Elevation of transforming growth factor alpha and its relationship to the epidermal growth factor and alpha-fetoprotein levels in patients with hepatocellular carcinoma.

A radioimmunoassay for transforming growth factor alpha (TGF-alpha) using synthetic rat sarcoma transforming growth factor and its rabbit polyclonal antibody has been developed. Using radioimmunoassays, the urinary TGF-alpha and epidermal growth factor (EGF) concentrations in 31 patients with hepatocellular carcinoma (HCC), 15 probable HCC, four metastatic liver cancer, and 33 age, sex-matched healthy controls were determined. For the first time, we have shown that the average TGF-alpha concentration for HCC patients was 21.5 +/- 20.3 micrograms per g creatinine, significantly higher than that of healthy subjects, 4.9 +/- 2.8 micrograms per g creatinine (P less than 0.001). There was no statistical difference in the level of EGF between HCC patients and controls (40.9 +/- 29.3 versus 46.2 +/- 16.6 micrograms per g creatinine; P greater than 0.05). The ratio of EGF/TGF-alpha between HCC patients (3.37 +/- 4.42) and controls (15.5 +/- 13.0) was significantly different (P less than 0.001). Among patients, 65% (20 of 31) of HCC cases and 87% (13 of 15) of probable HCC cases showed a marked elevation of TGF-alpha levels. We found only 16% (five of 31) of HCC cases with increased EGF level. EGF excretion was inversely age related. Serum total protein concentration and alkaline phosphatase activity were positively correlated to EGF concentration (r = 0.522, P less than 0.01 and rt = 0.393, P less than 0.05, respectively). There was no correlation between biochemical functions of liver and TGF-alpha concentration in HCC patients. Our results also suggested that TGF-alpha may be a useful complementary tumor marker for management of patients with clinical manifestation of HCC who have low alpha-fetoprotein levels.

Increased IgM-containing circulating immune complexes in patients co-infected with hepatitis C and hepatitis B.

To assess the role of circulating immune complexes (CIC) in chronic hepatitis C virus (HCV) infection, the relative frequency of CIC was determined in 60 patients with chronic hepatitis C alone, 19 patients co-infected with hepatitis B and C, 15 asymptomatic HCV carriers, and 54 healthy controls. Levels of CIC were determined with immunoglobulin-specific C1q-binding and conglutinin (K)-binding immune complex assays. Although there was no statistical difference in the levels of each type of CIC between asymptomatic HCV carriers and healthy controls, elevated levels of CIC containing IgM and IgG were common in patients with chronic HCV infection. Compared to patients with hepatitis C alone, patients co-infected with hepatitis B and C have a higher frequency of abnormal IgM-containing CIC and significantly higher levels of IgM-containing CIC. CIC levels in patients with chronic active hepatitis were significantly higher than those in patients with chronic lobular hepatitis or chronic persistent hepatitis. In conclusion, although CIC containing IgM and IgG were common in patients with chronic hepatitis C, abnormal IgM-containing CIC are the major types of CIC in patients co-infected with hepatitis B and C. An immune-mediated mechanism may play a role in the pathogenesis of chronic hepatitis C.

Clinical relevance of transforming growth factor-beta 1 in the urine of patients with hepatocellular carcinoma.

To assess the clinical relevance of transforming growth factor-beta 1 (TGF-beta 1) in the urine of patients with hepatocellular carcinoma (HCC), TGF-beta 1 was measured, by radioimmunoassay, in 140 patients with HCC, 50 cirrhotic patients, 30 patients with chronic active hepatitis, and 50 healthy controls. The results indicate that there were significantly increased urinary TGF-beta 1 levels in patients with HCC. Raised TGF-beta 1 levels were associated, in a dose-related fashion, with increased risk for development of HCC (odds ratio, 1.05, 95% confidence interval, 1.03-1.07). HCC patients with raised TGF-beta 1 levels had shorter survival than those with normal TGF-beta 1 levels (p = 0.038). TGF-beta 1 levels decreased after successful anticancer therapy (p < 0.0001). There was an inverse correlation between TGF-beta 1 and serum alpha-fetoprotein (AFP) (r = -0.199, p < 0.04). Receiver operating characteristics (ROC) curve analysis indicated that parallel determination of TGF-beta 1 and AFP significantly increased the sensitivity and diagnostic accuracy, with a high specificity. In conclusion, raised urinary TGF-beta 1 was associated with HCC development. It is a predictor of poor prognosis, and a tumor marker for diagnosis and therapeutic follow-up of HCC.

