Safety and growth suppressive effect of intra-hepatic arterial injection of AdCMV-p53 combined with CDDP to rat liver metastatic tumors.

Surgical resection is thought to be the best treatment for liver carcinoma, including hepatocellular carcinoma and metastatic liver carcinoma if there are a small number of tumors. Liver carcinoma is one of the main causes of death from cancer worldwide. The prognosis of liver carcinoma is still poor. Mutation of p53, which is well known as a tumor suppressor gene, is observed in many cases of advanced liver carcinoma. Cancer gene therapy using p53, which transduces the wild-type p53 gene in the tumor, is a promising new strategy for treating liver carcinoma. Selective and less invasive gene delivery to the liver tumor is necessary for clinical liver tumor gene therapy. The first purpose of the current study was to determine the best way to deliver the gene of interest to the liver tumor selectively. The second purpose was to study the tumor suppressive effect of intrahepatic arterial injection of an adenovirus vector with the p53 gene (AdCMV-p53), followed by administration of CDDP and noting its side effects. We injected AdCMV-LacZ via hepatic arteries of rats bearing RCN-9 colon cancer metastasis in the liver. Injection via the hepatic artery resulted in more successful gene transduction to the liver tumor in a tumor-selective manner than did injection via the portal vein. At 48 hrs after arterial injection of AdCMV-p53, CDDP (3 mg/kg) was administered in the peritoneal cavity of each rat. The use of CDDP with arterial injection of AdCMV-p53 resulted in more extensive apoptosis in the rat liver tumors without any deterioration in liver function. In conclusion, hepatic arterial injection of an adenovirus vector is better than portal vein injection for gene transduction efficiency, and causes no liver function disorder even when the injection is combined with CDDP.

Identification of chemotherapeutic refractory cases based on human chorionic gonadotropin values among patients with low-risk persistent trophoblastic disease treated with 8-day methotrexate-folinic acid.

PURPOSE: The aim of the present study was to establish the accurate cutoff points of post-treatment serum beta-hCG values in identifying chemotherapeutic refractory cases among patients with low-risk persistent trophoblastic disease (PTD) treated with 8-day methotrexate-folinic acid as the primary therapy. MATERIALS AND METHODS: The values of serum beta-hCG measured before initiating treatment and weekly thereafter in 26 patients with low-risk PTD undergoing 8-day methotrexate-folinic acid treatment were analyzed. Thereafter, we determined the weekly cutoff points to identify the patient refractory for treatment by means of receiver-operating characteristic (ROC) plots analysis. RESULTS: The values of cutoff points in the pretreatment, the post-treatment 1st, 2nd, 3rd, and 4th week were 18.6, 15.0, 5.4, 3.4, and 2.0 ng/ml, respectively, and the value of accuracy during these weeks was appropriate (> 80%). When using the cutoff points of one and two weeks after initiating treatment, the accuracy in identifying chemotherapeutic refractory patients was 87.5% and 88.0%, respectively, with the highest values exceeding 85%. The sensitivity and specificity at one week were 92.9 and 80.0%, respectively. Similarly, the sensitivity and specificity at two weeks were 93.3 and 80.0%, respectively. CONCLUSION: These results suggest that the cutoff points of one and two weeks after initiating treatment are useful in identifying chemotherapeutic refractory patients among low-risk PTD patients, receiving 8-day methotrexate-folinic acid treatment.

Emergence of anti-inflammatory monocytes in long-term surviving hosts of IL-10-transduced liver allografts.

We previously reported that transduction of IL-10 to rat liver allografts facilitates survival prolongation after orthotopic liver transplantation (OLT). In the current study, we examined Lewis hosts of IL-10-transduced allografts that had survived longer than 50 days in order to characterize peripheral blood mononuclear cells (PBMC). Phenotype analysis of the PBMC demonstrated an 18-fold increase in monocytes (CD11b/c(+)) with a massive increase in the monocyte/lymphocyte ratio. The monocytes expressed downregulated MHC class II (RT1B) but upregulated Fcgamma receptors in comparison with monocytes from the control hosts. The capacity of enriched monocytes to secrete TNF-alpha in response to LPS stimulation was downregulated in the survivors, while production of IL-10 was comparable. The monocytes from long-term survivors significantly inhibited the donor antigen stimulated secretion of IFN-gamma and IL-2. Monocytosis characterized by a shift to anti-inflammatory monocytes is associated with survival prolongation in the hosts of IL-10 transduced liver allografts.

Egg maturation and parthenogenetic recovery of diploidy in the scorpion Liocheles australasiae (Fabricius) (Scorpions, ischnuridae).

