Cytological nuclear atypia classification can predict prognosis in patients with endometrial cancer.

OBJECTIVE: Endometrial cancer is one of the leading causes of malignancy in females. Nuclear findings are important for patients with cancer, and can provide valuable information to treating oncologists. We investigated whether nuclear findings were a useful prognostic factor in patients with endometrial cancer. METHOD: We investigated 71 cases of endometrial carcinoma with paired histology and cytology at Kurume University Hospital. We classified endometrial endometrioid adenocarcinoma (EEC) G1 and G2 as type I carcinomas, and uterine papillary serous carcinoma (UPSC), clear cell carcinoma (CC) and EEC G3 as type II carcinomas. For the establishment of the cytological nuclear atypia classification, we examined the following nuclear factors on the cytological smears: mitotic figures, prominent nucleoli, nuclear area and anisonucleosis. RESULTS: There was a significant difference in mitotic figures (P < 0.001) and anisonucleosis (P = 0.026) in cytological smears between type I and type II carcinomas. Based on these findings, we categorized cytological nuclear atypia into three groups, nuclear atypia-1 (57.7%), nuclear atypia-2 (19.7%) and nuclear atypia-3 (22.5%), and this classification system correlated well with prognosis in patients with endometrial cancer (P < 0.001). Furthermore, this classification system was able to extract patients with a good prognosis from those with high-grade carcinomas, such as UPSC+CC+EEC G3, and patients with a poor prognosis from those with EEC G1. CONCLUSIONS: Our system of cytological nuclear atypia classification based on endometrial cytology can predict patient prognosis. Cytological nuclear atypia classification and histological typing may be useful for the treatment and follow-up of patients with endometrial cancer, and should be routinely incorporated into cytological reports.

Association of PD-L1 overexpression with activating EGFR mutations in surgically resected nonsmall-cell lung cancer.

BACKGROUND: Recent clinical trials have shown that immune-checkpoint blockade yields a clinical response in a subset of individuals with advanced nonsmall-cell lung cancer (NSCLC). We examined whether the expression of programmed death-ligand 1 (PD-L1) is related to clinicopathologic or prognostic factors in patients with surgically resected NSCLC. PATIENTS AND METHODS: The expression of PD-L1 was evaluated by immunohistochemical analysis in 164 specimens of surgically resected NSCLC. Cell surface expression of PD-L1 in NSCLC cell lines was quantified by flow cytometry. RESULTS: Expression of PD-L1 in tumor specimens was significantly higher for women than for men, for never smokers than for smokers, and for patients with adenocarcinoma than for those with squamous cell carcinoma. Multivariate analysis revealed that the presence of epidermal growth factor receptor gene (EGFR) mutations and adenocarcinoma histology were significantly associated with increased PD-L1 expression in a manner independent of other factors. Cell surface expression of PD-L1 was also significantly higher in NSCLC cell lines positive for activating EGFR mutations than in those with wild-type EGFR. The EGFR inhibitor erlotinib downregulated PD-L1 expression in the former cell lines but not in the latter, suggesting that PD-L1 expression is increased by EGFR signaling conferred by activating EGFR mutations. A high level of PD-L1 expression in resected tumor tissue was associated with a significantly shorter overall survival for NSCLC patients. CONCLUSIONS: High expression of PD-L1 was associated with the presence of EGFR mutations in surgically resected NSCLC and was an independent negative prognostic factor for this disease.

Effect of hyaluronan tetrasaccharides on epidermal differentiation in normal human epidermal keratinocytes.