GB virus C/hepatitis G virus infection in chronic hepatitis C patients with and without interferon-alpha therapy.

GB virus C/hepatitis G virus (GBV-C/HGV) RNA, detected by polymerase chain reaction, and antibodies to the GBV-C/HGV envelope protein (anti-E2), detected by an enzyme-linked immunosorbent assay, were used to evaluate both the impact of GBV-C/HGV on the coexistent hepatitis C virus (HCV) infection and the course of GBV-C/HGV infection in chronic hepatitis C patients with and without interferon-alpha (IFN-alpha) treatment. Of the 162 chronic hepatitis C patients treated with INF-alpha, 17.9% were GBV-C/HGV RNA-positive and 18.5% anti-E2-positive (total exposure, 35.2%). Neither present nor past GBV-C/HGV infection had impact on the clinical features, HCV virological characteristics and response to IFN-alpha treatment in chronic hepatitis C patients. Among patients with ongoing HCV/GBV-C/HGV coinfection, 20.7% (6/29) in IFN-alpha-treated patients lost GBV-C/HGV RNA concomitant with anti-E2 seropositivity, which was significantly higher than 4.8% (2/42) in patients without INF-alpha treatment (P<0.05). Based on multivariate analyses, the significant factors associated with clearance of GBV-C/HGV viremia combined with anti-E2 seropositivity were baseline anti-E2 seropositivity and IFN-alpha treatment. In summary, GBV-C/HGV did not alter the course of coexistent HCV. IFN-alpha treatment was effective in some patients against GBV-C/HGV and might facilitate anti-E2 seroconversion in chronic hepatitis C patients with GBV-C/HGV viremia.

Clinical evaluation of the COBAS Amplicor HBV monitor test for measuring serum HBV DNA and comparison with the Quantiplex branched DNA signal amplification assay in Taiwan.

AIMS: To evaluate the performance characteristics and clinical usefulness of the COBAS Amplicor HBV monitor (COBAS-AM) test in Taiwan and to examine its correlation with the Quantiplex branched DNA signal amplification (bDNA) assay for measuring serum hepatitis B virus (HBV) DNA concentrations. METHODS: HBV DNA was measured by the COBAS-AM test in 149 sera from chronic HBV infected patients that had previously been analysed by the bDNA assay. RESULTS: The COBAS-AM test showed good reproducibility, with acceptable intra-assay and interassay coefficients of variation (1.6% and 0.9%, respectively) and good linearity (r2=0.98). The overall sensitivity of the COBAS-AM test was significantly higher than that of the bDNA assay (95.3% v 83.2%): 69.6% of samples with HBV DNA below the detection limit of the bDNA assay could be measured by the COBAS-AM test. There was a significant correlation between the results of the two assays (r=0.901; p<0.0001). On average, the results derived from the COBAS-AM test were 0.55 log lower than those of the bDNA assay. HBV DNA concentrations were significantly higher among HBV e antigen (HBeAg) positive patients than negative ones, and higher among patients with abnormal alanine aminotransferase (ALT) concentrations than those with normal ALT concentrations (p=0.0003). CONCLUSIONS: The COBAS-AM assay, more sensitive in HBeAg negative samples than the bDNA assay, can effectively measure HBV DNA concentrations in Taiwanese patients. HBV DNA values measured by the COBAS-AM test and bDNA assay correlate significantly.

Urinary epidermal growth factor receptor-binding growth factors in patients with cancers of the digestive tract.

