A randomized clinical study on postoperative pain comparing the Polysoft patch to the modified Kugel patch for transinguinal preperitoneal inguinal hernia repair.

PURPOSE: The transinguinal preperitoneal approach is a relatively new technique for inguinal hernia repair. Two types of memory-ring mesh are available in Japan: the modified Kugel patch (MK) and the Polysoft patch (PP). We tested the hypothesis that the PP is noninferior to the MK with respect to chronic postoperative pain. METHODS: An unblinded randomized controlled trial was conducted to assess the noninferiority of PP compared to MK with a 5 % noninferiority margin. A total of 442 inguinal hernia patients operated on from November 2010 to December 2012 were included in this study. The primary endpoint was the pain score assessed by the visual analog scale (VAS) (0-1 vs. 2-10) 1 year after surgery. RESULTS: The patients were randomized to the PP and MK groups (n = 221 each). One year after surgery, 206 patients (95.4 %) in the PP group and 182 patients (89.6 %) in the MK group rated pain at 0-1 on the VAS scale. According to this rating, the PP group appeared not to be inferior to the MK group (95 % confidence interval, 0.7-10.7 %, P < 0.05). Furthermore, crude superiority tests, adjusting for 1 month of pain, denoted that the outcomes were significantly improved with the PP compared to the MK. CONCLUSIONS: The use of the PP was noninferior to the MK with respect to the severity of postoperative chronic pain scores 12 months after surgery.

Phase I clinical study of multiple epitope peptide vaccine combined with chemoradiation therapy in esophageal cancer patients.

BACKGROUND: Chemoradiation therapy (CRT) has been widely used for unresectable esophageal squamous cell carcinoma (ESCC) patients. However, many patients develop local recurrence after CRT. In this study, we hypothesized that the immunotherapy by peptide vaccine may be effective for the eradication of minimal residual cancer cells after CRT. This study was conducted as a phase I clinical trial of multiple-peptide vaccine therapy combined with CRT on patients with unresectable ESCC. PATIENTS AND METHODS: HLA-A*2402 positive 11 unresectable chemo-naïve ESCC patients were treated by HLA-A*2402-restricted multi-peptide vaccine combined with CRT. The peptide vaccine included the 5 peptides as follows; TTK protein kinase (TTK), up-regulated lung cancer 10 (URLC10), insulin-like growth factor-II mRNA binding protein 3 (KOC1), vascular endothelial growth factor receptor 1 (VEGFR1) and 2 (VEGFR2). CRT consisted of radiotherapy (60 Gy) with concurrent cisplatin (40 mg/m²) and 5-fluorouracil (400 mg/m²). Peptide vaccines mixed with incomplete Freund's adjuvant were injected subcutaneously once a week on at least 8 occasions combined with CRT. RESULTS: Vaccination with CRT therapy was well-tolerated, and no severe adverse effects were observed. In the case of grade 3 toxicities, leucopenia, neutropenia, anemia and thrombocutopenia occurred in 54.5%, 27.3%, 27.3% and 9.1% of patients, respectively. Grade 1 local skin reactions in the injection sites of vaccination were observed in 81.8% of patients. The expressions of HLA class I, URLC10, TTK, KOC1, VEGFR1 and VEGFR2 antigens were observed in the tumor tissues of all patients. All patients showed peptide-specific cytotoxic T lymphocytes responses in at least one of the 5 kinds of peptide antigens during the vaccination. Six cases of complete response (CR) and 5 cases of progressive disease (PD) were observed after the 8th vaccination. The 4 CR patients who continued the peptide vaccination experienced long consistent CR for 2.0, 2.9 4.5 and 4.6 years. CONCLUSIONS: A combination therapy of multi-peptide vaccine with CRT can successfully be performed with satisfactory levels of safety, and application of this combination therapy may be an effective treatment for patients with unresectable ESCC. TRIAL REGISTRATION: ClinicalTrial.gov, number NCT00632333.

Chronic pain and discomfort after inguinal hernia repair.

PURPOSE: The purpose of this study was to estimate the incidence and degree of persistent chronic pain after inguinal hernia repair performed in our hospitals. METHODS: We mailed a questionnaire on the frequency and intensity of postoperative inguinal pain and discomfort to 219 adult patients who had undergone inguinal hernia repair in one of our hospitals more than 3 months previously. RESULTS: There were 191 (87.2%) respondents, 28 (14.7%) of whom reported pain and 33 (17.3%) reported discomfort. The frequency of pain was "rare" in more than half of the patients who reported pain. No patient reported "continuous" pain. The intensity of the pain was "mild" in most of the patients, and none reported "pain that required a painkiller". The answers on the face scale questionnaire (score "0" to "10") were "0" or "1" for most of the patients. The incidence of chronic pain and/or discomfort was significantly higher in women than in men, and tended to be higher in patients who had undergone repair using onlay mesh. CONCLUSIONS: The frequency and intensity of persistent chronic pain or discomfort after inguinal hernia repair was not high or severe. These data will be useful for further studies to determine the best treatment for adult inguinal hernia.