OBJECTIVE: Hyaluronan (HA) plays a role in keratinocyte proliferation and differentiation. In addition, HA has been shown to have different biological activities depending on its molecular weight. It has been reported that HA-mediated CD44 activation regulates keratinocyte differentiation. Therefore, the aim of this study was to investigate the influence of HA tetrasaccharides (HA4) on the regulation of keratinocyte differentiation, CD44 gene expression and CD44-phosphorylated protein in human keratinocytes, and compare HA4 with high molecular weight HA. METHODS: Normal human epidermal keratinocytes (NHEKs) were treated at doses of 1 µg mL(-1) HA or HA oligosaccharides (HA4). After treatment, cell viability was checked using an MTT (3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Each differentiation marker and CD44 mRNA expression was detected by real-time PCR. Each differentiation marker and CD44-phosphorylated protein was assessed by Western blotting. RESULTS: Hyaluronan and HA4 showed no cytotoxicity up to a dose of 1 µg mL(-1) . On day 3 after HA4 treatment, each differentiation marker mRNA and K10 protein level was higher than that of the control. On day 9, late differentiation marker mRNA and protein levels were increased with HA and HA4 treatment. In addition, HA4 treatment increased the expression of CD44 mRNA, CD44-phosphorylated protein and intracellular calcium concentrations. HA4 enhanced keratinocyte differentiation and increased CD44-phosphorylated protein levels. CONCLUSION: HA4 may induce epidermal differentiation through phosphorylation of CD44.

Expression of activated signal transducer and activator of transcription-3 predicts poor prognosis in cervical squamous-cell carcinoma.

BACKGROUND: Stat3 is a member of the Janus-activated kinase/STAT signalling pathway. It normally resides in the cytoplasm and can be activated through phosphorylation. Activated Stat3 (p-Stat3) translocates to the nucleus to activate the transcription of several molecules involved in cell survival and proliferation. The constitutive activation of Stat3 has been shown in various types of malignancies, and its expression has been reported to indicate a poor prognosis. However, the correlation between the constitutive activation of Stat3 and the prognosis of cervical cancer patients has not been reported. METHODS: The immunohistochemical analysis of p-Stat3 expression was performed on tissues from 125 cervical squamous-cell carcinoma patients who underwent extended hysterectomy and pelvic lymphadenectomy, and the association of p-Stat3 expression with several clinicopathological factors and survival was investigated. RESULTS: Positive p-Stat3 expression was observed in 71 of 125 (56.8%) cases and was significantly correlated with lymph node metastasis, lymph vascular space invasion, and large tumour diameter (>4 cm) by Fisher's exact test. Kaplan-Meier survival analysis showed that p-Stat3 expression was statistically indicative of a poor prognosis for overall survival (P=0.006) and disease-free survival (P=0.010) by log-rank test. CONCLUSION: These data showed that p-Stat3 expression in cervical cancer acts as a predictor of poor prognosis.

Clinical and histologic features of chronic hepatitis C virus infection after blood transfusion in Japanese children.

OBJECTIVE: To characterize the clinical and histologic features of chronic hepatitis C virus (HCV) infection after blood transfusion in Japanese children. STUDY DESIGN: We studied 231 children with a history of blood product transfusion. Patients were divided into two groups: 116 patients with a history of malignant disease (group 1), 115 patients who had undergone open heart surgery (group 2). We examined changes in serum alanine aminotransferase (ALT) activity and HCV markers, and patients' clinical course. Moreover, in 38 patients in whom the time of HCV infection could be defined, we examined liver histology. RESULTS: The proportions of patients in each group who were anti-HCV-positive were 35 out of 116 (30%) and 20 out of 115 (17%), respectively. Of the anti-HCV-positive patients, the proportions of HCV RNA-positive patients in each group were 30 out of 35 (86%) and 12 out of 20 (60%), respectively. Levels of ALT activity in patients with HCV infection varied widely for several years after blood transfusion; thereafter ALT activity fell to <100 IU/L in 2 groups. Serum ALT activity in patients who were HCV RNA-negative became normal. With regard to liver histology, there were no differences in the grade of necroinflammation or stage of fibrosis in patients with different durations of infection or when patients were analyzed according to the presence or absence of malignant disease. Patients mostly had grade 2-4 inflammation and stage 1-2 fibrosis. Thus, chronic hepatitis C was a morphologically mild disease in most children in this study. CONCLUSIONS: Sixty percent to 80% of children with HCV infection in this study developed chronic hepatitis C. However, examination of liver histology findings in children with chronic hepatitis C showed only mild changes.

Experimental regulation of granulomatous reactions around Schistosoma japonicum eggs implanted into the liver of mice.

