Histochemical techniques have contributed significantly to advances in placental biology and cell biology. In this mini-review, we describe recent advances in histochemical technologies and show how these technologies can profoundly improve our understanding of placenta morphological function related to health and disease. Fundamental theories and applications of five separate methods discussed here are 1) tissue-based polymerase chain reaction by laser microdissection, 2) a novel antigen retrieval method using citraconic anhydride plus heating, 3) immunohistochemical detection of Lewis-related antigen expression and galectin-1 binding in the human placenta, 4) confocal microscopy analysis of IgG transport in placental trophoblasts, and 5) high-resolution immunofluorescence and correlative microscopy using ultrathin cryosections in placental research. This review article is based on a presentation given in a workshop entitled Histochemistry: Theory and Application at the 12th International Federation of Placenta Associations Meeting held in Kobe, Japan, on September 9, 2006.

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The hypothalamic-pituitary-adrenal axis is an essential component for the maintenance of homeostasis following trauma. Major surgical trauma often induces overwhelming inflammatory responses leading to sepsis and organ dysfunction. This study was designed to evaluate the adrenal responses both before and after various degrees of surgical trauma and to determine the incidence of postoperative relative adrenal insufficiency resulting in the marked inflammatory response often associated with postoperative complications. Fifty-one surgical patients were divided into groups who underwent major, moderate, and minor surgeries. Before the operation and during resting conditions, a short corticotropin (ACTH) stimulation test was performed in each patient. The postoperative concentrations of serum cortisol, interleukin (IL)-6, IL-10, C-reactive protein (CRP), and plasma ACTH were measured. Fifty of 51 patients were identified as responders to ACTH. The postoperative cortisol levels were the same as those obtained by ACTH stimulation in highly and moderately stressful surgeries. The increases in postoperative IL-6 and CRP levels were greatest with major surgery, intermediate with moderate surgery, and least with minor surgery. Furthermore, plasma ACTH levels increased after major and moderate surgeries; however, there was no significant differences in postoperative serum IL-10 levels. Systemic inflammatory response syndrome (SIRS) was found in 11 of 17 patients (64.7%) who underwent major surgery and in 4 of 16 patients (25%) who underwent moderate surgery (p=0.037). The duration of SIRS was significantly longer in patients undergoing major surgery (62 ± 20 hrs) than in patients undergoing moderate surgery (21 ± 3 hrs, p=0.038). Postoperative complications were more frequent in patients undergoing major surgery (41.2%) than in patients undergoing moderate surgery (6.3%, p=0.039). Furthermore, there were significant differences in the length of the postoperative stay among the three groups (p<0.01). One nonresponder had serious postoperative inflammatory complications. These results suggest that a short ACTH stimulation test performed preoperatively is a helpful method for determining the maximal cortisol response to surgical trauma and to identify high-risk individuals and that a relative postoperative adrenal insufficiency may be closely related to the decreased cortisol secretion following major surgical trauma.

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In this study, we investigated heat shock protein (HSP) expression and stress fiber (SF) formation in endothelial cells (ECs) within the arterial vascular tree of adult rats under normal physiological conditions. Using quantitative immunofluorescence microscopy, we found no significant differences in expression of HSPs 25, 60, 70, and 90 among ECs in the straight portions of rat arteries. In these regions, ECs appeared spindle-shaped and contained short bundles of central SFs. In contrast, ECs in the curved portions or the branch sites of the arteries, exhibited striking differences in HSP expression. ECs with higher HSP expression were localized at the lesser curvature in the curved portions or the distal site of the branch ostia. Moreover, the ECs became polygonal and contained irregular central SFs at the lesser curvature. At the branch sites, downstream ECs became spear-shaped and contained long, thick bundles of central SFs. Curved portions or branch sites are the regions of disturbed flow at which early atherosclerotic lesions are often found. Our results demonstrate these positional differences in HSP expression associated with changes in SF formation within the arterial vascular tree under non-pathological conditions. Our study provides basic information for understanding stress responses via HSP expression and SF formation in vascular ECs and the pathogenesis of atherosclerotic disease.