To investigate whether the urinary excretion of epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) is increased in patients with cancer of the digestive tract, EGF and TGF-alpha were determined in 109 cancer patients and 40 healthy controls. Excluding EGF in hepatocellular carcinoma (HCC) and TGF-alpha in pancreatic cancer, both growth factors in each cancer group were significantly higher than in the control group. A receiver operating characteristic curve and likelihood ratio were applied to obtain the best diagnostic efficiency. Both EGF and TGF-alpha had high specificity (100%) in all cancer group. The high sensitivity of EGF in gastric cancer (100%) and metastatic liver cancer (93.3%), moderate sensitivity of TGF-alpha in metastatic liver cancer (86.6%), colon cancer (80.0%), and HCC (61.7%) suggested that they might be helpful in identifying these cancers. In conclusion, urinary excretion of EGF and TGF-alpha increased in most cancers of the digestive tract. They may be used as tumor markers.

Epidermal growth factor receptor-binding growth factors in the urine of patients with cancers of the digestive tract.

We aimed to assess the diagnostic application of urinary epidermal growth factor receptor (EGFR)-binding growth factors in cancers of the digestive tract. By radioreceptor assay and radioimmunoassay, we determined these growth factors in 115 patients with various cancers of the digestive tract, 30 patients with benign disease, and 40 healthy controls. The receiver operating characteristic (ROC) curve and likelihood ratio were employed to determine the best diagnostic efficiency. Urinary EGFR-binding growth factors in each cancer group were significantly higher than those in the non-cancer groups. Multivariate analysis indicated that the growth factors, determined by both the radioreceptor assay (odds ratio, 1.184; 95% confidence interval,1.077-1.302; P = 0.001) and radioimmunoassay (odds ratio,1.055; 95% confidence interval, 1.002-1.111; P = 0.039), were associated, in a dose-related fashion, with the presence of cancers. By ROC curve analysis, the optimal cutoff values for EGFR-binding growth factors were 25.5 microg/g creatinine (radioreceptor assay) and 33.6 microg/g creatinine (radioimmunoassay). The resulting sensitivity, specificity, diagnostic accuracy, and positive and negative likelihood ratios were 84.4%. 87.5%, 85.2%, 6.75 and 0.18 (for radioreceptor assay) and 86.1%, 67.5%, 81.3%, 2.64 and 0.21 (for radioimmunoassay), respectively. Except for pancreatic cancer the growth factors showed moderate diagnostic efficiency for the other digestive tract cancers. In conclusion, urinary EGFR-binding growth factors were increased in cancers of the digestive tract. They may be used as diagnostic tumor markers.

Sex differences in relation to serum hepatitis B e antigen and alanine aminotransferase levels among asymptomatic hepatitis B surface antigen carriers.

This study aimed to investigate sex differences in relation to hepatitis B e antigen (HBeAg) and serum alanine aminotransferase (ALT) levels in chronic asymptomatic hepatitis B virus (HBV) infection. HBeAg and ALT level were determined in 636 asymptomatic hepatitis B surface antigen carriers. There was no significant sex differences in the age-adjusted prevalence of HBeAg. Abnormal ALT level (>45 IU/l) was more frequent in carriers with HBeAg (17.5% vs 7.6%; P = 0.001). Multivariate analysis indicated that male sex (odds ratio, 2.0; 95% confidence interval, 1.1-3.6) and HBeAg (odds ratio, 2.6; 95% confidence interval, 1.6-4.3) were independent risk factors for abnormal ALT levels. Male sex and HBeAg-positivity are independent risk factors for abnormal ALT activity in chronic HBV infection. This observation may be related to sex differences in chronic HBV infection.

The serological and molecular epidemiology of GB virus C/hepatitis G virus infection in a hepatitis C and B endemic area.

