Telomerase activity in human hepatocellular carcinoma: parallel correlation with human telomerase reverse transcriptase (hTERT) mRNA isoform expression but not with cell cycle modulators or c-Myc expression.

BACKGROUND: To explore the possible regulatory mechanisms of telomerase, we examined the telomerase activity (TA), expression of human telomerase RNA (hTR), human telomerase reverse transcriptase (hTERT) mRNA isoforms and cell cycle modulators in human hepatocellular carcinoma (HCC) cell lines (J5, J7) and a normal human immortalized hepatic epithelial cell line (Chang-liver). METHODS: The cell lines were chemically synchronized in either G1, G1/S, G2/M or M phases. TA was measured by polymerase chain reaction (PCR)-based telomerase repeat amplification protocol assay. The hTR and hTERT mRNA levels were analyzed by reverse transcriptase-polymerase chain reaction. Western blotting and immunocytochemistry were used to assay the cell cycle modulators. RESULTS: The TA of J5, J7 and Chang-liver cell lines tested was highest in M phase. The expression level of hTERT mRNA associated with the highest TA detected in the M phase of HCC cell lines. Chang-liver expressed markedly less TA and hTERT mRNA than J5 or J7. The elevated TA and expression of hTERT mRNA isoforms in M phase of HCC cell lines did not significantly correlate with that of the cell cycle modulators and c-Myc. CONCLUSIONS: The results implicate that regulation of TA is related to hTERT mRNA isoform expression, and that regulation is different between the cell immortalization and tumorigenesis.

Polymorphism of the beta(2)-adrenoceptor in COPD in Chinese subjects.

STUDY OBJECTIVES: To assess the frequencies of three polymorphisms at amino acid positions 16, 27, and 164 of the beta(2)-adrenoceptor (beta(2)-AR) gene and their effects on COPD patients. DESIGN: Prospective, case-control study PATIENTS: Sixty-five patients with COPD and 41 healthy subjects were included. MEASUREMENTS: Polymorphisms of the beta(2)-AR coding block were delineated using an allele-specific polymerase chain reaction (PCR) approach. The allele-specific PCR technique was verified by direct dideoxy sequencing of PCR products. Pulmonary function tests were performed in all patients. RESULTS: The Arg16 beta(2)-AR polymorphism was less prevalent in COPD patients than in healthy populations (p = 0.01). A significant correlation (p < 0.018) between the Gln27 beta(2)-AR polymorphism and FEV(1) percent predicted value was found. Patients with the Gln27 polymorphism had a higher percentage of low FEV(1) percent predicted than did patients with the GlnGlu and GluGlu variants. CONCLUSIONS: The polymorphism of Gly16 may increase the patient's susceptibility to the development of COPD. The Gln27 beta(2)-AR polymorphism may be associated with the severity of COPD in a Chinese population.

Telomerase activity correlates with cell cycle regulators in human hepatocellular carcinoma.

AIMS/BACKGROUND: Mutation in cell cycle genes is the most common genetic change in malignant tumor cells. Telomerase activation, considered as essential in the immortality of cancer cells, is found in most cancers, where there may be an association with an active cell cycle. METHODS: In this study study we used the TRAP assay to determine telomerase activity in liver tumor specimens from 25 cases of hepatocellular carcinoma (HCCs) as well as in corresponding non-cancerous liver tissue in each patient. The expression of cyclin D1, cdk2, and cdk4 protein was also examined by Western blot. RESULTS: Twenty-one of the 25 cases of HCC were found to have increased telomerase activity, whereas only five out of the 25 non-cancerous liver samples were found to have weak telomerase activity. Telomerase activity was not found to be related to tumor size, HBsAg, HBeAg, anti-HCV, transaminase, or alpha-fetoprotein serum titer. Furthermore, three out of the 25 cases of HCC showed cyclin D1 overexpression, whereas 15 of the 23 cases of HCC showed decreased cyclin D1 expression. Down regulation of cyclin D1, cdk2, cdk4 protein correlated with telomerase activity (p<0.004, p<0.013, and p<0.001 respectively). CONCLUSION: The results indicate that genetic defects in HCC facilitate the reactivation of telomerase activity, a process which may be dependent on cyclin D1 with its cyclin dependent kinase (cdk) partner defect.

Molecular studies into the role of CD44 variants in metastasis in gastric cancer.

