HIGH RATE OF HEPATITIS B VIRUS MOTHER-TO-CHILD TRANSMISSION IN LAO PEOPLE'S DEMOCRATIC REPUBLIC.

Hepatitis B virus infection is endemic in Lao People's Democratic Republic (PDR). Among 3,000 pregnant women attending an antenatal clinic at Mother and Child Hospital in Vientiane, Lao PDR, 5.8% were HBsAg positive by a rapid test. Among serum samples of 47 infants aged 9-12 months born to HBsAg-positive mothers, 38% were anti-HBs negative. Percent anti-HBs negative children is significantly higher in those born to HBeAg positive mothers than in those born to HBeAg negative mothers (60% vs 25%, p < 0.05). Out of 47 HBsAg-positive mothers, 10 had infants who were HBsAg positive. None of the infants born to HBsAg negative mothers became HBsAg positive but 10/19 (52.6%) of infants born to HBeAg positive mothers became HBsAg positive. This high rate of mother-to-child transmission of HBV in an endemic country is of concern and indicates that routine vaccination program for Lao infants needs strengthening.

Significance of Hepatic Insulin Clearance in Patients with Chronic Hepatitis C and Non-alcoholic Fatty Liver Disease.

Objective Hyperinsulinemia plays an important role in the pathophysiological processes of chronic hepatitis C (CHC) and non-alcoholic fatty liver disease (NAFLD). However, there are few reports on hepatic insulin clearance in patients with these diseases. Methods A total of 74 CHC patients and 37 NAFLD patients were enrolled in this study. We evaluated their hepatic insulin clearance, insulin sensitivity and ß-cell function with an oral glucose tolerance test. Results Hepatic insulin clearance in the patients with CHC was significantly correlated with platelets (r=0.271, p=0.020) and liver fibrosis (r=-0.234, p=0.045) and was significantly affected by both steatosis (mild: 0.157±0.078, severe: 0.114±0.053, p=0.024) and fibrosis (mild: 0.167±0.0857, severe: 0.125±0.052, p=0.010). There were no significant differences in (homeostasis model assessment) HOMA-ß among steatosis and fibrosis stages. In the NAFLD patients, those with severe fibrosis had significantly reduced hepatic insulin clearance (mild: 0.135±0.045, severe: 0.098±0.031, p=0.013) and significantly increased HOMA-ß (mild: 115.6±67.1, severe: 172.8±65.7, p=0.018) compared with the patients with mild fibrosis. Conclusion Liver fibrosis development is associated with hepatic insulin clearance in both the CHC and NAFLD patients.

Human papillomavirus infection induces NF-κB activation in cervical cancer: A comparison with penile cancer.

This study aimed to determine the relationship between human papillomavirus (HPV) and nuclear factor-κB (NF-κB) in cervical cancer using 62 tissues of cervical cancer, and to compare the findings to penile cancer. HPV-DNA integration is a crucial factor for malignant transformation in cervical cancer and can be identified using in situ hybridization. Of the 62 cases, HPV infection was detected in 28 (45.2%). This frequency was lower than in penile cancer (68.2%) as shown by our previous study. The earliest age of onset of cervical and penile cancer was 18 and 35, respectively, whereas the mean age of the initial diagnosis of cervical and penile cancer was 50.1 and 59.6, respectively. The discrepancies of HPV prevalence, earliest ages of onset and mean ages between cervical and penile cancer patients may result from the gender-based synergistic action of HPV associated with multiple epidemiological co-factors. Of the 28 HPV-infected cases, NF-κB expression was observed in the nucleus in 18 (64.3%), in the cytoplasm in 19 (67.9%) and in the nucleus and/or cytoplasm in 27 cases (96.4%). The overexpression of NF-κB in cervical cancer cases suggests that NF-κB activation is a key modulator in driving chronic inflammation to cancer.

Human papillomavirus genotypes in penile cancers from Japanese patients and HPV-induced NF-κB activation.