In the scorpion Liocheles australasiae, egg maturation and parthenogenetic recoveries of chromosome number and nuclear DNA content were examined by histological, karyological observations and quantitative measurements of DNA. The primary oocyte becomes mature through two successive maturation divisions. The first maturation division takes place in the primary oocyte to produce a secondary oocyte and a first polar body. The second maturation division soon occurs in the secondary oocyte, in which the nucleus is divided into a mature egg nucleus and a second polar body nucleus, not followed by cytoplasmic fission. The first polar body, in one case, was successively divided into two second polar bodies; in the other case it was not divided. In either case, these polar bodies remained attached to the early embryo. The fate of these polar bodies during further embryogenesis were studied. In the karyological analysis, the chromosome number was divided into two groups, one from 27-32, the other was 54-64. The former was presumably the metaphase chromosome number at the meiotic division; the latter was presumably the metaphase chromosome number at the mitotic division. DNA content in the diploid nucleus of the primary oocyte, doubled before the maturation divisions, was reduced through the maturation divisions by one-half in the nuclei of the secondary oocyte and the first polar body and by one-fourth in the nuclei of the egg and the second polar bodies. The first reduction of DNA content corresponded to halving the number of the chromosomes in the first maturation division and the second to the nuclear division in the secondary oocyte. These reductions represent a common process of egg maturation, except the final production of the mature egg with two haploid nuclei, an egg nucleus, and a second polar body nucleus. These two nuclei, which were formed apart in the mature egg, drew near to fuse into a zygote nucleus. The chromosome number and nuclear DNA content were doubled in the zygote and each blastomere in embryos, supporting the hypothesis that the egg nucleus fuses with the second polar body nucleus and this conjugation initiates subsequent embryonic development.

Prolongation of liver allograft survival after interleukin-10 gene transduction 24-48 hours before donation.

BACKGROUND: Interleukin- (IL) 10 may be a potent regulator for controlling of allograft rejection. A single administration of IL-10 is not effective for controlling graft rejection. Gene transfer is an attractive vehicle for prolonging the expression of short-lived proteins. METHODS: Donor or recipient livers were transduced with 1 x 10(10) p.f.u. of replication-deficient adenovirus vectors harboring human IL-10 cDNA (AdCMVhIL-10) via the ileocecal vein before or after rat orthotopic liver transplantation. RESULTS: DA allografts given AdCMVhIL-10 24-48 hr before donation survived for more than 56 days in Lewis recipients, although DA allografts given the adenovirus vector 7 days or 6 hr before, and 3 days after transplantation were rejected within 30 days in recipients. Serum levels of human IL-10 in gene-transferred rats were maximum from day 2 to 7. The serum level of human IL-10 then decreased gradually, and human IL-10 was not detected by ELISA 30 days after gene-transduction. In gene-transduced long-term surviving liver allografts, IL-10 was expressed, and the expression of IL-4 was also up-regulated on posttransplant day 3, despite the expression of Th1 cytokines (IL-2 and interferon-gamma), although in rejected liver allografts, IL-2 and interferon-gamma were expressed without expression of IL-4 and IL-10. CONCLUSIONS: The prolongation of survival of IL-10 cDNA transferred liver allografts might be due to inhibition of the early phase of alloimmune-response by over expression of IL-10, despite the expression of IL-2 and interferon-gamma.

Study on eight patients with malignant tumors after renal transplantation.

The high incidence of cancer after renal transplantation is now a critical concern since the graft survival rate has been improved extensively. We experienced 9 malignancies in 8 patients out of 168 recipients up to December 31, 1999 in our hospital, consisting of a case of gastric plasmacytoma and cases of cancer in the liver (2), thyroid (2), prostate (1), breast (1), sigmoid colon (1) and gall-bladder (1). Two patients were diagnosed as having tumors within 3 months after transplantation, suggesting post-transplant acceleration of growth of the latent tumors. The other patients were diagnosed at an average of 128 months, ranging from 84 to 263 months after transplant. Two patients died of gastro-intestinal bleeding and acute heart failure. Four patients died directly of progressive neoplasm within 3 months after diagnosis. These results suggest that the course of malignancies developing in post-transplant recipients is more aggressive than that expected in non-transplant patients, and it is very important to intensively follow long-term surviving cases to detect the malignant tumors as early as possible.

Laparoscopic cholecystectomy in a patient on continuous ambulatory peritoneal dialysis.