Using a liver model, various granulomatous responses against Schistosoma japonicum eggs were studied in C57BL/6 mice immunized with tissue-extracted eggs prior to challenge implantation with freshly laid eggs. In mice receiving two ip injections of 20,000 eggs, there was little effect on early granuloma formation. Three weeks after implantation, however, tissue reaction accompanied by marked fibrosis was significantly augmented, compared to that in the untreated mice. In contrast, when mice were given four ip injections, the early reaction was accelerated and the subsequent fibrosis came to an end earlier than in the twice immunized or untreated mice. Different routes of injection produced differing effects on 2-week granulomas, with an augmented reaction following two sc injections and a diminished reaction following the same number of ip injections. Histologically, the diminished reaction was characterized by less cellularity, especially in the case of eosinophil infiltration.

High prevalence of hepatitis C virus infection in schistosomiasis japonica patients associated with hepatocellular carcinoma.

Schistosomiasis japonica (SCJ) patients frequently develop hepatocellular carcinoma (HCC). This study investigated relationship between SCJ infection, hepatitis virus infection, and incidence of HCC, in 25 patients with chronic SCJ infection and HCC (SCJ with HCC group), 51 patients with chronic SCJ infection without HCC (SCJ group) and 65 HCC patients without SCJ (HCC group). Number of patients who were positive to HBsAg or hepatitis B virus DNA were 4 (16.0%) in the SCJ with HCC group, none (0%) in the SCJ group, and 5 (7.9%) in the HCC group; while number of patients who were positive to anti-hepatitis C virus antibody were 21 (87.5%) in the SCJ with HCC group, 3 (5.9%) in the SCJ group, and 58 (84.6%) in the HCC group. Biopsy was performed for all patients, and background histological features of each specimen were evaluated based on the histological activity index scoring system. Mean scores of inflammatory changes in both the portal area and parenchyma in the SCJ with HCC group were significantly higher than those in the SCJ group. Nodular formation which is common in post-viral hepatitis was frequently observed in the SCJ with HCC group, and histological changes in non-cancerous area of the SCJ with HCC group showed the characteristics of chronic viral hepatitis. We conclude that infection of hepatitis virus, particularly hepatitis C virus, affects synergistically on the hepatocarcinogenesis in patients having SCJ infection.

S-100 protein in glands within decidua and cervical glands during early pregnancy.

Strong immunoreactivity with polyclonal S-100 protein antisera and monoclonal S-100 alpha subunit antiserum was found in glandular cells of the decidua basalis and cervical polyps during early pregnancy. Immunoreactive S-100 protein was negative in glandular cells of the endometrium and cervix of nonpregnant women. It was also negative in endometrial hyperplasia and endometrial carcinoma. While the function of S-100 protein is not known, a relationship between humoral factors related to pregnancy and expression of S-100 protein gene is suggested by the results of this study.

Acidic fibroblast growth factor (FGF-1) in the anterior horn cells of ALS and control cases.

The expression and localization of acidic fibroblast growth factor (aFGF; FGF-1) were examined in the spinal cord of patients with amyotrophic lateral sclerosis (ALS) and controls by reverse transcription-polymerase chain reaction (RT-PCR) method and immunohistochemistry. The RT-PCR experiments demonstrated that aFGF amplification products were clearly detected in all control cases but could be scarcely seen in ALS patients. aFGF immunoreactivity was detected in the anterior horn cells of the spinal cord. Double immunostaining for aFGF and choline acetyltransferase revealed that the majority (95.9%) of cholinergic neurons expressed aFGF. In ALS cases, the number and the staining intensity of aFGF-positive neurons were markedly decreased. These results suggest that aFGF is involved in ALS pathology.

Primary structure of GAP-43 mRNA expressed in the spinal cord of ALS patients.

Recent evidence suggests that the growth associated protein GAP-43 is involved in the pathology of amyotrophic lateral sclerosis (ALS). In order to assess the primary structure of the GAP-43 mRNA expressed in the spinal cord of ALS patients, the total coding region of the GAP-43 mRNA was amplified from postmortem human spinal cord specimens using the reverse transcriptase-polymerase chain reaction (RT-PCR) method. GAP-43 amplification products were clearly detected in all of the ALS cases but not in the normal controls. The GAP-43 mRNA and the deduced amino acid sequences from all ALS cases coincided completely with the sequence of human fetal GAP-43. These results suggest that the abnormal expression of GAP-43 mRNA underlying the pathogenesis of ALS is due to a quantitative increase, and that there are no qualitative abnormalities associated with a change in the amino acid sequences.