The causal relationship between chronic inflammation and cancer is widely accepted. Numerous investigations have identified nuclear factor-κB (NF-κB) as an important modulator in driving chronic inflammation to cancer. This study aimed to determine the prevalence of human papillomavirus (HPV) in penile cancer in Japanese patients and whether NF-κB is subsequently overexpressed in penile cancer. Thirty-four specimens of penile tissue (16 malignant and 18 benign cases) were examined to determine the association of HPV infection. An in situ hybridization (ISH) method was used to detect and localize HPV-DNA. A sensitive HPV polymerase chain reaction (PCR) procedure was used for the detection of HPV-DNA, and DNA sequencing was used to identify the HPV genotype. HPV-DNA was detected in 37.5 and 75% of cases of penile cancer, using ISH and PCR, respectively. Our efforts to detect HPV genotypes were unsuccessful as HPV-DNA could not be extracted from these materials. Using ISH, a prevalence of 68.2% of HPV infection was found in penile cancer in Kenyan patients in east Africa. In the present study, all 9 HPV-positive cases, (100%) were NF-κB-positive in the nucleus and/or cytoplasm. In contrast, of the 25 HPV-negative cases, 15 (60%) were NF-κB-positive in the nucleus and/or cytoplasm. Therefore, ISH is a method which is able to prove infection of a large quantity of HPV more effectively when compared with PCR. Thus, a large quantity of HPV infection leads to the activity of NF-κB. The most prevalent genotype was the HPV-22 found in 83.3% of the penile cancer cases. In addition, HPV-11 was found in 81.8% of the non-cancer cases. For cases with a high level of infection, the activity of NF-κB increased compared with those with a low level of HPV infection.

Relationship among human papillomavirus infection, p16(INK4a), p53 and NF-κB activation in penile cancer from northern Thailand.

Human papillomavirus (HPV) E6 and E7 oncoproteins are essential factors for HPV oncogenesis. These E6 and E7 gene products play a central role in the induction of malignant transformation by interacting with several cellular regulatory proteins such as p16(INK4a), p53 and nuclear factor κB (NF-κB). In the present study, conducted in northern Thailand, HPV-DNA was detected in penile cancer cases using an in situ hybridization procedure and p16(INK4a), p53 and NF-κB were detected by immunohistochemistry. Using the cell cycle regulatory proteins p16(INK4a) (61.5%) and p53 (71.8%), it was found that of the 51 cases, 39 (76.5%) were HPV-DNA-positive in penile cancer. On the other hand, 25% p16(INK4a) and 75% p53, respectively, were found in HPV-negative cases. Prevalence of HPV infection (76.5%) was shown in penile cancer cases in northern Thailand. No difference was found between HPV-positive and HPV-negative cases with respect to the presence of the cell cycle regulatory protein p53. On the other hand, p16(INK4a) was found to be different between HPV-positive and HPV-negative cases. Inactivation of tumor suppressor genes, such as p16(INK4a) and p53, to genetic instability, cell immortalization, accumulation of mutations and cancer formation, with or without HPV and irrespective of HPV infection, is therefore suggested. Of the 39 HPV-positive cases, 35 (89.7%) were NF-κB-positive in the nucleus, 29 (74.4%) in the cytoplasm and 37 (94.9%) in the nucleus and/or cytoplasm. NF-κB was detected in 4 (33.3%) of the 12 HPV-negative cases. Therefore, we propose that penile cancer cases with HPV infection are more likely to activate NF-κB than those without HPV infection.

Genotypic distribution of hepatitis C virus in voluntary blood donors of northern Thailand.