The patient was a 72-year-old man who was receiving continuous ambulatory peritoneal dialysis (CAPD) with a diagnosis of chronic renal failure. Although his response to dialysis therapy was favorable, right hypochondralgia and fever occurred, and gallstones were detected by abdominal ultrasonography and computed tomography. Drip-infusion cholangiography (DIC) revealed neither dilation nor calculus in the common bile duct. The patient was diagnosed as having acute cholecystitis and cholecystolithiasis and, in consideration of his general condition, laparoscopic cholecystectomy was carried out. Pneumoperitoneum was performed through a CAPD tube, and a 10 mm-trocar was carefully introduced through a supraumbilical incision so as not to injure the CAPD tube. Since intraoperative cholangiography showed a condition similar to preoperative DIC, only cholecystectomy was undertaken. The postoperative course was uneventful, with neither postoperative hemorrhage nor leakage of dialysate from the wound.

Comparison of the anterior colporrhaphy procedure and the Marshall-Marchetti-Krantz operation in the treatment of stress urinary incontinence among women.

OBJECTIVE: To compare retrospectively the efficacy of the anterior colporrhaphy procedure (AC) and the Marshall-Marchetti-Krantz operation (MMK) in the treatment of stress urinary incontinence (SUI). METHODS: A retrospective analysis through a review of the medical records of Japanese women with stress urinary incontinence who were surgically treated at Kyushu University Hospital from 1980 through 1996. A questionnaire regarding the current status of urinary incontinence was sent to all patients. RESULTS: A total of 103 patients could be evaluated, 77 of whom had undergone an AC, and 26 of whom had undergone an MMK. Postoperative complications were more common in the AC group (p < 0.05). There were no significant differences between the 2 groups in terms of the duration of hospital stay or postoperative catheterization. The long-term subjective cure rates of the ACs and the MMKs were 55% and 58%, respectively. CONCLUSIONS: The AC and the MMK were equally effective in treating stress urinary incontinence, and both showed decreased long-term subjective cure rates. The recurrence rate did not differ between the AC and the MMK. The AC had more postoperative complications and shorter recurrence intervals.

Allograft transduction of IL-10 prolongs survival following orthotopic liver transplantation.

Interleukin-10 (IL-10) is an ideal candidate cytokine for suppressing the alloimmune response in transplantation. To determine whether genetic modulation of the hepatic graft with IL-10 could prolong survival following orthotopic liver transplantation, we constructed a replication-deficient adenovirus vector expressing human IL-10 (AdCMVhIL-10). Intraportal injection of this vector into a donor rat 24-48 h before grafting resulted in efficient release of IL-10 into the circulation of a recipient rat after transplantation. Moreover, levels of hIL-10 from the suprahepatic vena cava were significantly (1.48-fold) higher than those from the infrahepatic vena cava (P = 0.013), indicating local IL-10 production within the transduced hepatic graft. AdCMVhIL-10 induced a prolongation of median survival to more than 87 days, with two of five transduced grafts showing more than 100 days of ongoing survival, when compared with 11 days for grafts transduced with a control adenovirus vector carrying the E. coli beta-galactosidase gene (P = 0.0021) and 11 days for untreated grafts (P = 0.0021). Pathological findings occurring in the AdCMVhIL-10-transduced hepatic grafts revealed no evidence of progressive rejection reaction resulting in graft failure. These results demonstrate that hepatic grafts modulated by IL-10 gene transfer make local and effective immunosuppression feasible in the transplantation setting.

Is nuclear expression of Y box-binding protein-1 a new prognostic factor in ovarian serous adenocarcinoma?

BACKGROUND: Nuclear expression of Y box-binding protein (YB-1), a member of the DNA binding protein family, has been reported to be much more highly concentrated in cisplatin-resistant cell lines than in their parental counterparts, suggesting an ability to limit cisplatin sensitivity. Moreover, YB-1 plays a key role in P-glycoprotein expression. Because ovarian carcinoma traditionally has been treated with cisplatin-based chemotherapy, the sensitivity of the tumors to chemotherapy could reflect a particular prognosis in patients with ovarian carcinoma. The aim of the current study was to determine whether YB-1 expression correlated with prognosis in ovarian serous adenocarcinoma patients. METHODS: The expression of YB-1 in the nucleus was examined immunohistochemically in 42 paraffin embedded primary Stage III (International Federation of Gynecology and Obstetrics) serous ovarian carcinoma tumors extirpated by primary surgery at Kyushu University Hospital between 1985-1995. RESULTS: Of the 40 primary ovarian tumors examined, 12 (30%) were positive for YB-1 expression in the nucleus. There was no significant difference in intraperitoneal stage, histologic grade, or residual tumor size after primary surgery between patients with tumors with positive and those with negative nuclear expression of YB-1 protein. The disease free survival curve for patients whose tumors were positive for nuclear expression of YB-1 protein was significantly worse than that for patients whose tumors were negative (P = 0.0025). P-glycoprotein was overexpressed in 4 of 12 tumors with nuclear YB-1 expression (33%) but there was no statistical significance between the expression of nuclear YB-1 and P-glycoprotein. CONCLUSIONS: The expression of YB- 1 protein in the nucleus may be considered a useful prognostic marker and also may reflect the sensitivity of ovarian serous adenocarcinoma to chemotherapy.