CD44, an integral membrane glycoprotein expressed by many cell types, serves as the principal transmembrane hyaluronate receptor and might be a determinant of metastatic and invasive behaviour in carcinomas. The generation of CD44 splice variants might be linked closely with gastric carcinoma tumorigenesis and differentiation. Some studies have reported that the magnitude of CD44 variant synthesis at the protein level correlates with lymph node metastasis. A number of studies have examined the possible mechanism of involvement of the CD44 variant in tumour metastasis. Most studies have reported that the regulation of CD44 binding to hyaluronate results from glycosylation of variably spliced exons. Direct hyaluronate binding studies of CD44 V4-V7 isoforms transfected into the human gastric carcinoma cell line, SC-M1, have indicated that the V4-V7 isoforms themselves, in addition to glycosylation, can alter hyaluronate binding.

Apoptosis in nasopharyngeal carcinoma as related to histopathological characteristics and clinical stage.

AIMS: We investigated the significance of apoptosis, using the terminal deoxynucleotidyl transferase mediated dUTP-digoxigenin nick end-labelling method, in nasopharyngeal carcinoma biopsy samples. METHODS AND RESULTS: The apoptotic index (AI) in 50 nasopharyngeal carcinomas was compared with various histopathological features and clinical stage. Also, the AI was correlated with p53, bcl-2 and Ki67 expression by immunohistochemistry. In histopathological studies, the AI was significantly higher in mixed cellular type (MC) than in keratizing squamous cell type (KS) and spindle cell type (SC) (P < 0.001) which worsens prognosis. In tumour stage analyses, AI was higher in early stage (stage 2 and 3) than in high stage (stage 4). In addition, there was a significant correlation between the AI and p53 expression (P < 0.001) but not with proliferative activity (P = 0.15). In NPC containing p53 protein positive tumour cells, there was a significantly higher apoptotic rate. CONCLUSIONS: These findings indicate that apoptosis is related to type and stage of nasopharyngeal carcinoma. They also confirm the role of p53 in regulating tumour apoptosis.

The extent of proliferative and apoptotic activity in intraductal and invasive ductal breast carcinomas detected by Ki-67 labeling and terminal deoxynucleotidyl transferase-mediated digoxigenin-11-dUTP nick end labeling.

BACKGROUND: The balance among cell proliferation, cell differentiation, and cell death determines the cell number in a population as well as the size or even the stage of a tumor. Thus, to improve our understanding of the pathogenesis of neoplasms, it is important to investigate the regulation of both cell proliferation and cell death. METHODS: This study examined the occurrence of apoptosis and proliferative capacity in 46 breast carcinomas: 20 intraductal carcinomas (ductal carcinomas in situ [DCIS]) and 26 infiltrative ductal carcinomas (IDC). Terminal deoxynucleotidyl transferase-mediated digoxigenin-11-dUTP nick end labeling (TUNEL) and immunostaining with the Ki-67 antibody were used in the examination. A ladder of DNA fragments induced by apoptosis was demonstrated by means of DNA agarose gel electrophoresis in 10 of the available TUNEL positive and negative samples. RESULTS: The results were correlated with p53, bcl-2, estrogen receptor (ER), and progesterone receptor (PR) protein expression, which would suggest association with apoptosis by immunohistochemistry. The apoptosis and proliferation of each cancer were expressed as the number of tumor cells undergoing apoptosis and proliferation per 1000 tumor cells. The extent of apoptosis was more frequently observed in DCIS than in IDC (21.9+/-6.8 vs. 4.0+/-0.9, P < 0.001), and the proliferation activity was significantly higher in IDC than in DCIS (16.8+/-6.5 vs. 3.5+/-0.8, P < 0.006). Apoptosis associated with MIB-1 positive cells and TUNEL labeling was significantly higher in IDC than in DCIS (3.26 vs. 0.42, P=0.001). In DCIS, apoptosis was correlated with p53 (r=0.663, P=0.005), and p53 had a reverse correlation with bcl-2 (r=0.620, P= 0.018). Moreover, bcl-2 expression was associated with ER (P=0.028) and PR (P= 0.005) expression in both DCIS and IDC. CONCLUSIONS: The results of this study show that a higher degree of apoptosis and lower proliferation activity in intraductal carcinoma result in a steady-state, self-renewing condition in which net growth of the tumor is rare. The results also indicate that apoptosis was altered by the expression of p53, bcl-2, ER, and PR.