The purpose of this study was to determine the prevalence and distribution of HCV genotypes among voluntary blood donors in northern Thailand. From 1998 to 2000, 167 serum samples which tested positive for anti-HCV antibodies in the screening of voluntary blood donors from 5 provinces in northern Thailand were selected for genotyping. Viral RNA was extracted from the sera. The core-E1 region of the HCV-RNA genome was amplified using a OneStep RT-PCR kit. The core-E1 amplicon was sequenced and the HCV genotype was assigned by comparing with the reference sequences available in the GenBank database. Of 167 anti-HCV positive serum samples, 126 (75.4%) contained HCV RNA as detected by PCR. HCV genotype 3 was the most predominant genotype (39.6%), of which 33.3% belonged to genotype 3a and 6.3% to 3b. Genotype 6 was detected in 31%, and genotype 1 was detected in 27.8%. Of the genotype 1 isolates, 14.3% were la, 12.7% were 1b, and 0.8% were 1c. Two HCV isolates detected in the present study were untypeable. About 75% of anti-HCV positive blood donors had chronic HCV infection. In northern Thailand, genotype 3a was the most predominant genotype, while genotype 6, 1a and 1b were also commonly found. The genotypic distribution of HCV isolates from various regions of Thailand were more or less similar. Nevertheless, in this study, the prevalence of HCV genotype 6 (31%) was higher than previously reported by others (8-18%). Phylogenetic analysis of the HCV isolates detected in the present study was also performed.

Synaptophysin immunoreactivity in adrenocortical adenomas: a correlation between synaptophysin and CYP17A1 expression.

DESIGN AND METHODS: The adrenal cortex is not considered to be an intrinsic part of the diffuse neuroendocrine system, but adrenocortical neoplasms possess neuroendocrine properties. In this study, we examined synaptophysin (SYP) and neural cell adhesion molecule (NCAM) expression in adrenocortical adenomas in relation to adrenal function. RESULTS: Immunohistochemical analysis showed that 50.7 and 98.6% of the cortical adenomas showed SYP and NCAM immunoreactivities respectively. There was no apparent difference in NCAM immunoreactivity among the adenomas. However, the immunostaining for SYP was significantly stronger in cortisol-producing adenomas (CPA) than in aldosterone-producing adenomas (APA), nonfunctioning adenomas (NFA), showing no clinical or endocrinological abnormality, or adenomas associated with preclinical Cushing's syndrome (preCS). Western blotting and real-time PCR demonstrated that the expression level of SYP protein and mRNA was significantly higher in CPA than in APA or NFA. Additionally, the SYP mRNA level showed a positive correlation with CYP17A1 mRNA. In addition to the plasma membrane, mitochondria, and smooth endoplasmic reticulum, SYP immunoreactivity was detected in the Golgi area, which is known to be involved in the regulation of mitochondrial cholesterol and the transport of steroid intermediates. It was unexpected that the ratio of positive cells for SYP in preCS was less than that in APA and NFA. However, further examination is required, because the number of preCS cases we investigated was very small. CONCLUSIONS: We propose that SYP expression in adrenocortical cells may be involved in some aspect of adrenal function such as transport or secretion of glucocorticoids.

Possible new hepatitis B virus genotype, southeast Asia.

We conducted a phylogenetic analysis of 19 hepatitis B virus strains from Laos that belonged to 2 subgenotypes of a new genotype I. This emerging new genotype likely developed outside Southeast Asia and is now found in mixed infections and in recombinations with local strains in a geographically confined region.

Human papillomavirus infection in oral verrucous carcinoma: genotyping analysis and inverse correlation with p53 expression.

OBJECTIVE: Verrucous carcinoma (VC) is a rare subtype of squamous cell carcinoma, occurring mostly in oral mucosa. To clarify the role of human papillomavirus (HPV) in VC tumorigenesis, we investigated localization and genotypes of HPV and p53 expression in oral VC. METHODS: We studied paraffin-embedded specimens of 23 VCs and 10 control non-neoplastic lesions in oral mucosa. To investigate HPV infection, HPV genotypes and p53 expression, we respectively employed in situ hybridization (ISH), sequence analysis following short PCR fragment-PCR assay and immunohistochemistry. RESULTS: Of the 23 VC specimens, 11 (48%) had HPV-DNA (detectable by PCR), and 6 (26%) had intranuclear HPV in the upper portion of the squamous epithelium (detectable by ISH). Nine of the 11 PCR-positive specimens showed multiple infections with low- and high-risk HPVs. No HPV-16 infection was detected. Although HPV-6 and HPV-18 were frequently detected by PCR, no HPV could be found in control specimens by ISH. p53 expression was inversely correlated with HPV infection. CONCLUSION: Thus, multiple infections with low- and high-risk HPVs and their rapid replication during hyperkeratinization may participate in the histogenesis of oral VC. Oral VC tumorigenesis may involve the inactivation of p53, which is associated with HPV infection.