Gastric enterochromaffin-like-cell tumor with liver and splenic metastases.

We report a patient with gastric enterochromaffin-like-cell tumor with liver and splenic metastases. He was 68 years old and presented with major complaints of epigastric pain and weight loss. Under the diagnosis of gastric carcinoma with liver metastasis, total gastrectomy with splenectomy and lateral segmentectomy of the liver was performed. Intraoperative findings resulted in a diagnosis of adenocarcinoma T3N2P0H1, in stage IVa. Histological examination of the resected specimens showed a well differentiated neuroendocrine carcinoma (enterochromaffin-like-cell tumor) with liver and splenic metastasis which demonstrated high-grade lymphatic and vascular invasion. There was no lymph node metastasis. The tumor cells in the stomach, liver and spleen were immunoreactive for chromogranin A and Grimelius--positive. We review the literature, as well as presenting this case report.

The possible intraspousal transmission of HCV in terms of lichen planus.

Lichen planus (LP), common mucocutaneous disorder, involves not only oral mucosa and skin but genitalia membrane. LP is frequently seen in patients with HCV infection. This study evaluated patients with HCV-associated oral lichen planus (OLP) for vulvar and vaginal LP involvement, and the possible intraspousal transmission of HCV. We examined a total of 24 female Japanese patients with OLP for genitalia LP: 14 OLP-HCV positive and 10 OLP-HCV negative. All subjects were evaluated for genital LP by a gynecologist. All 24 subjects and 10 of the husbands were tested for anti-HCV and serum HCV RNA. Vulvar LP was observed in 10 (41.7%) of 24 patients with OLP. Vulvar LP in 14 (OLP-HCV positive) and 10 patients (OLP-HCV negative) were observed in 42.9 and 40%, respectively. There were no significant differences (age, sites of OLP, blood transfusion, HCV infection, and degree of liver diseases) between the vulvar LP and non-vulvar LP patients. Two spouses of 10 married couples were shown to be infected with HCV. In one couple with HCV infection, the wife and husband had also erosive OLP, the wife had erosive vulvar LP. In conclusion, the majority of OLP patients suffered from genitalia LP in Japan. Clinicians should follow the OLP patients with sufficient attention to the presence of extraoral manifestations. These data may suggest the occurrence of intraspousal transmission of HCV through erosive vulvar LP.

Infantile cholestasis due to cytomegalovirus infection of the liver. A possible cause of paucity of interlobular bile ducts.

We report a neonatal case of hepatic cytomegalovirus (CMV) infection associated with paucity of interlobular bile ducts. The premature infant developed severe jaundice, and died of disseminated CMV infection. The diagnosis of CMV infection was based on detection of pathognomonic inclusions in the liver and on a high titer of CMV antibody in the serum. The hepatic histologic findings in this infant included severe cholestasis, complete absence of interlobular bile ducts in the majority of the portal tracts, and CMV inclusions in the duct epithelium. Interestingly, the CMV inclusions were localized in the bile duct, but not in the parenchyma. We postulate that the paucity of interlobular bile ducts may have been a sequela of CMV infection of the liver arising from destruction of the bile ducts, leading to severe cholestasis.

Different courses of granulomatous reactions around Schistosoma japonicum eggs in three strains of mice.