Hyaluronate binding assay study of transfected CD44 V4-V7 isoforms into the human gastric carcinoma cell line SC-M1.

The potential human metastasis molecule CD44 and its isoforms V5 and V6 are overexpressed in human gastric carcinoma. Among the numerous extracellular matrix components, hyaluronate, a CD44 ligand, is of increasing interest in relation to its role in cancer cell development and invasion. By using the dynabead separation method, the SC-M1 cell line was separated into V5 and V6 isoform-positive and -negative populations. The V5 and V6 isoform-negative populations exhibited significantly higher hyaluronate binding activity than the corresponding positive cells. The hyaluronate binding activity of V5 and V6-positive cells could be restored by pretreatment with anti-CD44 V5 and V6 monoclonal antibodies (MAbs). In addition, transfection of aVV5 and V6-negative cells decreased their hyaluronate binding activity to the levels of CD44 V5 and V6-positive cells. Cells transfected with V5 and V6 recovered their hyaluronate binding activity after pretreatment with MAbs against V5 and V6. These data suggest that cell adhesion involving hyaluronate can be regulated by multiple mechanisms, one of which involves alternative splicing of CD44 isoforms.

Evidence of transmission of Mycobacterium tuberculosis by random amplified polymorphic DNA (RAPD) fingerprinting in Taipei City, Taiwan.

AIMS: To determine, by strain identification of Mycobacterium tuberculosis, whether transmission has occurred between individuals or whether new strains are present. METHODS: A rapid protocol for random amplified polymorphic DNA (RAPD) analysis was developed. This protocol was applied to 64 strains of M tuberculosis that had been confirmed by culture and microbiological methods. RESULTS: There are five groups of M tuberculosis prevalent in Taipei city, Taiwan. The major types are groups I and III. Groups I and II had been prevalent until the end of last year when, according to our group analysis, they had been eradicated. However, group III was continuously present from the middle of 1995 to the middle of 1996, and group IV was present at the end of both years, which indicated that both groups were transmitted continuously. These clustered strains had demographic characteristics consistent with a finding of transmission tuberculosis. Also, there were 13 of 64 strains with unique RAPD fingerprints that were inferred to be due primarily to the reactivation of infection. In the drug resistance analysis, the major type represented included group III and part of group IV. CONCLUSIONS: Our preliminary data imply, not only that the prevalence of M tuberculosis in Taipei city is due to transmission rather than reactivation, but that drug resistance also may play a role in tuberculosis transmission.

Apoptosis occurs more frequently in intraductal carcinoma than in infiltrating duct carcinoma of human breast cancer and correlates with altered p53 expression: detected by terminal-deoxynucleotidyl-transferase-mediated dUTP-FITC nick end labelling (TUNEL).

AIMS: We examined the relationship between apoptosis and three different major stages of human breast carcinoma: intraductal carcinoma (DCIS), infiltrating duct carcinoma (IDC) and metastatic carcinoma in lymph nodes. We also determined the correlation between apoptosis and oestrogen receptor (ER), progesterone receptor (PR) and p53. METHODS AND RESULTS: The study investigates the extent of apoptosis in 63 breast carcinomas by in-situ end-labelling, in formalin-fixed, paraffin-processed tissue sections. The 63 breast carcinomas, included 22 DCISs, 26 IDCs, three infiltrating lobular carcinomas (ILC) and 12 metastatic lymph nodes. The apoptotic labelling index was higher in DCIS than IDC and metastatic carcinoma (P < 0.001, P < 0.007, respectively). By immunohistochemistry, we also analysed p53, ER and PR. Apoptosis correlated significantly with p53 (r = 0.748, P = 0.0004) in IDC. Also, ER correlated significantly with PR (r = 0.629, P = 0.00001). No apparent correlation was found between the apoptosis and ER or PR. CONCLUSION: Our data suggest that not only does apoptosis differ between intraductal carcinoma and infiltrating carcinoma but also it might be regulated by altered p53 expression.

Postextubation laryngeal edema in adults. Risk factor evaluation and prevention by hydrocortisone.