OBJECTIVES: To investigate the serological and molecular characteristics of GB virus C/hepatitis G virus (GBV-C/HGV) infection in the hepatitis C virus (HCV)/hepatitis B virus (HBV)-endemic areas in Taiwan. METHODS: Sera from 200 residents from Masago, an HCV/HBV-endemic community in Taiwan, and 400 blood donors were tested for GBV-C/HGV RNA by using nested reverse transcription-polymerase chain reaction and for antibodies to GBV-C/HGV E2-protein (anti-E2) by an enzyme-linked immunosorbent assay. Phylogenetic analysis of GBV-C/HGV was performed. RESULTS: The prevalence of GBV-C/HGV viraemia, anti-E2 and GBV-C/HGV exposure among residents of Masago was significantly higher than that among donors (17.0%, 25.5% and 39.5% vs. 3.3%, 7.5% and 10.3%, respectively; all P < 0.0001). In Masago, the prevalence of GBV-C/HGV exposure was significantly higher in residents exposed to HCV than in those without HCV exposure (45.8% vs. 24.1%;P< 0.005). Based on multivariate analyses, HCV viraemia was the only significant factor associated with elevated levels of alanine aminotransferase in Masago. Phylogenetic analysis showed all 34 GBV-C/HGV isolates from Masago clustered within genotype 3. CONCLUSIONS: GBV-C/HGV was highly prevalent in Masago, an HCV/HBV-endemic community in Taiwan. HCV viraemia played the most important clinical hepatopathic role in the area. Infections with other hepatitis viruses did not influence the anti-E2 seroconversion from GBV-C/HGV infections.

The molecular epidemiology and clinical significance of TT virus (TTV) infection in healthy blood donors from southern Taiwan.

To evaluate the molecular epidemiology and clinical significance of th

Duplex pulsed Doppler sonography in the differential diagnosis of hepatocellular carcinoma and other common hepatic tumours.

180 previously untreated consecutive patients with liver tumours (308 lesions), including 104 hepatocellular carcinomas (148 lesions), 43 metastases (116 lesions) and 33 haemangiomas (44 lesions), were studied to determine the value of duplex sonography in the differentiation of hepatocellular carcinoma from other tumours. For lesions measuring less than or equal to 5 cm in diameter, hepatocellular carcinoma demonstrated the highest rate and haemangioma demonstrated the lowest rate of Doppler signals from within the lesions. To differentiate malignancy from haemangioma, the presence or absence of Doppler signals from these lesions were used as criteria. The specificity and positive predictive value were very high (100%, 100%), but the sensitivity, negative predictive value and accuracy were low (61.5%, 48.3%, 71.7%, respectively). With one exception, all lesions measuring less than 3 cm in diameter with detectable Doppler signals were hepatocellular carcinoma. Using these results it is possible to differentiate hepatocellular carcinoma from metastases and haemangioma with high sensitivity, specificity, positive and negative predictive value, and accuracy (80.8%, 96.4%, 95.5%, 84.4%, 88.9%, respectively, for metastases; 80.8%, 100%. 100%, 81.5%, 89.6%, respectively, for haemangioma). We conclude that Doppler signals from within a lesion in combination with its size can aid differentiation of hepatocellular carcinoma from two other kinds of common hepatic tumour.

Percutaneous ethanol injection of the supplying artery to hepatocellular carcinoma that is not amenable to conventional treatment.

The purpose of this study was to evaluate the clinical usefulness of ultrasound-guided percutaneous ethanol injection of the supplying artery (PEISA) to the tumour in the palliative management of hepatocellular carcinoma (HCC) that is not amenable to conventional treatments. A total of 23 cases of HCC, measuring from 3.1 cm to larger than 15 cm (median 5.4 cm) in 17 cirrhotic patients, were treated by PEISA. PEISA was used to control rapid growth of the tumour in seven patients and to reduce abdominal discomfort caused by rapid expansion of the tumour in 10 patients. Tumours with arterial Doppler signals persisting after PEISA underwent repeated treatment. The follow-up period ranged from 2-48 months. PEISA was achieved in 69 out of 76 attempts (90.8%). The amount of ethanol injected on each occasion ranged from 2.5-33 ml. Follow-up colour Doppler scanning showed complete elimination of tumour Doppler signals in 22 out of 23 lesions (95.7%). Following treatment, one tumour disappeared, 13 tumours shrank and nine tumours were unchanged in size. All patients with abdominal discomfort had relief after treatment. The common complications of PEISA were local pain and fever. In conclusion, PEISA is effective at treating painful HCC unsuitable for conventional treatment.

Treatment of hepatocellular carcinoma smaller than 5 cm by transcatheter arterial chemoembolization.