Differentiation of femoral versus inguinal hernia: CT findings.

OBJECTIVE: The purpose of our study was to investigate the CT findings of femoral hernias, focusing on their differentiation from inguinal hernias. MATERIALS AND METHODS: We reviewed the records of 46 femoral hernias in seven centers (review of femoral hernias) and those of 215 groin hernias (femoral hernias, 11; inguinal hernias, 204) in one center (review of groin hernias). We evaluated the presence of hernia, extent of hernia sac based on the relationship between the hernia sac and the pubic tubercle (localized sac: sac was localized lateral to the pubic tubercle; or extended sac: sac extended medial to the pubic tubercle), and compression of the femoral vein on CT images. The chi-square test was used to assess the relationship between the CT findings and femoral versus inguinal hernias in the review of groin hernias. RESULTS: In the review of 46 femoral hernias, the lesions were detected on CT in 45. In the 45 lesions, all hernia sacs were localized, and 42 lesions showed venous compression. In the review of 215 groin hernias, all 11 femoral hernias had localized sacs with venous compression on CT. Of the 204 inguinal hernias, 98 lesions were detected on CT, 65 had extended sacs, and only 10 showed venous compression. Localized sacs with venous compression were seen much more often in the femoral hernias (11/11, 100%) than in the inguinal hernias (1/92, 1.1%) (p < 0.0001). CONCLUSION: CT images are useful to differentiate femoral hernias from inguinal hernias.

Usefulness and clinical significance of quantitative real-time RT-PCR to detect isolated tumor cells in the peripheral blood and tumor drainage blood of patients with colorectal cancer.

The clinical significance of isolated tumor cells (ITC) circulating in the blood of patients with colorectal cancer is unclear. In this study, we investigated the relationship between the presence of ITC that express carcinoembryonic antigen (CEA) and/or cytokeratin 20 (CK20) transcripts in the blood and the clinicopathological findings and prognosis using the quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay. We studied peripheral blood and tumor drainage blood from 167 patients with colorectal cancer. Quantitative real-time RT-PCR assay was able to detect one tumor cell in 3x10(6) peripheral blood mononuclear cells. Applying a cut-off value, CEA and/or CK20 (CEA/CK20) were detected in 10.2% (17/167) of the patients' preoperative peripheral blood samples and 34.1% (57/167) of the patients' tumor drainage blood samples. In the relationship between the CEA/CK20 of the blood and the clinicopathological factors, a significant correlation was demonstrated between the positivity of marker genes and the depth of invasion, venous invasion, lymph node metastasis, liver metastasis or stage. The disease-free and overall survival of patients with CEA/CK20-positive peripheral or tumor drainage blood was significantly shorter than that of marker gene-negative patients. CEA/CK20 transcripts in tumor drainage blood were independent factors for prognosis in disease-free survival and overall survival. These results suggest that detecting CEA/CK20 mRNA in tumor drainage blood by real-time RT-PCR has prognostic value in patients with colorectal cancer. Large scale and long-term clinical studies are needed to confirm the prognostic value of genetically detecting ITC in the peripheral blood.

Proposal of a new and simple staging system of colorectal liver metastasis.

AIM: To create a new, simple and useful staging system for colorectal liver metastasis analogous to the Tumor Node Metastasis classification system of International Union Against Cancer. METHODS: A retrospective review was undertaken of 81 consecutive patients who underwent partial hepatectomy for colorectal liver metastases (group 1). Clinical and pathological features of both primary and metastatic liver cancers were entered into a multivariate analysis to determine independent variables helpful in accurately predicting long-term prognosis after hepatectomy. Using selected variables, we created a new staging system like TNM classification. The usefulness of the new staging system was examined in a series of 92 patients from another hospital (group 2). RESULTS: Multivariate analysis showed that 81 patients in group 1 had significant multiple hepatic tumors with the largest tumor being more than 5 cm in diameter, resectable extrahepatic distant metastases, and independent prognostic factors for poor survival after hepatectomy. Using these three variables, we created a new staging system to classify patients with colorectal liver metastases. Finally, our new staging system classified the patients both in group 1 and in group 2. CONCLUSION: Our new staging system of colorectal liver metastasis is simple and useful for staging patients.

Proposal of criteria to select candidates with colorectal liver metastases for hepatic resection: comparison of our scoring system to the positive number of risk factors.

