Identification of serum miRNAs as novel non-invasive biomarkers for detection of high risk for early gastric cancer.

BACKGROUND: Many micro-RNAs (miRNAs) are differentially expressed in Helicobacter pylori-infected gastric mucosa and in gastric cancer tissue and previous reports have suggested the possibility of serum miRNAs as complementary tumour markers. The aim of the study was to investigate serum miRNAs and pepsinogen levels in individuals at high risk for gastric cancer both before and after H. pylori eradication. METHODS: Patients with recent history of endoscopic resection for early gastric cancer and the sex- and age-matched controls were enrolled. Serum was collected from subjects before or after eradication and total RNA was extracted to analyse serum levels of 24 miRNAs. Serum pepsinogen (PG) I and II levels were measured using enzyme-linked immunosorbent assay kits. RESULTS: Using miR-16 as an endogenous control, the relative levels of miR-106 and let-7d before and after H. pylori eradication and miR-21 after eradication were significantly higher in the high-risk group than in the controls. H. pylori eradication significantly decreased miR-106b levels and increased let-7d only in the control group. After eradication, the combination MiR-106b with miR-21 was superior to serum pepsinogen and the most valuable biomarker for the differentiating high-risk group from controls. CONCLUSION: Serum miR-106b and miR-21 may provide a novel and stable marker of increased risk for early gastric cancer after H. pylori eradication.

Toxoplasma gondii does not persist in goldfish (Carassius auratus).

Recent reports of toxoplasmosis in marine mammals raise concern that cold-blooded marine animals are a potential source of Toxoplasma gondii infection. To examine the transmissibility of T. gondii to fish, we observed the development of T. gondii tachyzoites inoculated into oviduct epithelial cells of goldfish (Carassius auratus) microscopically in vitro. Further, the survival period of tachyzoites inoculated into goldfish muscle was bioassayed in mice and through PCR analysis. In cell cultures at 37 C, both RH and Beverley strains of T. gondii tachyzoites had penetrated into cells at 6 hr post inoculation, and were multiplying. In cell cultures at 33 C, many tachyzoites of both strains attached to the host cells, but no intracellular tachyzoites were observed at 24 hr post inoculation. In the T. gondii inoculated goldfish kept at 33 C, tachyzoite DNA was detected in the inoculated region on day 3, but not on day 7. When inoculated goldfish were kept at 37 C, live tachyzoites were seen at the inoculation site on day 3, but not on day 7. These results suggest that T. gondii does not persist in fish.

Isolated posterior cruciate ligament insufficiency induces morphological changes of anterior cruciate ligament collagen fibrils.

We studied the ultrastructural changes of the human anterior cruciate ligament (ACL) with transmission electron micrograph cross-sections following isolated posterior cruciate ligament (PCL) injury. Biopsy specimens were obtained from the proximal third and anteromedial aspect of the ACL. Fourteen patients with PCL-deficient knees at a mean of 22.1 months from injury to surgery and 5 normal knees amputated secondary to malignant tumors or traumatic injuries were used as controls. A significant difference was found in the number of collagen fibrils per 1 microm2 between the PCL-deficient knee group and the control group. There was a significant difference found in the collagen fibril diameter between the PCL-deficient knee group and the control group. The collagen packing density (the percentage of sampled area occupied by collagen fibrils) was also significantly different between the PCL-deficient knee and the control group. The current study shows that an isolated PCL insufficiency can induce morphological changes in ACL collagen fibrils, suggesting that a PCL insufficiency can have adverse effects on other ligamentous structures in the knee joint.

Image storing system for radiation therapy (radiation oncology information system: ROIS) as a branch of diagnostic PACS; implementation and evaluation.

We have implemented the new system that is called as ROIS for storing medical images of the patients underwent radiotherapy as a branch of medium-scale diagnostic PACS. It was estimated that, for about nine years, we can store all medical images of the patients who received radiation therapy under the on-line storage condition. Further, we can retrieve all medical images of the radiotherapy patients on ROIS faster than conventional film library or on the diagnostic PACS, even if the patient had been treated many years ago. Implementation of ROIS provided us a great help in our clinical work.

The adverse effects of sectioning the posterior cruciate ligament in rabbits. Changes in the structural and morphological properties of the femur-anterior cruciate ligament-tibia complex.