OBJECTIVE: To evaluate the risk factors for postextubation laryngeal stridor and its prevention by hydrocortisone in adult patients. DESIGN: Prospective, randomized, double-blind, placebo controlled study. SETTING: Medical and surgical ICU of a tertiary teaching hospital. PATIENTS: 77 consecutive patients of both sexes, who had undergone tracheal intubation for more than 24 h and fulfilled the weaning criteria, were eligible for the study. Patients were excluded if they were less than 15 years of age, had a disease or the surgery of the throat, or had been extubated during the current hospitalization. INTERVENTION: The control group received placebo (normal saline 3 cc) and the experimental group received hydrocortisone 100 mg by intravenous infusion 60 min before extubation. MAIN OUTCOME MEASURES: Patients were observed 24 h after extubation for symptoms or signs of laryngeal edema or stridor: prolonged inspiration with accessory usage of respiratory muscles or crowing sound with inspiration or reintubation. RESULTS: The overall incidence of postextubation stridor was 22% (17/77). Only one patient (1%), who belonged to the control group, needed reintubation. 39% of female patients and 17% of male patients developed stridor. The relative risk of females developing this complication was 2.29. 7/39 of the hydrocortisone group and 10/38 of patients in the control group developed postextubation stridor. CONCLUSIONS: Hydrocortisone did not significantly reduce the incidence of postextubation laryngeal edema or stridor. From the risk factors evaluated, we were unable to demonstrate a statistical correlation between postextubation stidor and the duration of the intubation, the patient's age, the internal diameter of the endotracheal tube, or the route of intubation. However, female patients were more likely to develop this complication.

Down regulation of bcl-2 by p53 in nasopharyngeal carcinoma and lack of detection of its specific t(14;18) chromosomal translocation in fixed tissues.

High levels of bcl-2 protein have been found in a wide variety of human cancers. Since p53 gene inactivation occurs in over half of human cancers, it is possible that loss of p53-mediated repression of bcl-2 gene expression accounts, at least in part, for the frequent abnormalities in bcl-2 protein production seen in tumours. By using immunohistochemical methods, we have analysed thirty-three nasopharyngeal carcinomas for p53 and bcl-2 expression. We found an inverse correlation between the expression of these two proteins (P < 0.001). Moreover, we utilized universal oligonucleotide primers of a region 5' to the bcl-2 MBR and at the 3' end of JH segments to initiate a DNA polymerase chain reaction that amplified these bcl-2-JH junctures. Of the twelve nasopharyngeal carcinomas expressing bcl-2, none showed a t(14;18) chromosome translocation. These findings may indicate potential mechanisms by which bcl-2 regulates apoptosis.

Soluble CD44 isoforms in serum as potential markers of metastatic gastric carcinoma.

A splice variant of CD44 (exon V4-V7) confers metastatic behavior in a rat carcinoma model; aberrant expression of splice variants has been detected on a variety of human tumor cell lines as well as primary and metastatic human tumors, including lymphomas, carcinomas (colon, thyroid, mamma, bladder), and glioma. We used enzyme-linked immunosorbent assay to determine the concentration of soluble CD44 in the serum samples of 10 normal individuals and 41 patients with various stages of gastric cancer. Soluble CD44S and its isoforms, V5 and V6, were present in the serum of normal individuals (288.53 +/- 18.33, 25.49 +/- 1.70, and 148.32 +/- 3.15 ng/ml, respectively). The concentrations of soluble CD44 V5 and V6 were elevated in patients with advanced gastric carcinoma (69.39 +/- 6.06 and 216.62 +/- 32.98 ng/ml, respectively). Serum CD44 V5 concentrations correlated with the extent of tumor invasion (T), the status of lymph node involvement (N), and distant metastasis (M) (TNM staging) (p < 0.05), whereas CD44S did not. These results suggest that detection of abnormal regulation of CD44 splicing could be helpful in gastric cancer diagnosis and disease evaluation.

Evidence that polymorphism of the angiotensin I converting enzyme gene may be related to idiopathic dilated cardiomyopathy in the Chinese population.

Angiotensin I-converting enzyme (ACE) is responsible for the production of angiotension II and the breakdown of kinins, leading to increased blood pressure (BP), induction of vascular smooth muscle cell proliferation, and the stimulation of myocardial-cell hypertrophy. A 287 bp insertion/deletion polymorphism in intron 16 of the ACE gene was examined by polymerase chain reaction in a cross-sectional study of 35 patients with idiopathic dilated cardiomyopathy (IDC) and 35 patients with normally functioning hearts (NT). Compared with the deletion/deletion (D/D) frequency in the control population, the frequency of the deletion allele was 0.757 in IDC patients and 0.600 in NTs; the difference between observed alleles in all subjects in each group was significant (x2 = 3.96; P < 0.05). The data thus provide evidence in favor of an association between idiopathic dilated cardiomyopathy and a polymorphism at the ACE locus (17q23), thus implicating this locus, and possibly a genetic variant of ACE, itself, in human idiopathic dilated cardiomyopathy.

