Assessment of decalcifying protocols for detection of specific RNA by non-radioactive in situ hybridization in calcified tissues.

For the best performance of in situ analysis of specific RNA expression in calcified tissues, it is necessary to choose an appropriate protocol to decalcify the tissues. We evaluated the usefulness of various acid-based decalcifying reagents with reference to 28 S rRNA staining by in situ hybridization using a thymine-thymine dimerized oligonucleotide probe. The reagents evaluated were 10% nitric acid, 10% HCl, 5% formic acid, 5% trichloroacetic acid, Morse's solution, Plank-Rychlo's solution, and K-CX solution, all of which are commonly used to decalcify tissues, and their effects on retention of morphology and RNA were compared with EDTA-based solutions. When normal mouse mandible was used as a model tissue, well-preserved morphology of ameloblasts was obtained from sections decalcified with Morse's solution, 10% HCl, Plank-Rychlo's solution, and K-CX solution, and best retention of 28 S rRNA was obtained with 5% formic acid and Morse's solution. We recommend Morse's solution to decalcify tissues to be processed for the rapid analysis of specific RNA expression. Indeed, we detected specific mRNAs strongly in sections treated with Morse's solution, and quantitative analysis showed that the ratio of signal intensities of 28 S rRNA and the specific mRNAs correlated with each other depending on decalcifying solutions.

[Intraoperative assessment of the ascending aorta by echogram during the CABG operation for the prevention of cerebral infarction].

While coronary artery bypass grafting operations (CABG), we started to use the echogram to evaluate the ascending aorta intraoperatively from December 1993. During a period from January, 1990, to June, 1995, 379 patients were treated with CABG. Intraoperative echogram was used in 127 cases (group I) and 252 cases unused (group II). The sites of the aorta for evaluation were following: site of cannulation, aortic clamping, and proximal anastomosis respectively. Depending on the echographical findings the operative procedures were changed as required. The incidence of postoperative cerebral embolism in group I was 0.8% (one patient) and in group II, 2.4% (6 patients). Although no significant difference was found in the cohort, rate of incidence in the group II was high. There were 4 cases who had significant atheromatous lesions clearly evaluated by echogram, which were not detected by palpation. We conclude that the intraoperative assessment of the ascending aorta by echogram is beneficial in prevention of cerebral infarction.

The two-fire, one-cartridge stapling method using a modified Endo-GIA.

The endoscopic stapler (Endo-GIA) was designed to divide tissue between two triple-stapled lines. The endoscopic surgeon frequently encounters situations where only stapling is required. Kirby described a staple closure method that uses a knifeless Endo-GIA cartridge. This method, although useful, has the problem of the modified Endo-GIA unit locking. Therefore, we devised a new technique for endoscopic stapling that involves two consecutive staplings using one cartridge without cutting. This method requires modification of the Endo-GIA system by removing the safety-lock system. In addition to enabling endoscopic stapling techniques without the risk of locking, the method can effect a significant cost savings.

[Preoperative assessment of aortic dissection by three-dimensional CT angiography].

In 21 patients with aortic dissection including 12 patients with acute aortic dissection, aortic lesions were evaluated preoperatively by newly developed three-dimensional CT angiography (3 D-CTA) using helical scan CT. Rapid and safe three-dimensional evaluation of aortic dissection was achieved with no complications. Three-dimensional structure of aortic dissection was well reflected in the image of 3D-CTA. Preoperative three-dimensional evaluation was useful to determine and proceed the operative procedure for the repair of aortic dissection. All patients were alive and results of surgical treatments were excellent. Intimal flaps and entries were visualized by multiple threshold display (MTD) method of 3D-CTA. Multiple threshold display method is an important three-dimensional reconstructive method and 3D-CTA may be a new useful and powerful diagnostic modality for aortic dissection.

Assessment of aortic disease using three-dimensional CT angiography.

The recently developed technique of three-dimensional CT angiography (3D-CTA) was applied to 68 patients with aortic disease. These patients were examined using a unique method of data collection: a helical scanner with continuous tube rotation and continuous table feed. The scanner was used in conjunction with administration of dynamic intravenous contrast material to enhance the vascular image. The group of patients included 35 with aortic dissection, 19 with true thoracic aneurysms, and 14 with abdominal aortic aneurysms. Three-dimensional evaluation was achieved in all patients with no complications. Surgical intervention was used in 45 patients with aortic dissections and aortic aneurysms, and 44 of these patients (98%) survived and were discharged. Preoperative 3D-CTA findings were quite similar to intraoperative findings, and were useful in determining operative procedures. Rapid and accurate assessment of aortic disease was achieved by 3D-CTA. Three-dimensional CT angiography can play an important role in the preoperative assessment of aortic dissections and aortic aneurysms leading to successful surgical treatment.

Assessment of tumor cell kinetics by monoclonal antibody Ki-67.

The expression of Ki-67 antigen in 71 patients with advanced gastric cancer was studied by immunohistochemical technique. Immunohistochemical staining with Ki-67 produced clear labeling of a portion of tumor cell nuclei, and the nucleoli stained intensely. The Ki-67 labeling rates of the 71 specimens ranged from 7.7 to 70.5% (mean: 29.2%; standard deviation: 12.9%). There was no significant association between Ki-67 labeling rates and macroscopic type, peritoneal metastasis, or serosal invasion. The tumors showing high Ki-67 labeling rates (greater than 25%) are more likely to have liver metastasis and lymph node involvement. Larger tumors, with a diameter greater than 6 cm, more frequently showed high Ki-67 labelling rate than those with a diameter less than 6 cm. When the Ki-67 labeling rate and 9 clinicopathologic parameters, as conventional prognostic factors, were entered simultaneously into the regression model, nodal status and Ki-67 labeling rate emerged as independent prognostic factors. These results indicate that the in situ determination of the growth fraction by Ki-67 antibody may be a reliable prognostic marker of advanced gastric cancer.

The value of transrectal ultrasonography in preoperative assessment for transurethral prostatectomy.

Transrectal ultrasonography was done in 19 healthy men and 236 patients with benign prostatic hyperplasia to elucidate the pathomorphology of the prostate and to determine preoperative prostatic size in patients who underwent transurethral prostatectomy. We found that the ultrasonographic examination allowed for an accurate estimation of the prostatic weight and a good assessment of the completeness of resection of the prostatic adenoma.