This study examined the changes in the structural properties of the femur-anterior cruciate ligament-tibia complex (FATC) and the histologic changes of the anterior cruciate ligament (ACL) following sectioning of the posterior cruciate ligament (PCL) of 20 rabbits. The PCL in the right knee was sectioned through an arthrotomy. The left knee underwent arthrotomy only and was used as a control. The animals were killed 3 and 6 months postoperatively. The tensile properties of the FATCs were tested, and the ACLs were histologically examined using polarized light microscopy and transmission electron microscopy. There were significant decreases in the ultimate load following sectioning of the PCL, although there were no significant changes in the stiffness. There were no significant differences in either the crimp period or the crimp amplitude of the ACL following sectioning of the PCL. There were significant increases in the number of collagen fibrils per square micrometer, and significant decreases in the collagen fibril diameter and proportion of total collagen fibril area per square micrometer following sectioning of the PCL. These findings suggest that isolated PCL injury may cause pathological changes in the ACL and its insertion sites.

Peripheral primitive neuroectodermal tumour with ganglioneuroma-like areas arising in the cauda equina.

Peripheral primitive neuroectodermal tumour (pPNET or peripheral neuroepithelioma) is one of the malignant small round cell tumours of peripheral nerves, soft tissues and bones, but rarely originates in the spinal canal. We report an example of pPNET arising in the cauda equina of a 14-year-old Japanese boy. At surgery, a well-demarcated tumour measuring 2 x 4 cm in diameter and involving one of the nerve roots of the cauda equina was located within the intradural space with no evidence of extradural extension. Microscopically the tumour was made up of sheets of closely packed small round cells, associated with ganglioneuroma-like islands. Immunohistochemically, the small round tumour cells were intensely positive for neuron-specific enolase (NSE), an MIC2 gene product (O13) and beta 2-microglobulin, whereas the foci with ganglion cell-like cells reacted positively to NSE, synaptophysin and beta 2-microglobulin but were negative for O13. A chimeric transcript of the EWS/FLI-1 fusion gene detected by a nested reverse transcriptase-polymerase chain reaction using formalin-fixed paraffin-embedded tissue justified the diagnosis of pPNET. Only 6 cases of PNET in the cauda equina have been described in the literature, and this is the first case of a pPNET with ganglio-neuroma-like areas. This finding suggests that the primitive tumour cells of pPNET may respond to unknown inductive effects and express a ganglion cell-like morphology.

18 years of conformation radiotherapy at Nagoya University Hospital.

Conformation radiotherapy is one of the best techniques for minimizing the radiation dose absorbed by the surrounding normal tissue while delivering a high dose to a cancerous target area. The cases of all patients who underwent external irradiation at Nagoya University Hospital from 1975 to 1992 were reviewed. A total of 5740 patients with 6179 lesions were irradiated during this time, and 3795 treatment plans involved radical intended irradiation. Of the 5740 patients, 1017 had head and neck cancer, 982 had cervical cancer, 506 had lung cancer, 439 had primary brain tumors, 308 had esophageal cancer, 1213 had metastatic tumors, and 1275 had other types of tumors. The total number of treatment plans per year decreased from 442 in 1975 to 292 in 1992. Likewise, the percentage of conformation radiotherapy performed in all patients decreased from 29.4% (130/442) in 1975 to 8.6% (25/292) in 1992. It occupied 14.5% (982/6179) of all intended plans, and 20% (775/3795) of radical treatment plans. The conformation technique was used in cases of cervical cancer (72%), esophageal cancer (65%) and primary brain tumors (25%). Boost Conformation radiotherapy represented 2% of all treatment planning and 29% of the conformation radiotherapy. Boost Conformation radiotherapy has recently become more popular and now represents more than 50% of conformation radiotherapy. With respect to cases of cervical cancer, the rates of local recurrence and late complications in cases treated by conformation radiotherapy were lower than in cases treated by two parallel opposed radiotherapy.

[Influence of the overall time of radiotherapy on local control for T2 cancer of the tongue].