Differential expression of the human metastasis adhesion molecule CD44V in normal and carcinomatous stomach mucosa of Chinese subjects.

BACKGROUND: CD44 is a cell surface adhesion molecule involved in cell-cell and cell-matrix interactions. Tumor cells transfected to overexpress the isoform CD44V readily gain access to lymph nodes and form distant metastases. METHODS: Monoclonal antibodies directed at epitopes common to known CD44 isoforms were used to investigate CD44V expression in 30 normal gastric mucosa tissues, 64 different gastric adenocarcinomas, 20 metastatic adenocarcinoma lymph nodes and 4 established gastric carcinoma cell lines. In addition, CD44V gene expression in six gastric adenocarcinoma tissues and four gastric cancer cell lines were investigated by northern blotting. RESULTS: Immunohistochemistry screening of 30 subjects with normal gastric mucosa did not reveal expression of CD44 variants. Areas of intestinal metaplasia, a precancerous lesion, were stained with antibodies against either V5- or V6-containing isoforms of CD44. Tubular and signet-ring cell types of adenocarcinoma were strongly positive for epitopes encoded by CD44 variants containing exons V5 (41/49 and 10/10, respectively). Some tubular type adenocarcinomas (15/49) also expressed CD44 variants containing the V6 epitope. Tumor differentiation was closely related to CD44 V5 expression (P < 0.001). In addition, 18 of 20 gastric adenocarcinomas metastatic to lymph nodes expressed the V5 epitope of CD44 and 4 of 20 expressed the V6 epitope. Analysis of four established gastric adenocarcinoma cell lines revealed that two had moderate to strong expression of exons V5 and V6 of CD44. An antibody directed against CD44 variants containing exons V8 to V10 strongly stained all gastric adenocarcinoma cell lines. Northern blotting demonstrated that all four tumor cell lines and six gastric carcinoma mucosa tissues expressed CD44V. CONCLUSIONS: Generation of CD44 splice variants may be closely linked with gastric carcinoma tumorigenesis and differentiation. In addition, expression of CD44 variants containing exons V5 and V6 may be used as an indicator of evolving gastric cancer.

Epstein-Barr virus-associated gastric adenocarcinoma in Taiwan.

Fifty-five gastric carcinoma tumors from Chinese patients in Taiwan, including 40 tubular type (one lymphoepthelioma-like carcinoma subtype), eight signet ring cell type, one papillary type, and six mucinous type gastric carcinomas, were investigated for the presence of Epstein-Barr virus (EBV) transcripts by in situ hybridization using fluorescein-conjugated EBV oligonucleotides for EBERs (Epstein-Barr virus early RNAs) expression and the polymerase chain reaction for viral DNA. Epstein-Barr virus was detected in six of 55 lesions (11%), a significantly lower proportion than has been observed in a North American series. Epstein-Barr virus involvement was more common among male patients. Epstein-Barr virus DNA and its EBERs were specifically present within gastric carcinoma and adjacent dysplastic cells but were absent in surrounding lymphocytes and normal gastric mucosa. Epstein-Barr virus DNA and EBERs were found in one sample of lymphoepithelioma-like carcinoma (LELC) and five specimens of typical gastric adenocarcinoma. Among the EBV-positive gastric adenocarcinomas, four were tubular type of varied differentiation and one was signet ring cell type. Furthermore, we evaluated the expression of the latent membrane protein (LMP) with monoclonal antibodies. We found that LMP was expressed in two EBV-positive samples. In addition, the presence of the EBV receptor was studied by probing samples with CD21 monoclonal antibody. Epstein-Barr virus receptor was not detected in any sample. Southern blot analysis indicated single clonal proliferation of tumor cells. These findings confirm and extend the results of Shibata et al. They also indicate that EBV infection might be related to oncogenesis not only in rare gastric cancers that resemble nasopharyngeal lymphoepithelioma but also in typical gastric adenocarcinoma.

Gastric lymphoepithelioma-like carcinoma associated with Epstein-Barr virus proved by in situ hybridization study: a case report.