Granular cell tumor of the breast: report of a case.

A case of granular cell tumor of the breast in a 43-year-old woman is described. The patient presented with a painless mass in the upper-outer quadrant of her right breast. Mammography showed a spiculated tumor and ultrasonography demonstrated a hypoechoic mass with an irregular border. Magnetic resonance (MR) mammography revealed a homogeneous enhanced mass in T1-weighted images using Gd-DTPA and a ringed high-intensity area around the mass in T2-weighted images. Fine-needle aspiration cytology failed to show any malignant cells. A partial resection of the breast was performed and histological examination revealed a granular cell tumor. Granular cell tumors are generally always benign, but they may be misdiagnosed as malignant tumors because of their mammographic and ultrasonographic findings. MR mammography did not reveal a typical breast cancer in either T1- or T2-weighted images in the present case. This case illustrates the need for care in preoperative examinations in order to avoid overdiagnosis of breast cancer.

Predicting pelvic lymph node metastasis in endometrial carcinoma.

BACKGROUND: To determine the possibility of individualizing the pelvic lymph node dissection in patients with endometrial cancer, the relationship between pelvic lymph node (PLN) metastasis and various prognostic factors was retrospectively investigated. METHODS: From 1979 to 1994, 175 patients with endometrial carcinoma were treated with either total or radical hysterectomy combined with a PLN dissection as initial therapy. The prognostic factors examined included clinical stage, patient age, histological grade, the microscopic degree of myometrial invasion (DMI), cervical invasion, adnexal metastasis, and macroscopic tumor diameter (TD). RESULTS: Of the 175 patients undergoing PLN dissection, 24 (14%) had PLN metastasis. An endometrial cancer with PLN metastasis had a significantly longer diameter than those without PLN metastasis. The frequency of PLN metastasis increased along with increases in tumor diameter. A logistic regression analysis revealed DMI and TD to be independently correlated with PLN metastasis. The formula based on the coefficients of TD and DMI obtained from the analysis also showed a good correlation, which allowed us to estimate the probability of patients having PLN metastasis. CONCLUSIONS: DMI and TD could accurately estimate the status of PLN in endometrial carcinoma patients.

Factors affecting postoperative prognosis in the solitary-nodule type of hepatocellular carcinoma: experience of 132 cases in our institute.

A retrospective analysis of clinical and pathological factors was performed on 132 surgical cases with solitary-nodule type HCC in our hospital. The overall cancer-free survival rates after 1, 3 and 5 years were 82.2%, 42.3% and 26.5%, respectively. With univariate analysis, the significant prognostic factors for survival were tumor size, cancer cell infiltration of the fibrous capsule of the tumor (fc-inf), invasion into portal vein (vp), and intrahepatic metastasis (im), while significant prognostic factors for non-recurrence were tumor size, fc-inf, vp, im, Edmondson-Steiner's classification and perioperative blood transfusion. With multivariate analysis for recurrence, significant factors were vp, clinical stage (CS), and perioperative blood transfusion. Therefore, prognostic factors for long-term survival in surgical cases of HCC are thought to be good hepatic function, absence of portal invasion, and avoidance of perioperative blood transfusion if possible.

[The mechanism of donor specific unresponsiveness in renal transplant recipients].

A study was conducted to clarify the mechanism of donor specific immunological unresponsiveness in renal transplant recipients with a well functioning kidney, since this might provide a clue to the nature of donor specific immunosuppression. Peripheral blood lymphocytes (PBL) were obtained from three renal transplant recipients, the donors (mother), the fathers and healthy volunteers as 3rd party and used in the present study. PBL from one of the three renal transplant recipients, which were donor specific unresponsive in MLR (Mixed Lymphocyte Reaction) and CML (Cell Mediated Lympholysis), suppressed the MLR, CML and synthesis of IL-2 from 3rd party PBL in a double chamber assay only when stimulated by the donor PBL. This suppression was abolished by the treatment with CD3 or CD8 antibodies and complement. RT-PCR was performed to investigate the suppressive effect of the recipient's PBL on IL-2 mRNA expression. In the double chamber MLR, expression of IL-2 mRNA by the PBL from 3rd party was also suppressed only when the recipient's PBL were stimulated by the donor PBL. These results indicate that one of the mechanisms responsible for donor specific immunological unresponsiveness is the presence of donor specific suppressor T cells in recipient's PBL and that these cells project suppressive effect on lymphocytes by producing a humoral suppressive factor(s) that suppress the expression of the IL-2 mRNA only when stimulated by donor PBL.