Forty patients with T2 cancer of the tongue treated with external irradiation plus radium implants at Aichi Cancer Center were reviewed retrospectively to analyze the influence of overall treatment time on local control. One hundred percent (11/11) local control was achieved in patients irradiated within 35 days, in contrast to only 52% (13/25) local control in those irradiated over 35 days. This result indicates that prolonged overall time is associated with loss of local control. This prolongation is mainly due to the prolonged interval between external and interstitial irradiation. The interval was likely to be longer because of enhanced acute radiation mucositis, especially in patients administrated bleomycin during external irradiation. External irradiation should be followed by interstitial irradiation as soon as possible to keep overall time shorter.

[A retroperitoneal embryonal carcinoma with a high HCG level].

A case of an embryonal carcinoma of the retroperitoneum in a 23-year-old male is reported. His serum, urinary HCG, serum alpha-fetoprotein, and LDH values were high. After combination chemotherapy, when the tumor markers decreased to within normal values, the retroperitoneal tumor was extirpated. Histologically, a small focus in the embryonal carcinoma and cartilage was seen in the fibrous scar tissue. Immunohistochemically, the cancer cells revealed negativity for HCG, alpha-fetoprotein, and the epithelial membrane antigen. However, with the recurrence of the tumor, his HCG and LDH values elevated. The patient died 8 months after the surgical extirpation of his retroperitoneal tumor.

[Studies on relationship between histology, tumor markers (prostatic acid phosphatase.prostate specific antigen.gamma-seminoprotein.leu-7) and clinical course in prostate cancer].

We are interested in the therapeutic response to chemotherapy and radiotherapy of relapsed prostate cancer. In 9 cases of prostate cancer treated by endocrine therapy, tumor markers (PAP.PA.gamma-Sm.Leu-7) and cell types at the start of endocrine therapy and that taken at a hormone independent point were compared between prostatic tissue obtained. All cases had a period of response to endocrine therapy, but subsequently relapsed. The results were divided into the following 3 groups: Group I (changed cell type.decreased positive rate of markers) had the shortest response duration to endocrine therapy and there was no response to chemotherapy; Group II (unchanged cell type.decreased positive rate of markers) had a long response duration and slow progression under endocrine therapy; Group III (unchanged cell type.unchanged positive rate of markers) was chemo- or radiotherapy sensitive during post-endocrine therapy relapse. These results suggest that this is an effective method which dictated the choice of treatment method and allowed an approximate prognosis for relapsed prostate cancer previously treated by endocrine therapy.

[A case of adenoid cystic carcinoma of the esophagus].

A surgical case of an adenoid cystic carcinoma (ACC) of the esophagus in a 75-year-old man is reported. Histologically, the tumor consisted of an ACC, a squamous cell carcinoma and a small tubular adenocarcinoma. The ACC and the tubular adenocarcinomatous regions in the submucosa and the lamina propria were continuous with the overlying squamous cell carcinoma and atypical squamous epithelium. Immunohistochemically, two types of tumor cells were detected in the ACC. One was found to have EMA-positive epithelial cells whereas the other had actin-positive myoepithelial cells. In the normal esophageal gland, actin-positive cells are found at the periphery of acini and around the layer of epithelial cells in the small duct but they have not been detected in the main duct. These findings suggest that the tumor developed from the small duct and differentiated into two directions: an ACC and a squamous cell carcinoma.

[A case of small cell carcinoma of the urinary bladder].

A case of small cell carcinoma with a high serum alpha-fetoprotein level of the urinary bladder is reported. Histologically, in addition to the small cell carcinoma, there were small areas of atypical carcinoids and larger epithelial cells. Immunohistochemically, the small cell carcinoma and the atypical carcinoids showed positivity for neuron-specific enolase, chromogranin, and vimentin. On the other hand, the larger epithelial cells showed positivity for alpha-fetoprotein, keratin, and carcinoembryonic antigen. Electron microscopically, a few neurosecretory granules were noted in the cytoplasm of the small cell carcinoma. After a total cystectomy, the serum alpha-fetoprotein level decreased to within a normal range. Postmortem examination of recurrent tumorous masses revealed only features of the small cell carcinoma. This case supports a theory that a small cell carcinoma of the urinary bladder develops from multipotential epithelial cells.

[A case of vesical carcinoma with sarcomatous changes--immunohistochemical and electron microscopic observations].