Undifferentiated gastric carcinoma with prominent lymphoid infiltration and a striking resemblance to nasopharyngeal lymphoepithelioma has rarely been reported. Recently, the Epstein-Barr virus (EBV) genome has been demonstrated in some cases of gastric carcinoma with a morphology similar to undifferentiated nasopharyngeal carcinoma (NPC). One such case has been identified here by immunohistochemistry; it was explored for the EBV genome by RNA in situ hybridization (ISH) with an EBERs (Epstein-Barr virus encoded RNAs) probe. EBV RNAs were detected uniformly in the carcinoma cells or adjacent dysplasia epithelium, but were not present in the related lymphoid infiltration or in normal gastric mucosa. These findings add to the growing body of evidence suggesting the strong association of EBV with gastric lympho-epithelioma-like carcinoma (LELC), and also indicate that a latent EBV infection may play a role in undifferentiated epithelial cells.

Selective expression of CD44 messenger RNA splice variants in four high grade human brain tumour cell lines.

Changes in CD44 transcripts have been previously found to be associated with metastasis in animal models. The purpose of this study was to investigate CD44V changes in four well established high grade human brain tumor cell lines, known to possess prominent invasive behavior. In Northern blot analysis, CD44S and CD44V were expressed strongly in three high-grade glioblastoma multiforme cell lines (GBM 8401, GBM 8909, GBM 8804) and one malignant meningioma cell line (IOMM). By RT-PCR and blot hybridization, three variant transcripts (650 bps, 850 bps, and 1,000 bps) were detected in GBM 8804 and two isoform transcripts (650 bps, 850 bps) were recognized in GBM 8401 and 8909. Further, in a malignant meningioma cell line (IOMM), only one weak isoform (650 bps) was detected. However, by Northern blot analysis, neither CD44S or CD44V could be expressed in normal brain and meningeal tissue. These results indicate that discrete CD44 mRNA splice variants are expressed in high grade glial cell tumors and malignant meningioma and suggest a possible role in the invasion of malignant brain tumors.

The variant mRNA isoform of human metastasis gene (CD44V) detected in the cell lines of human hepatocellular carcinoma.

The interaction of the cell surface receptor CD44 molecular with its ligands (addressin, extracellular matrix etc.,) plays an important role in fulfilling the lymphocyte homing and immune reaction. Recently alternatively spliced products of CD44 gene are found to be involved in tumor metastasis as well. Our report found that CD44 prototype RNA (CD44S) was present in all five tumor cell lines. Isoform CD44 RNA (CD44V) was recognized in three metastasized hepatocellular carcinoma cell lines, J5, HCC36, HEP3B. In addition, the J5 CD44 RNA isoform expressed two distinct transcripts which are of the same size as MDA-231 breast tumor cell line. The MDA-231 CD44 RNA variant (CD44V) has been confirmed to contain metastasis domain 4 and 5. It is implicated that the alternative RNA splicing may also play a major role in hepatocellular carcinoma metastasis.

[Efficacy of disodium cromolyn in adult chronic asthma].

Disodium cromolyn is effective in preventing asthma, but its steroid sparing effect remains controversial. In this prospective cross-over study, we used two different brands of disodium cromolyn, Intal and Ticromil, each for 4 weeks on 7 adult patients with chronic asthma. The dosage was 20 mg qid via a special inhaler. The whole course of the study was 12 weeks for each patient, including 2 run-in weeks and 2 wash-out weeks. Patients were followed up by the same physician at outpatient clinics every 2 weeks to measure FEV1, FVC, peak expiratory flow rate (PEFR), and reversibility after bronchodilator. The physician also assessed the severity of the patients' symptoms via a special score system. Patients recorded symptom severity and frequency of medications at home on daily diary cards. Patients also measured PEFR twice a day. At the end of the treatment, FEV1 was improved more in Intal group; daily prednisolone dosage was markedly reduced in Ticromil group; PEFR was improved in both group; but neither symptom severity nor airway reversibility was significantly changed. When two groups were calculated as a whole, PEFR increased significantly from 294 +/- 20 l/min to 342 +/- 19 l/min (p = 0.0225). The daily prednisolone dosage reduced from 5.87 +/- 0.91 mg to 3.91 +/- 0.60 mg in the 3rd week and to 4.05 +/- 0.63 mg in the 4th week. The differences between these 2 dosages and the baseline dosage were statistically significant (both p < 0.05). Side effects were minimal in all patients. We concluded that after the use of disodium cromolyn for 4 weeks, PEFR was markedly improved and daily prednisolone dosage was also significantly reduced.