HPV genotyping in adenocarcinoma of the uterine cervix in Thailand.

OBJECTIVE: To determine the distribution of human papillomavirus (HPV) genotypes in cervical adenocarcinoma in Thailand and to evaluate the clinicopathologic characteristics associated with common HPV genotypes. METHODS: Formalin-fixed, paraffin-embedded tissues from 150 patients with adenocarcinoma were collected from 4 areas of Thailand. Infection with HPV was detected by nested polymerase chain reaction (PCR) with primers MY09/11 and GP5+/6+. Genotyping was performed using a linear array assay, followed by type-specific PCR targeting the E6/E7 regions of HPV-16, HPV-18, and HPV-52 if the linear array test was negative. RESULTS: Human papillomavirus DNA was detected in 145 (97%) adenocarcinomas (132 single infections; 11 multiple infections; 2 tumors with undetermined HPV type). Genotype 18 was most common (66%), followed by HPV-16 (30%) and HPV-45 (3%). Infection with only HPV-16 and/or HPV-18 accounted for 88% of the HPV-positive tumors. Patients with HPV-18 infection had a younger age (P=0.009) and higher tumor grade (P<0.001) than patients with HPV-16 infection. CONCLUSION: The HPV detection rate in cervical adenocarcinomas in Thailand is high. The predominant genotype is HPV-18, being twice as common as HPV-16. Genotype variations are associated with patient age and tumor grade. Vaccination against HPV-16/HPV-18 might prevent almost 90% of adenocarcinomas.

Association of human papillomavirus type 16 long control region mutation and cervical cancer.

BACKGROUND: The variation of human papillomavirus (HPV) genes or HPV variants demonstrates different risks of cervical cancer. Mutation in the long control region (LCR) at YY1-motifs is one of the mechanisms for enhancing viral oncogene expression during the course of cancer cell progression. In Thai women, cervical cancers are almost always associated with HPV16 variant sub-lineage Asian (HPV16As); however, the mechanism involved remains elusive. The aim of this study was to understand further the oncogenic potential of HPV16As. METHODS: A total of 82 HPV16-positive specimens from Thai women were selected from formalin-fixed paraffin-embedded cervical tissues, and the full length E6 gene of each specimen was amplified and sequenced. LCRs of the HPV16As-positive cases were amplified and sequenced to analyze their polymorphisms. Transcriptional activities of the HPV16As LCRs were then compared with sub-lineage European (EUR), sub-lineage Asian-American 1 (AA1) and HPV16 prototype by insertion of the LCRs into the pGL3-Basic vector. RESULTS: The HPV16 DNA sequences were classified as HPV16 prototype (18.3%), Asian (As, 61%), Asian American-1 (AA1, 8.5%), European (EUR, 7.3%), Asian African-2 (AFR2, 3.7%) and Java-135C (J135C, 1.2%). The prevalence of HPV16As was 30% in low-grade squamous intraepithelial lesion (LSIL), while that in high-grade squamous intraepithelial lesion (HSIL) and squamous cell cervical carcinoma (SCC) were 63.9% and 66.7%, respectively, which demonstrates a significant association of HPV16As with the disease severity. LCR polymorphisms from 43 HPV16As positive cases were analyzed by PCR-sequencing. Thirty-eight nucleotide variation positions spanned nucleotide positions 7157-82. Ten new mutations found in the HPV16As LCRs were located predominantly at the enhancer and proximal to the 3'-end of the early promoter. The LCRs of the common HPV16As, EUR and AA1 showed 5, 13 and 23-fold higher activity than the HPV16 prototype LCR, while those of the new nucleotide variations of As showed 19 (As-sv1) and 30 (As-sv14) -fold higher activity than the HPV16 prototype. CONCLUSIONS: HPV16As DNA sequence variation, especially at the proximal to early promoter in the LCR, enhances transcriptional activity. This could be one of the possible mechanisms for HPV16As-associated cervical cancer development.

Human papilloma virus prevalence, genotype distribution, and pattern of infection in Thai women.