The immunohistochemical and electron microscopic characteristics of a vesical carcinoma with a sarcomatous change in a 90-year-old man are described. Keratin was strongly positive in the carcinomatous region and was also identified in the sarcomatous region. CEA was present only in the carcinomatous region. Both the carcinomatous and sarcomatous regions were negative for S 100 protein, myoglobin, lysozyme, and desmin. Electron microscopically, a few tumor cells with tonofilaments were observed in the sarcomatous region. Many tumor cells in the sarcomatous region had no characteristic structures suggestive of epithelial or mesenchymal origin. However, transitional types of tumor cells, between the epithelial cells and the cells with neither epithelial nor mesenchymal differentiation, were noticed. Therefore, this suggested that the sarcomatous region was a site of immature epithelial cell proliferation.

[A case of small cell carcinoma of the prostate].

A case of a small cell carcinoma of the prostate that occurred in a 68-year-old man is reported. Needle biopsy of the prostate showed an adenocarcinoma. A second biopsy revealed both an adenocarcinoma and a small cell carcinoma. A subsequent third biopsy revealed only a small cell carcinoma, and the patient died of respiratory failure 26 months after the initial presentation. An autopsy revealed the tumor that had replaced the prostate extended into the bladder and rectum. Widespread metastatic foci, showing a histologic pattern of solely a small cell carcinoma, were present in various organs. The adenocarcinoma component was seen restricted to the prostatic region. Immunoperoxidase staining for prostate-specific antigen, prostate-specific acid phosphatase, gamma-seminoprotein, and leu-7 showed positivity only in the adenocarcinoma, whereas neuron-specific enolase was positive only in the small cell carcinoma.

[An autopsy case of malignant retroperitoneal paraganglioma].

A case of an extra-adrenal malignant paraganglioma of the retroperitoneum is reported. A 33-year-old woman with a complaint of a dull pain in the right thigh was admitted. A CT scan revealed a large retroperitoneal mass situated between the right kidney and the aorta. A biopsied specimen of the right femur uncovered a metastatic focus of a paraganglioma. Subsequent chemotherapy and irradiation ultimately proved ineffective, and the patient died at the age of 37. Autopsy revealed a retroperitoneal mass 11 X 9 X 4 cm, and its metastatic foci in the vertebrae, ribs, sternum, and liver. Six cases of extra-adrenal malignant paragangliomas of the retroperitoneum that have been reported in Japan also are briefly reviewed.

[Multilocular epidermoid cyst of the spleen--report of a case observed with an electron microscope].

A 27-year-old female, complaining of left hypochondralgia, underwent a splenectomy and a partial pancreatectomy. The spleen, 18.5 x 14 x 6 cm in size and weighing 1060 g., was occupied by a large multicystic mass. The cystic cavities contained turbid chocolate-colored fluid or a yellowish creamy substance. The cysts varied in size and were lined by stratified squamous epithelium. By means of an electron microscope, a few Langer hans cells were seen in the stratified squamous epithelium, but melanocytes and Merkel cells were not noticed. Twenty-eight cases of splenic epidermoid cysts reported in Japan are briefly reviewed.

Adenofibroma of the rete testis. A case report with electron microscopy findings.

The light and electron microscopic characteristics of an adenofibroma of the rete testis in a 51-year-old man are described. The tumor was 5.5 cm in greatest diameter and situated in the anterior superior portion of the right testis. It was composed mainly of mesenchymal and secondary epithelial proliferation. Long slit-like spaces and short tubules lined by a layer of epithelial cells were seen in the mesenchymal tissue. The epithelium was histochemically and ultrastructurally similar to that of the rete testis, and the tumor was considered to be of rete testis origin.

[A bronchogenic cyst of the esophageal wall--a case report and differential diagnosis from an esophageal cyst].

A 25-year-old man underwent extirpation of a cystic mass of the esophageal wall. It proved to be an unilocular cyst surrounded by a pseudo-stratified, ciliated columnar epithelium with occasional goblet cells. Outside of the epithelium, a smooth muscle layer was seen and a small foci of seromucous glands were scattered in the wall. Electron microscopic observations of the epithelial layer revealed 5 types of the cells: ciliated columnar cells, mucous cells, brush cells, intermediate cells, and basal cells. Our results suggest that electron microscopic observations are useful for distinguishing the bronchogenic cyst from the esophageal cyst. Nine cases of para-esophageal bronchogenic cysts that have been reported in Japan also are briefly reviewed.