis patients were recalled 4 years later and their prognosis was evaluated. For detection of the ACE polymorphism, blood specimens from all participants were obtained by standard venipuncture into ethylenediamine tetra-acetic acid-containing tubes. DNA was isolated by a standard phenol/chloroform extraction method [14].

We used the polymerase chain reaction (PCR) methodology [19] with an upstream primer (5'-CTG GAG ACC ACT CCC ATC CTT TCT-3') and a down­stream primer (5'-GAT GTG GCC ATC ACA TTC GTC AGA T-3'). Amplification was performed for 35 cycles with denaturation, extension and annealing temperatures of 94.8°C, 60.8°C and 72.8°C, respec­tively. The resulting PCR products were separated on 2% agarose gel with ethidium bromide staining and visualized under ultraviolet light. Homozygotes for the deletion or insertion genotypes were described as DD and II, respectively, and the heterozygotes were reported as ID.

All statistical analyses were performed using the SPSS 11.0 statistical program. Genotypes, allele frequency, and clinical features at diagnosis were evalu­ated by the x2 test. Clinical data are reported as mean ± SD (standard deviation) and as percentages. Statis­tical significance was accepted as p <0.05.