BACKGROUND: The pattern of infection in cervical lesions with respect to HPV subtype has not been systematically studied in Thai women. The aim here was to determine HPV prevalence, genotype, and infection pattern in cervical lesions and to estimate the potential efficacy of an HPV prophylactic vaccine. DESIGN: Formalin-fixed paraffin-embedded cervical tissue blocks of 410 Thai patients from 8 institutes in 4 regions of Thailand (northern, southern, north-eastern, and central) were studied. The samples included 169 low grade squamous intraepithelial lesions (LSILs), 121 high grade squamous intraepithelial lesions (HSILs), and 120 squamous cell carcinomas (SCCs). HPV-DNA was amplified by PCR using consensus primers GP5+ and GP6+. The HPV genotype was then determined by reverse linear blot assay that included 37 HPV-specific 5'-amino-linked oligonucleotide probes. Patterns of infection were classified as single infection (one HPV type), double infection (two HPV types), and multiple infection (three or more HPV types). RESULTS: The mean age of the subjects was 42 years. The prevalence of HPV infection was 88.8%. The highest HPV prevalence was found in the southern region (97.1%) and the lowest in the central region (78.6%). HPV-DNA was detected in 84.6% of LSILs, 90.1% of HSILs, and 93.3% of SCCs. A total of 20 HPV genotypes were identified. The five most common high risk HPV were HPV16 (83.2%), HPV18 (59.3%), HPV58 (9.3%), HPV52 (4.1%), and HPV45 (3.8%). In double and multiple infection patterns, the most common genotypes were HPV16/18 (27.8%) and HPV11/16/18 (54.9%). HPV6 was found only in LSIL and never in combination with other subtypes. HPV11 was most common in LSIL. CONCLUSION: There is no difference of HPV type distribution in women from 4 regions of Thailand with prominent HPV16 and HPV18 in all cases. The bivalent and quadrivalent vaccines have the potential to prevent 48.6 % and 74.5% of cervical cancers in Thai women. The potential of cancer prevention would rise to 87.6% if other frequent HR-HPV types (HPV58, 52, and 45) were also targeted by an HPV vaccine.

Correlation of p53 and Ki-67 (MIB-1) expressions with clinicopathological features and prognosis of early stage cervical squamous cell carcinomas.

AIM: To evaluate the prognostic significance of the expression of p53 and Ki-67, and their correlation with various clinicopathological factors in patients with squamous cell carcinoma of the uterine cervix treated by radical hysterectomy. METHODS: Two hundred and thirty five patients diagnosed between 1987 and 2004 were investigated for p53 and Ki-67 expression by immunohistochemistry. The relationship of these proteins and other potential prognostic factors with recurrence-free survival (RFS) was evaluated. RESULTS: The mean age of the patients was 43.9 years (range, 27-68). There were 28 cases with stage IA2 (11.9%) and 207 cases with stage IB1 (88.1%). The overall 5-year RFS was 91.1% (95% confidence interval [CI] 86.0, 94.4). p53 and Ki-67 expressions were seen in 33.6% and 81.3% of cases, respectively. p53 expression was significantly associated only with parametrial or marginal involvement (P = 0.005), while Ki-67 expression significantly correlated only with larger tumor (P = 0.011) and parametrial or marginal involvement (P = 0.024). There was significant correlation between the expression of p53 and Ki-67 (P = 0.021). In multivariate analysis, Ki-67 expression (hazard ratio for >or=3rd vs 1st tertile [>or=30% vs <20% expression] = 16.0, 95%CI 2.1-124.5; P = 0.0015) appeared to be an independent prognostic factor for RFS, while no prognostic significance of p53 expression was found. CONCLUSION: Ki-67 expression is an independent prognostic parameter for RFS in patients with early stage squamous cell carcinoma of the uterine cervix. Ki-67 expression at or above the cut-off point of 30.0% had significantly poorer RFS.

A variety of atypical manifestations in giant cell arteritis.