Tissue reaction around Schistosoma japonicum eggs was studied after implantation into 3 strains of mice. When freshly laid eggs were implanted into the livers of C57BL/6, CBA/J, and BALB/c mice, the tissue reaction, which could be divided into 3 stages, i.e., abscess formation, inflammatory stage, and fibrous stage, differed in persistence and reactivity according to the strain. This was true especially during the later 2 stages. In BALB/c mice, the inflammatory stage ended earlier. In CBA/J mice, it continued for the longest period and the subsequent fibrosis was the most marked. In C57BL/6 mice, the period of persistence of this inflammatory stage was intermediate, but these mice showed the highest degree of inflammatory reaction. When footpad reactions were tested, the delayed-type hypersensitivity reaction was generally comparable with the morphometric analysis. In presensitized mice implanted with lyophilized eggs, tissue reactivity was analogous to the fibrous stage of freshly laid egg-implanted mice. In addition, there was an inverse relationship between antibody level and granuloma size, particularly at later reactions. The analysis of experimental granuloma formation presented in this report contributes to the study of stage-specific regulation.

Inhibitory effect of circulating egg antigens on Schistosoma japonicum egg-induced granuloma formation.

Schistosoma japonicum produces an enormous quantity of eggs during infection. This study was conducted to examine the effect of egg-derived antigens on the development of granuloma formation around S. japonicum eggs in the livers of mice. When soluble egg antigen (SEA) (75 micrograms/mouse/day) was injected 3-4 times via a vein into mice implanted with laid eggs, the magnitude of tissue lesion was drastically inhibited when assessed at maximal occurrence (14 days after implantation of eggs), whereas adult worm antigen (AWA), rabbit hyperimmune serum against SEA, or bovine serum albumin (BSA) did not show any effect on either the cellularity or the magnitude. In contrast to intravenous injection, there was no effect from subcutaneous injections of SEA. When serum taken from heavily infected mice or rabbit was transferred, there was a considerable extent of inhibition. In addition, an immune complex fraction of infected rabbit serum was found to have a stronger inhibitory effect than the supernatant fraction. This study indicates that the amount of egg-derived circulating antigens has a crucial effect on the development of schistosome granuloma formation.

Fatal acute hepatitis B virus infection while receiving immunosuppressants after renal transplantation.

A 47-year-old man with acute hepatitis B virus (HBV) infection who had been receiving immunosuppressants after renal transplantation developed progressive liver failure. During the clinical course (approximately 7 months), anti-HBc IgM antibody and HBV-DNA polymerase levels remained high, but the serum alanine aminotransferase (ALT) level was consistently less than 150 K.U. Histopathologic examination of the liver showed submassive hepatic necrosis without significant inflammation accompanied by marked fibrosis. Most hepatocytes showed strong nuclear expression of HBc antigen by immunohistochemical staining and electron microscopy revealed numerous intranuclear core-like particles. Hepatitis B virus infection in immunosuppressed individuals occasionally insidiously progresses, resulting in liver failure. The clinical course of such patients thus merits close scrutiny.

Budd-Chiari syndrome complicated by hepatocellular carcinoma with no evidence of infection with hepatitis virus: a case report.

Hepatocellular carcinoma occurs in patients with Budd-Chiari syndrome. However, the etiology of hepatocellular carcinoma accompanied with Budd-Chiari syndrome has not been elucidated. We report a case of Budd-Chiari syndrome with membranous obstruction of the inferior vena cava complicated by hepatocellular carcinoma in an 80 year-old man. There was no evidence of co-infection with hepatitis A, B, C, D, E, and G virus. Histologically, the non-cancerous liver tissue showed chronic venous congestion with no evidence of hepatitis virus-associated liver cirrhosis. This case suggests that chronic venous congestion of the liver may be one of the pathologic conditions that occurs in hepatocellular carcinoma.

Ultrastructural studies of the bile duct in alcoholic liver disease.

The fine structural characteristics of the bile duct in patients with alcoholic disease are described. Dark cell metamorphosis, edematous microvilli, and increased number of pinocytotic vesicles on the basal wall surface of the duct epithelium were observed. These alterations may be interpreted as evidence of disordered water metabolism, probably reflecting secretion and reabsorption hyperfunction in the duct epithelium. In addition, widened intercellular spaces in the basal half of the epithelium suggested retention of fluid following reverse pinocytosis along the lateral cell surface. Although no alterations of the duct epithelium distinct from those in patients with other liver diseases were apparent in patients with alcoholic liver disease, the basement membrane of the bile duct exhibited unusual duplication with multiple layers and occasional loop-formation in lacunae on the basal surface.