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Vertebral artery injury associated with non-penetrating cervical trauma is rare. We report 11 cases of vertebral artery injury diagnosed with magnetic resonance imaging (MRI) after blunt trauma to the cervical spine and discuss about the importance of MRI in the diagnosis of this injury. Seven cases were caused by motor vehicle accidents, three by diving accidents, and one by static compression of the neck. All of the patients had documented cervical spine fractures and dislocations. In three patients, the diagnosis of complete occlusion of the vertebral artery was made on the basis of MRI and digital subtraction angiography (DSA). In the other patients, mural injuries of the vertebral artery were demonstrated with DSA. These 11 patients presented with acute, nonspecific changes in neurological status. Two had infarctions of the cerebellum and brainstem. None were treated with anticoagulants. All of them survived and were discharged to other hospitals for physical and occupational therapy. Although DSA remains the gold standard for diagnosing vertebral artery injuries, MRI is a newer modality for assessing cervical cord injury, and it may be useful for evaluating the presence of vertebral injury after blunt cervical spine trauma.

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Eighty-four patients underwent laparoscopic cholecystectomy (LC) from January through August 2006. Of these patients, 4 (4.7%) were found to have occult gallbladder carcinoma (GC) either during or after the procedure. Two of the patients were women and 2 were men. The mean age was 75.0 years. One patient had mucosal tumors, 2 had subserosal tumors, and 1 had a serosal lesion. One of the 2 patients with subserosal tumors underwent radical surgery. In a previous study, 0.83% (10 of 1,195) of patients who had undergone LC were found to have occult GC, either during of after the procedure. The prevalence of gallbladder carcinoma has recently been increasing. GC has been reported in 0.3% to 1.5% of patients who have undergone cholecystectomy. Since the introduction of laparoscopic surgery, the number of cholecystectomies being performed has increased, which may explain why occult GC seems to be occurring more frequently. The prognosis for GC is poor, and surgical resection is the only potentially curative treatment. However, GC is difficult to diagnose at an early stage and difficult to recognize even in the advanced stages. Fifteen percent to 30% of patients show no preoperative or intraoperative evidence of malignancy. Occult GC is also increasing. Because flat infiltrating GC and GC with cholecystitis and numerous stones are difficult to diagnose preoperatively, we recommend taking frozen sections from patients who are of advanced age (older than 70 years), have a long history of stones, or have a thickened gallbladder wall.

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Although injection-site granulomas caused by leuprorelin acetate have been reported, there have been no reports of granulomas caused by both leuprorelin acetate and goserelin acetate. An 81-year-old man presented with subcutaneous nodules of the abdominal wall and upper arm, where 11.25 mg of leuprorelin acetate had been injected for the treatment of prostate cancer. Because of these nodules, treatment was changed to goserelin acetate. Nevertheless, he presented with another subcutaneous nodule at the injection site. Histological examination showed that these nodules consisted of numerous giant cells that were CD3-positive T lymphocytes and CD68-positive histiocytes associated with granulomatous changes. The granulomas had likely been caused by delayed-type hypersensitivity to leuprorelin acetate injection. The granuloma that formed after goserelin acetate injection might thus have developed owing to the immunogenicity of the previous leuprorelin acetate injections. The patient underwent surgical castration. The present case suggests that both leuprorelin acetate and goserelin acetate can cause injection-site disorders.

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Hand metastases occur infrequently, and metastatic tumors in the soft tissue of the hand caused by rectal cancer are extremely rare. We report a case here. The patient was a 76-year old man. He underwent Miles operation for rectal cancer located in the lower portion of the rectum. Histopathologically, the resected specimen showed well-differentiated adenocarcinoma. Six years postoperatively, a tumor involving the soft tissue of the palma was found in his left hand. The tumor was resected, and pathological examination showed a well-differentiated adenocarcinoma similar to the primary rectal carcinoma. Immunohistochemical examination demonstrated that this hand tumor had metastasized from rectal cancer. Fifteen cases of colorectal metastatic tumors in the hand have been documented, of which three were soft-tissue metastases. This report describes the fourth case.