Giant cell arteritis (GCA) is a chronic granulomatous vasculitis of large and medium size vessels in the elderly. A new-onset headache is the most frequent symptom. An anterior ischemic optic neuropathy (AION) is one of the most common causes of permanent visual loss. There are four cases with unusual presentation of giant cell arteritis, scalp abscess, prolonged transient monocular visual loss (TMVL), bilateral central retinal artery occlusion (CRAO), and chronic ear pain. All patients had pathologically proven giant cell arteritis, and three of them progressed to blindness in the end. Scalp abscess is a rare sign in GCA. Delay in diagnosis because confusion of the abscess after scalp ischemia with other cutaneous lesions may result in death. TMVL is the forewarning symptom of AION or CRAO in GCA. Early recognition of TMVL is important to make early diagnosis of GCA to prevent blindness. Spontaneous ear pain is extremely rare, and reports have documented delay in diagnosis of GCA resulting in irreversible blindness.

Predictors of the residual disease of high-grade lesions and microinvasive squamous cell carcinoma of the cervix following conization.

OBJECTIVE: To determine the predictors of residual disease of high-grade lesion (HGL) and microinvasive squamous cell carcinoma of the cervix (MICA) in subsequent hysterectomy following conization. MATERIAL AND METHOD: The medical records of women who underwent any conizations diagnosed of HGL and MICA and followed by subsequent hysterectomy within 6 months were retrospectively reviewed. A case and control was defined as whether or not a residual disease of HGL or more was detected in cervical tissue from hysterectomy after conization. Demographic characteristics and pathological features of cases and controls were recorded independently and blindly. Univariate and multivariate analysis were used. The Receiver Operating Characteristics curve of predictors was created using the fitting value obtained from a logistic regression model. RESULTS: A total of 185 women were diagnosed during January 1, 1997 and July 31, 2008 including 102 women without a residual disease and 83 with residual disease at cervical tissue from hysterectomy. The multivariate analysis showed that postmenopausal status (OR = 3.5, 95% CI = 1.8-6.7), number of quadrant involvement (OR = 3.8, 95% CI = 1.8-8.3), internal margin involvement (OR = 3.8, 95% CI = 1.7-8.2), severe nuclear atypia (OR = 2.0, 95% CI = 1.1-3.8) and high mitotic activity (OR = 2.1, 95% CI = 1.1-3.7) were the predictors of residual disease in hysterectomy specimens after conization. Three or more predictors involved predicted the detection of residual disease. CONCLUSION: The presence of postmenopause, three or four quadrants involved, positive internal margin, severe nuclear atypia and high mitotic activity could be used to predict residual lesions after conization.

Primary non-Hodgkin,s lymphoma of the uterine cervix.

Primary malignant lymphoma of the cervix is a rare disease. Because the number of reports of this cancer is limited, there is no consensus on its management, prognosis or the efficacy of various treatments. Primary malignant lymphoma of the cervix stage Ib was diagnosed in a 25-year-old woman. The patient was treated with 6 courses of CHOP chemotherapy (cyclophosphamide, doxorubicin, vincristine, and prednisone). Clinical and pathological responses were complete. This case supports current thinking in that, in selected young patients with primary malignant lymphoma of the cervix who desire to preserve fertility and ovarian functions, combination chemotherapy regimens such as CHOP are the treatment of choice.

HPV genotyping in cervical cancer in Northern Thailand: adapting the linear array HPV assay for use on paraffin-embedded tissue.

OBJECTIVES: The aims of this study were to determine the prevalence of HPV infection and distribution of HPV genotypes in Northern Thai women and thereby estimate the benefit of administering the HPV vaccine in the population. METHODS: Formaldehyde-fixed, paraffin-embedded samples of invasive squamous cell carcinoma from 99 patients were tested for HPV genotypes using the Linear Array HPV Genotyping Test. RESULTS: HPV was detected in 96/99 (96.9%) cases. Seventy-five (78.1%) cases were single infections and 21 (21.9%) multiple. HPV16 and HPV18 were the most common subtypes, detected in 62/96 (64.4%) cases. HPV52 and HPV58 infections were found in 17/96 (17.7%) cases. Co-infection always involved HPV16. The most common co-infection was HPV16 and 52 (7 cases) but never HPV16 and 18. CONCLUSIONS: Although the prevalence of HPV infection in cervical cancer of Northern Thai women is comparable to the other regions worldwide, the distribution of HPV subtypes differs with lower frequencies of HPV16 and 18, and higher frequencies of HPV52 and 58. Moreover, multiple infections are common. The vaccine against HPV16 and HPV18 can be estimated to prevent approximately two thirds of the cervical cancer cases in Northern Thailand. Although designed for use on unfixed tissue, this study shows that the Linear Array HPV Genotyping Test can be successfully used for HPV genotyping on paraffin-embedded archival tissue. This methodology also provides a means for retrospective studies on serial samples for a greater understanding of HPV genotypes, co-infections, and relationship to cervical cancer.