Genotypes of hepatitis B virus among children in Chiang Mai, Thailand.

In sub-Saharan Africa, the Pacific, and particularly Asia, hepatitis B virus (HBV) infection is highly endemic, the most common route of transmission is perinatal. To minimize the number of horizontal transmissions, we determined the prevalence of HBV genotypes among children in northern Thailand. From a survey of 1,231 schoolchildren in Chiang Mai during 1998 to 2000, 55 (4.5%) were found positive for HBsAg. Fifty-three HBsAg-positive samples were available for this study. These came from 28 girls (52.8%) and 25 boys (47.2%), age 5-16 years, with a mean age of 12.8 (+/-2.6) years. The laboratory method was based on a multiplex-PCR for the detection of 6 HBV genotypes (A-F). Among 53 HBsAg positive cases, 48 (90.6%) were genotype C, followed by 4 cases of genotype B (7.5%), and 1 case (1.9%) with mixed infection with genotypes B and C. The high prevalence of HBV genotype C follow by genotype B is similar to that found among blood donors in northern Thailand and the nationwide epidemiological survey conducted in 2004. Perinatal transmission may play an important role in the spread of the virus in this area, as in other Asian countries, where genotypes C and B are highly prevalent.

Seroprevalence of hepatitis B and C virus infections among Lao blood donors.

There have been no previous reports of the prevalence of hepatatis B virus (HBV) and hepatitis C virus (HCV) infections in Lao PDR. From 2003 to 2005, 13,897 first-time blood donors were screened for the presence of hepatitis B surface antigen (HBsAg) and hepatitis C virus antibody (anti-HCV). The seroprevalence of HBsAg positive blood donors was 8.7%. The prevalence among males (9.7%) was higher than in females (6.2%). The prevalence of anti-HCV positive blood donors was 1.1%, with no significant differences between males (1.1%) and females (1.0%). Annual positive rates for HBsAg and anti-HCV during the years 2003 to 2005 did not differ significantly. Lao PDR has a high endemicity of HBV carriers (8.7%). Dual infection with HBV and HCV was 0.12%. For preventing HBV infection, the country introduced DPT-Hepatitis B vaccines into the National Immunization Program in 2001. The large reservoir of HBV and HCV infections will cause an enormous burden of patients with cirrhosis and hepatocellular carcinoma in the future.

Ectopic adrenal tissue in the thorax: a case report with in situ hybridization and immunohistochemical studies.

Ectopic or accessory adrenal tissues are usually found in the upper abdomen or along the path of descent of the gonads. The occurrence of supradiaphragmatic adrenal tissue is extremely rare. We report a case of ectopic adrenal tissue composed of both cortical and medullary cells in a 99-year-old woman. The lesion was found incidentally in the paratracheal region at autopsy. We performed in situ hybridization and immunohistochemistry to confirm that the ectopic adrenal tissue possessed the same steroidogenesis as a normal adrenal gland. The ectopic adrenal tissue was encapsulated by fibrous tissue and composed of cells expressing all steroidogenic enzyme mRNAs. The centrally located cells showed immunoreactivities for tyrosine hydroxylase (TH), dopamine beta hydroxylase (DBH), and phenylethanolamine-N-methyltransferase (PNMT). Expression of ACTH receptor (ACTHR) was also evident. These findings indicated that this ectopic adrenal tissue had the capability for steroid and catecholamine biosynthesis under the control of ACTH, and that it might function adequately even under a condition of bilateral adrenal insufficiency.

Role of growth hormone, insulin-like growth factor 1 and insulin-like growth factor-binding protein 3 in development of non-alcoholic fatty liver disease.