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A case of hepatocellular carcinoma (HCC) causing a major arterioportal (A-P) shunt is reported. The patient exhibited massive ascites and tested positive for hepatitis B surface antigen. An abdominal computed tomography (CT) examination showed a low-density lesion in the left lobe of the liver and an A-P shunt, but no tumor stain was visible. Upper gastrointestinal endoscopy revealed severe esophageal varices. Because the tumor marker level was abnormally high, an HCC causing an A-P shunt in a cirrhotic liver background with severe esophageal varices as a result of portal hypertension was diagnosed. We performed endoscopic variceal ligation to treat the severe esophageal varices and interventional radiology treatment for the A-P shunt and HCC, but the patients condition was unchanged. Because the patients liver function gradually improved, surgical treatment was selected. The patient underwent left hepatectomy. Pathological examination revealed a poorly differentiated HCC in a cirrhotic liver background. The postoperative course was uneventful, and the patient was discharged 2 weeks after the operation.The patient subsequently underwent transcatheter arterial embolization therapy for recurrent HCC in the right lobe of the liver, but the esophageal varices disappeared.

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Here, we report on a patient with squamous cell carcinoma (SCC) arising from recurrent anal fistula. The patient was a 57-year-old woman who had 32-year history of having a recurrent perianal abscesses that ruptured spontaneously. Six months before her admission to our hospital, anal pain developed. She had no history of inflammatory bowel disease. Physical examination revealed three external fistulous openings at the two oclock position, 2 cm from the anal verge. One internal opening in the lower rectum was found with proctoscopy. The patient underwent fistulectomy. Microscopic examination showed SCC arising from the anal fistula, which was accompanied by vessel invasion. The tumor was observed to be continuous from the external opening but was not exposed to the internal opening of the rectal mucosa. Because human papillomavirus (HPV) infection was suspected, immunohistochemical analysis was performed, but showed no HPV infection. Two weeks after fistulectomy, abdominoperineal resection with lymph node dissection was performed. Histopathological examination revealed no remnant cancer tissue or lymph node metastasis. She was discharged after surgery without complications. Eight years after the operation, she complained of constant pain during micturition. Urological examination revealed urinary bladder cancer, and transurethral resection of the bladder tumor was performed. Histopathological examination revealed transitional cell carcinoma of the urinary bladder. Two years later, the patient died of metastatic urinary bladder cancer, without recurrence of the fistula cancer. Because the patients mother had died of urinary bladder cancer and she herself had metachronous urinary bladder cancer in addition to fistula cancer, we investigated whether microsatellite instability (MSI) and chromosomal instability correlated with fistula cancer development. Immunohistochemical analysis of formalin-fixed, paraffin-embedded surgical tumor specimens for p53, MLH1, and MSH2 was performed. The tumor specimens showed no MLH1 expression but did show normal MSH2 expression. p53 was not expressed. Five microsatellite loci were examined using the tumor specimens to detect MSI, namely two loci with mononucleotide runs (i.e., BAT25 and BAT26) and three loci with dinucleotide repeats (i.e., APC, Mfd15, and D2S123). The tumor specimens showed alternations in the repeated sequences of two loci (i.e., BAT26 and D2S123). As a result, the tumor was classified as MSI-H (high) according to the Bethesda criteria. Our patient had MSI and one of the smallest reported SCCs arising from recurrent anal fistulae.

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Onodi cells have been defined as posterior ethmoid cells that have pneumatized laterally and superiorly to the sphenoid sinus. They are often close to the optic nerve or the internal carotid artery. A patient complained of a sudden decrease in left eye visual acuity and left eye pain due to formation of a primary mucocele in an Onodi cell. A coronal computed tomography scan and magnetic resonance imaging were useful for diagnosing the mucocele.

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