Interobserver reproducibility in determining p16 overexpression in cervical lesions : use of a combined scoring method.

OBJECTIVES: To evaluate interobserver reproducibility of a combined scoring method for immunohistochemical interpretation of p16 overexpression in cervical lesions. MATERIALS AND METHODS: p16 immunostaining was performed in cervical samples from 183 patients, including 69 normal, 42 low grade squamous intraepithelial lesions(LSIL), 36 high grade SIL (HSIL), and 36 squamous cell carcinomas(SCCAs). Each case was evaluated by a combined scoring method based on the percentage of positive cells (score 0-3), the intensity of staining (score 0-3), and the distribution pattern (score 0-2). Immunoexpression for p16 was considered as positive when the combined score was 4-8 and negative with a score of 0-3. Ten pathologists with varied experience in interpretating p16 immunostains evaluated each slide independently. RESULTS: All normal cervical squamous epithelia (69/69) were uniformly negative for p16. All HSILs (36/36), all SCCAs (100/100), and all but one of the LSILs (40/41, 97.6%) showed positive expression. In 172 of 183 cases (93.9%), p16 interpretation was concordant with all pathologists. Eleven cases with discordant results included 10 LSILs and 1 normal mucosa sample. Percentage of agreement of each pathologist pair ranged from 96.7-100% (mean 98.1%) with mean kappa value of 0.96 (range 0.93-1.000). CONCLUSION: The proposed combined scoring method shows good reproducibility among the participating pathologists and good correlation with the histologic diagnosis. This method may be a useful guide in the interpretation of p16 expression in cervical epithelial lesions.

Association between prognostic factors and disease-free survival of cervical cancer stage IB1 patients undergoing radical hysterectomy.

This study was designed to identify prognostic factors of patients with cervical cancer stage IB1 undergoing radical hysterectomy. The medical records and specimens of two hundred and five patients with cervical cancer stage IB1 undergoing radical hysterectomy at Songklanagarind Hospital from July 1995 to June 2005 were reviewed. Patients' age, tumor size, histologic type, tumor grade, depth of invasion, degree of stromal invasion, lymph-vascular space invasion (LVSI), surgical margin status, pelvic node status, and adjuvant treatment were assessed for correlation with disease-free survival (DFS). The mean age of these patients was 44.2 years and the median follow up was 56 months. Twenty five patients (12.2%) developed recurrent disease. The overall 5-year DFS was 86%. In univariate analysis, depth of invasion, degree of stromal invasion, LVSI, and pelvic node status were significant prognostic factors. In multivariate analysis, degree of stromal invasion remained the only independent prognostic factor. In conclusion, degree of stromal invasion was the main independent predictor of prognosis in surgical cases of cervical cancer stage IB1.

Detection of human papillomavirus in intraepithelial lesions and carcinoma of the cervix uteri in southern Thai women.