BACKGROUND AND AIMS: Pituitary dysfunction including growth hormone (GH) deficiency may be associated with non-alcoholic fatty liver disease (NAFLD). Since the relationships among GH, IGF-1, IGFBP-3, and development of NAFLD without hypopituitarism are unclear, we examined the role of these hormones in the development of NAFLD based on clinical, laboratory and liver histology data. PATIENTS AND METHODS: A total of 55 consecutive patients (20 males and 35 females) with NAFLD. RESULTS: Aspartate amino transferase (AST), AST/ALT, platelet count and IGF-1, levels were significantly associated with differences in fibrosis, since these variables differed between stage 0-1 and stage 2-3 NAFLD. In multivariate analysis, platelet count (P = 0.0223, relative risk (RR), 5.899; 95% confidence interval (CI), 1.288-27.017), and IGF-1 (P = 0.0363, RR, 4.568; 95% CI, 1.101-18.945) showed significant associations with stage 2-3 NAFLD. Additionally, hyaluronic acid levels had a negative relationship with IGF-1 and the IGF-1/IGFBP-3 ratio. There was no relationship of fibrosis with GH level, but decreased GH (P = 0.0414, RR, 0.199; 95% CI, 0.042-0.989) was significantly associated with steatosis of stage 2-3. Low GH/IGF-1 and GH/IGFBP-3 ratios were found in advanced steatosis. CONCLUSION: GH, IGF-1 and IGFBP-3 are associated with hepatic fibrosis and steatosis in NAFLD. Low levels of IGF-1 might be associated with fibrosis while low level of GH with hepatic steatosis.

The prevalence of human papillomavirus genotypes in penile cancers from northern Thailand.

The highest frequency of penile cancer occurs in Asia, Africa, and Latin America, and there have been a few reports concerning the association of penile cancer with human papillomavirus (HPV) infection in these areas. The objective of this study was to determine the relation between penile cancer and the prevalence of HPV genotypes in northern Thailand. Eighty-eight specimens of penile tissue (65 malignant, 1 pre-malignant, and 22 benign cases) were examined to determine the association of HPV infection. An in situ hybridization (ISH) method was used to detect and localize HPV-DNA. Sensitive HPV polymerase chain reaction (PCR) procedure was used for detection of HPV-DNA, and DNA sequencing was used to identify the HPV genotype. HPV-DNA was detected in 53.8% and 81.5% of cases of penile cancer, using ISH and PCR, respectively. The high-risk HPV-16, most commonly associated with penile cancer in previous reports, was found in only one case in this study. The most prevalent genotype was the high-risk HPV-18, found in 55.4% of the cases (32.3% single and 23.1% multiple infection) followed by the low-risk HPV-6, found in 43.1% of the cases (24.6% single and 18.5% multiple infection). In this study, penile cancer was found to be highly correlated with HPV-DNA. Specifically, infection with both the low-risk HPV-6 and the high-risk HPV-18 is the characteristic prevalence of HPV genotypes in penile cancer in this area.

Genotypes of hepatitis B virus among voluntary blood donors in northern Thailand.

There are distinct ethnogeographic variations for the distribution of various hepatitis B virus (HBV) genotypes, and pathogenic and therapeutic differences are also observed. In general, genotype B infection has a relatively better prognosis than genotype C. In Thailand, genotypes C and B were reported as the major genotypes; however, there were no previous reports of HBV genotyping in the north of the country. From 1998 to 2000, 216 HBsAg-positive serum samples (164 males and 52 females, aged 16-52 years), were screened and collected from voluntary blood donors in four provinces of northern Thailand. The method of Naito et al. was employed in this study, with the multiplex-PCR approach and genotype-specific primers to identify genotypes A-F. We found that the HBV genotype C was highly predominant in northern Thailand (89.3%), when compared with the previous reports of genotype C distribution among voluntary blood donors from other areas in the country (50-65%), followed by genotype B (7.4%), mixed infection of genotype B and C (1.9%) and genotype A (0.5%). Four samples (1.9%) were unclassifiable. There was no significant difference of genotype distribution among four northern Thai provinces or each age group.

Hepatitis B-related polyarteritis nodosa presenting necrotizing vasculitis in the hepatobiliary system successfully treated with lamivudine, plasmapheresis and glucocorticoid.