To assess the effectiveness of transcatheter arterial chemoembolization (TACE) in the treatment of small hepatocellular carcinoma (HCC) and to analyze the prognostic factors, a total of 77 patients with histologically proven HCC, < or = 5 cm in diameter, were enrolled for the study and followed for more than 2 years, The overall cumulative 1-, 2-, and 3-year survival rates were 79.2%, 50.6% and 36.7%, respectively. The median survival time of Child-Pugh's A patients was 990 +/- 146 days, which was significantly better than the median survival time of Child-Pugh's B patients (450 +/- 82 days) Furthermore, positive HBeAg and alpha-Fetoprotein (AFP) were factors close to statistically significant. In contrast, sex, age, HBsAg, Anti-HCV, tumor type, tumor size and tumor number were not related to the prognosis in small HCC who received TACE.

The epidemiology of TT virus (TTV) infection in a hepatitis C and B virus hyperendemic area of southern Taiwan.

TT virus (TTV) is a newly isolated DNA virus from the serum of a patient with posttransfusion hepatitis of unknown etiology in 1997. To evaluate the clinical and molecular characteristics of TT virus (TTV) in a hepatitis C virus (HCV) and B (HBV) hyperendemic area (Masago), 200 residents were enrolled in the study. The sera were tested for alanine aminotransferase (ALT), HCV RNA and GB virus C/Hepatitis G virus (HGV) RNA, TTV DNA, HBsAg, anti-HCV and antibodies to HGV E2-protein (anti-E2). TTV DNA was positive in 99 of the 200 sera with a prevalence rate of 49.5%. The prevalence of HBsAg, anti-HCV, HCV RNA, HGV RNA, anti-E2 and HGV exposure (defined as positive for serum HGV RNA and/or anti-E2) was 38.9%, 69.5%, 64.5%, 17.0%, 25.5% and 39.5%, respectively. Neither clinical nor virological factors were associated with TTV viremia. The rate of ALT abnormality was significantly elevated in HCV RNA-positive (34.9%) than -negative (7.0%) residents (p < 0.001). HCV viremia was the only factor significantly associated with ALT elevation by multiple logistic regression (odds ratio: 6.96; 95% C.I.: 2.60-18.7). We concluded that in this HCV/HBV hyperendemic area, the prevalence of TTV DNA was high. No significant clinical factor was observed to be associated with TTV infection. TTV infection is not related to abnormal ALT levels and ALT abnormality was mainly attributable to HCV but not TTV, HBV or HGV infection.

Bioinspired artificial photonic nanoarchitecture using the elytron of the beetle Trigonophorus rothschildi varians as a 'blueprint'.

An unusual, intercalated photonic nanoarchitecture was discovered in the elytra of Taiwanese Trigonophorus rothschildi varians beetles. It consists of a multilayer structure intercalated with a random distribution of cylindrical holes normal to the plane of the multilayer. The nanoarchitectures were characterized structurally by scanning electron microscopy and optically by normal incidence, integrated and goniometric reflectance measurements. They exhibit an unsaturated specular and saturated non-specular component of the reflected light. Bioinspired, artificial nanoarchitectures of similar structure and with similar properties were realized by drilling holes of submicron size in a multilayer structure, showing that such photonic nanoarchitectures of biological origin may constitute valuable blueprints for artificial photonic materials.

Taiwan's population and family planning efforts: an historical perspective.

The unprecedented decline of Taiwan's natural population increase rate from 3% in 1963 to 1.9% in 1973, attributed primarily to an increasing use of contraception, brought universal attention to one of the first and most successful national family planning programs, at a time when successes were few. Taiwan met its goal of decreasing its natural increase rate despite its young population, increasing numbers of women of childbearing ages, a strong preference for sons and a per capita income at the outset in 1963 of less than U.S.$200. Recognition of this achievement brought thousands of professionals, particularly from developing countries, to study the program firsthand: more than 3,000 during 1970--73 alone. This was matched by an avalanche of publications about the program that appeared around the world, written by practitioners, academics and others. This article tells the story of this success.

Serum insulin-like growth factor-II as a serologic marker of small hepatocellular carcinoma.