OBJECTIVE: To evaluate the prevalence of high-risk type human papillomavirus (HR-HPV) in preneoplastic lesions and invasive squamous cell carcinoma (SCC) of the cervix uteri in southern Thai women. MATERIALS AND METHODS: A total of 148 formalin-fixed, paraffin-embedded blocks of cervix tissue were retrieved from the files of the Department of Pathology, Prince of Songkla University Hospital. They were classified as negative for intraepithelial lesion (NIL) in 37 cases, low grade lesion (LGL) in 58 cases, high grade lesion (HGL) in 39 cases and SCC in 14 cases. HR-HPV DNA was tested with an Amplicor HPV (Roche Diagnostics) detection kit. RESULTS: Of the 111 cases, 42 of 58 LGLs (72.4%), 34 of 39 HGLs (87.2%) and 13 of 14 SCCs (92.9%) were positive for HR-HPV DNA. In 37 cases of histologically normal cervix, there were 15 cases that showed the presence of HR-HPV DNA. Applying the HR-HPV results for NILs to the general population, the age standardized incidence rate of HR-HPV infection in the normal Thai population was 12.8%. CONCLUSION: HR-HPV DNA can be found in all grades of intraepithelial lesions and carcinoma of the cervix uteri, even in the histologically "normal" looking cervix. These results provide strong evidence for a role in carcinogenesis of the cervix uteri and the existence of a non-productive or latent period of HPV infection.

Preventive and curative effects of curcumin on the development of gastric inflammatory diseases in rats.

Preventive and curative effects of curcumin on experimental acute and chronic gastric ulcers were investigated to validate its clinical application on a remedy for peptic ulcer. Intraduodenal administration of curcumin, 5-20 mg/kg, inhibited gastric acid secretion in pylorus-ligated rats, and oral administration prevented ethanol-induced acute gastric mucosal lesions. Curcumin (20-80 mg/kg, p.o.) dose-dependently prevented both serotonin-induced gastric mucosal lesions and compound 48/80-induced gastric mucosal lesions in rats. Furthermore, oral administration of curcumin, 10-80 mg/kg, twice daily for 10 days, significantly accelerated the healing of acetic acid-induced chronic gastric ulcer and promoted mucosal regeneration in the ulcerated portion in a dose-related manner. Cimetidine prevented the formation of ethanol-induced gastric mucosal lesions, but not of serotonin-induced and compound 48/80-induced gastric mucosal lesions. Consecutive administration of cimetidine showed a marked acceleration in the healing of acetic acid-induced ulcer. Aminoguanidine, an inducible nitric oxide synthase (iNOS) inhibitor, showed anti-ulcerogenic effects similar to those oberved for curcumin. The present results indicate that curcumin exhibits gastric cytoprotection in the acute lesion models and ulcer healing promotion in the chronic ulcer model. The preventive and curative effects of curcumin might be due to an increase in the mucosal defensive mechanism through its antioxidant property and inhibition of NO or cytokine-mediated inflammation.

Congenital and infantile nephrotic syndrome in Thai infants.

Congenital and infantile nephrotic syndrome reported from the Eastern world is rare and might be a different entity from that in the West. In a retrospective review of 10 nephrotic syndrome in Thai infants (5 girls and 5 boys), 7 were diagnosed with congenital nephrotic syndrome and 3 with infantile nephrotic syndrome. Two had congenital nephrotic syndrome secondary to congenital syphilis. All had edema, ascites, and failure to thrive. Of the 3 patients tested for thyroid function, all showed hypothyroidism. Two patients developed renal failure. Renal tissue was examined from 4 patients from 3 biopsies and 2 autopsies; only 1 patient showed tubular microcysts. Symptomatic therapy was performed concurrently with penicillin therapy in 2 patients having congenital syphilis. Prednisolone, cyclophosphamide, captopril, and enalapril were tried in some patients, with little effect. Five patients died from respiratory failure complicated by later infection, 1 patient died from renal failure, and 4 patients were lost to follow-up. Nephrotic syndrome in the first year of life in the Eastern world is rare. Prognosis of nephrotic syndrome in Thai infants at this time is still poor.

Isolated corneal intraepithelial neoplasia.

OBJECTIVES: To report two cases of isolated corneal intraepithelial neoplasia. MATERIAL AND METHOD: The present study included 2 cases presenting with isolated corneal intraepithelial neoplasia. Both patients were treated by corneal epithelial scraping. RESULTS: The authors present the clinical picture and histopathological findings of 2 patients suffering from corneal intraepithelial neoplasia that did not arise from the corneoscleral limbus. Both patients showed no recurrence after treatment for a period of time. CONCLUSION: Isolated corneal intraepithelial neoplasia is rare. Corneal scraping may be an effective treatment.