A 64-year-old man was admitted for alithiasic cholecystitis. Necrotizing vasculitis was detected in a gallbladder obtained at the cholecystectomy. Slight elevation of transaminases, HBe antigens and hepatitis B-DNA (HBV-DNA) were detected in the patient. Intrahepatic necrotizing vasculitis was also detected in the liver biopsy specimen, and he also suffered from peripheral neuropathy of suddenly onset. Based on the diagnosis of hepatitis B-related polyarteritis nodosa, lamivudine was initially administered, followed by plasmapheresis and glucocorticoid steroid therapy. These treatments brought satisfactory improvement of polyarteritis nodosa without exacerbation of liver function.

Evaluation of a hepatitis B vaccination program in Chiang Mai, Thailand.

Chiang Mai is a province in northern Thailand that started a vaccination program for hepatitis B virus (HBV) infection in 1989. In this paper, we report the long-term efficacy of this program. Of children aged 4-9 years, 65.7% had a complete course and 3.8% had an incomplete vaccination course. Urban schoolchildren had higher percentage of HB vaccination than rural schoolchildren (89.1% vs 46.9% for the complete course, p < 0.001). The overall prevalence rate of HBsAg in Chiang Mai schoolchildren was 1.2%, with no significant differences between gender (p = 0.496) and school areas (p = 0.477). Anti-HBc antibodies were detected in 6.9% of children. Overall, 26.2% of children had protective levels of anti-HBs antibodies (> 10.0 mlU/ml), and 11.2% had low levels of these antibodies (1.0-9.9 mlU/ml). Compared to previous reports, our results show a lower percentage of anti-HBs antibodies, 33.8% of children age 4 years had protective anti-HBs antibodies, dropping to 18.4% by age 9 years. Among those anti-HBs seropositive, 9.1% were anti-HBc positive, indicating a natural infection with HBV. We found a small number of children, despite adequate immunization, developed HBV infection.

An autopsy case of acute pancreatitis with a high serum IgG4 complicated by amyloidosis and rheumatoid arthritis.

We report an autopsy case of acute pancreatitis with a high serum IgG4 concentration complicated by systemic amyloid A amyloidosis and rheumatoid arthritis (RA). The patient was a 42-year-old Japanese female with a 22-year history of rheumatoid arthritis. She was diagnosed with myasthenia gravis when she was 31-year old. At the onset of pancreatitis, the patient was anti-nuclear antibody-positive, and had high serum gamma globulin and IgG4 levels. Dexamethasone and conventional therapy induced clinical remission and significantly decreased the serum IgG4 and gamma globulin. However, despite the decreased disease parameters, the patient developed a bleeding pseudocyst and died of cardiac failure. In the autopsy examination, it was determined that pancreatitis was probably caused by ischemia due to vascular obstruction caused by amyloid deposition in the pancreas. Even though acute pancreatitis is a rare complication in RA patients, we speculate that an autoimmune pancreatitis-related mechanism and ischemia due to vascular obstruction by amyloid deposition might be attributable to a single source that leads to acute pancreatitis in our particular case.

Metastatic low-grade endometrial stromal sarcoma of the sigmoid colon three years after hysterectomy.

A 49-year-old woman, who had undergone hysterectomy for low-grade endometrial stromal sarcoma (ESS) 3 years ago, presented with a 2-wk history of lower abdominal pain. Barium enema and sigmoidoscopy disclosed a polypoid submucosal tumor. Histopathologic features of biopsy specimens from the lesion were similar to those of the resected uterine ESS. Under the diagnosis of metastatic ESS of the sigmoid colon, sigmoidectomy was performed. Microscopic examination demonstrated dense proliferation of spindle cells with little nuclear atypia, which were sometimes arranged in whorled pattern around abundant arterioles. Mitotic count is below 1 in 10 high-power fields. Immunohistochemically, the neoplastic cells were strongly positive for vimentin, estrogen receptor and progesterone receptor but negative for alpha-smooth muscle actin, S-100 protein and CD34. Thus, a final diagnosis of low-grade ESS metastasis to the sigmoid colon was made. Her postoperative course was uneventful and hormonal therapy with progestational agents is entertained.