OBJECTIVE: Alpha-fetoprotein (AFP) is not a useful tumor marker for diagnosis of small hepatocellular carcinoma (HCC). There is over-expression of insulin-like growth factor (IGF)-II in HCC tissue. This study investigates the diagnostic application of IGF-II in small HCC. MATERIAL AND METHODS: Serum levels of IGF-II and AFP were determined in 41 patients with small cirrhotic HCC (< or = 3 cm), 41 sex- and age-matched patients with cirrhosis alone (LC), and 41 healthy adults. The optimal cut-off values for diagnosing HCC were determined with receiver operating characteristics (ROC) curve. RESULTS: Both IGF-II and AFP levels in HCC were higher than those in LC patients or controls (each p = 0.0001). The IGF-II levels in LC patients were lower than those in controls (p = 0.001). In HCC patients, multivariate analysis indicated that that both IGF-II (odds ratio, 4.54; 95% confidence interval, 2.15-9.55; p = 0.0001) and AFP (odds ratio, 1.05; 95% confidence interval, 1.01-1.08; p = 0.003) were found to be associated with an increased risk of presence of HCC. The optimal cut-off values of IGF-II (4.1 mg/g prealbumin) and AFP (50 ng/ml) were determined with ROC curves. The sensitivity, specificity, and diagnostic accuracy values for IGF-II were 63%, 90%, and 70%, respectively. Those for AFP were 44%, 95%, and 70%, respectively. Determination of both markers in parallel significantly increase the diagnostic accuracy (88%) and sensitivity (80%), with a high specificity (90%). CONCLUSIONS: Serum IGF-II level can be used as an independent serologic marker or a complementary tumor marker to AFP for diagnosis of small HCC.

Hepatitis C virus antibody in hepatocellular carcinoma in Taiwan.

The prevalence of antibody to hepatitis C virus (anti-HCV) was investigated in patients with hepatocellular carcinoma (HCC), and correlated with the clinical features. Anti-HCV was detected in 129 histology or aspiration cytology proven HCC patients and 54 healthy controls. Anti-HCV was examined by the HCV EIA (Abbott Laboratories). All healthy controls were anti-HCV-negative. Nineteen of 81 (23.5%) hepatitis B surface antigen (HBsAg)-positive HCC patients were positive for anti-HCV. Anti-HCV was found among 60.4% (29/48) of HCC patients without detectable HB-sAg. Forty-eight of 129 (37.2%) HCC patients were positive for anti-HCV. There was a significant difference in the prevalence of anti-HCV between patients with HBsAg (23.5%) and those without HBsAg (60.4%, P = 0.0001). However, irrespective of the status of HBsAg, there was no statistical difference in sex, age, routine liver function tests, alpha-fetoprotein concentration, or associated cirrhosis between patients with anti-HCV and those without. The results imply that hepatitis C virus may play a role in the pathogenesis of HCC.

Microbiological analysis of 5-formyltetrahydrofolic acid and other folates using an automatic 96-well plate reader.

The growth of auxotrophic bacteria remains the method of choice for the determination of biologically active folate metabolites in plasma. This report describes a microbiological assay for folates adapted to use disposable 96-well plates and an automatic plate reader. The modifications in the assay decreased reagent costs and made the analysis of hundreds of samples per day possible with a sensitivity limit of 10 fmol of (6S)-5-formyltetrahydrofolic acid. This limit compares favorably with that of previously reported, more laborious methods. The unnatural 6R diastereomer of 5-formyltetrahydrofolic acid did not interfere with the microbiological assay of the natural 6S diastereomer.

Isolation and characterization of Streptomyces lividans mutants deficient in intraplasmid recombination.

Plasmid pIF132 containing two direct repeats of the mel (melanin) sequence was used to monitor intraplasmid recombination. Five mutants of Streptomyces lividans TK64 deficient in intraplasmid recombination were isolated. Four contained additional defects in aerial mycelium formation, pigmentation, and nutrient requirements; among these two showed extensive amplification of chromosomal sequences. Mutant JT46 had no pleiotropic defects but had the most severe blockage in recombination. Only one of the mutants was slightly more sensitive to UV and two were slightly more sensitive to mitomycin C. Plasmid pWCL1 (containing pIJ702, pUC12, and HBVsAg sequences; Lee et al. 1986) could not stably replicate in TK64 without spontaneous deletions. In contrast the mutant JT46 maintained the integrity of pWCL1 much more stably.