AIM: To select accurately good candidates of hepatic resection for colorectal liver metastasis. METHODS: Thirteen clinicopathological features, which were recognized only before or during surgery, were selected retrospectively in 81 consecutive patients in one hospital (Group I). These features were entered into a multivariate analysis to determine independent and significant variables affecting long-term prognosis after hepatectomy. Using selected variables, we created a scoring formula to classify patients with colorectal liver metastases to select good candidates for hepatic resection. The usefulness of the new scoring system was examined in a series of 92 patients from another hospital (Group II), comparing the number of selected variables. RESULTS: Among 81 patients of Group I, multivariate analysis, i.e. Cox regression analysis, showed that multiple tumors, the largest tumor greater than 5 cm in diameter, and resectable extrahepatic metastases were significant and independent prognostic factors for poor survival after hepatectomy (P < 0.05). In addition, these three factors: serosa invasion, local lymph node metastases of primary cancers, and post-operative disease free interval less than 1 year including synchronous hepatic metastasis, were not significant, however, they were selected by a stepwise method of Cox regression analysis (0.05 < P < 0.20). Using these six variables, we created a new scoring formula to classify patients with colorectal liver metastases. Finally, our new scoring system not only classified patients in Group I very well, but also that in Group II, according to long-term outcomes after hepatic resection. The positive number of these six variables also classified them well. CONCLUSION: Both, our new scoring system and the positive number of significant prognostic factors are useful to classify patients with colorectal liver metastases in the preoperative selection of good candidates for hepatic resection.

Phenotype-specific cells with proliferative potential are produced by polyethylene glycol-induced fusion of mouse embryonic stem cells with fetal cardiomyocytes.

Because cardiomyocytes lose the ability to divide upon differentiation, myocardial failure is assumed to be generally irreversible. For terminal cardiac insufficiency, the potential for regenerative treatment by stem cells, especially embryonic stem (ES) cells, offers hope for the future. Recent studies showed that stem cells fuse spontaneously with cells remaining in damaged tissues, and restore tissue function. To imitate spontaneous fusion in vivo, we used polyethylene glycol (PEG) in vitro to fuse mouse ES cells and fetal cardiomyocytes and analyzed the cytochemical properties of the fused cells. Confocal laser scanning microscopy coupled with lipophilic dye labeling of the living cell membranes showed that there were fused cells of ES cells and cardiomyocytes after PEG treatment. By flow cytometry, the fusion efficiency between ES cells and cardiomyocytes was estimated to be about 45% of the total resulting cells. When green fluorescent protein (GFP)-expressing ES cells were fused with cardiomyocytes, the fused cells had immunoreactivity for GFP in their cytoplasm and cardiac troponin I in their myofibrils. Some of these cells also expressed proliferating cell nuclear antigen up to 11 days after fusion, the last time point examined. This study shows that PEG-induced fusions of mouse ES cells and cardiomyocytes have the cardiomyocyte phenotype and proliferation potential.

Usefulness of immunomodulators for maturation of dendritic cells.

Biological response modifiers (BRMs) augment the cytotoxic activity of various effector cells by the induction of multiple cytokines and suppression of immunosuppressive factors. BRMs are used extensively in adjuvant therapy for gastric cancer in Japan. In dendritic cell (DC)-based vaccine therapy, the quality of DCs is important in inducing strong antitumor immunity. A good manufacturing practice (GMP) grade agent for DCs maturation is desirable for safety. Here we report the effects of two BRMs, OK432 and PSK, which are GMP grade agents for the functional maturation of DCs. OK432 and PSK were examined in vitro, and compared with lipopolysaccharide (LPS) and a cytokine cocktail (IL-1beta, TNF-alpha, IL-6 and PGE2). In the immunophenotypical analysis, the expression of CD80 and CD83 of DCs stimulated with OK-432 increased significantly compared with PSK and medium, and this up-regulation was the same as levels of DCs stimulated with cytokine cocktail. DCs stimulated with OK-432 showed significantly higher production of IL-12 and Th1-type cytokines (IL-2 and IFN-gamma) compared with DCs stimulated with LPS or cytokine cocktail. OK-432 stimulated DCs could induce the significantly high level of cytotoxic T cell activity compared with PSK-stimulated or unstimulated DCs. These results suggest that OK432 is a GMP-grade reagent that promotes functional maturation of DCs and could be applied in DC-based vaccinations.

Effective treatment with chemotherapy and surgery for advanced small cell carcinoma of the esophagus.

A 78-year-old man reported a persistent midthoracic pain, mild dysphagia, and an abdominal distention. An abdominal computed tomography scan showed massive ascites, extensive paracardial mass, a large mass which invaded the pancreas, and a mass of multiple para-aortic lymphadenopathies which involved the superior mesenteric artery. An upper gastrointestinal endoscopic study revealed an infiltrative, ulcerating tumor of the lower esophagus. Histological study of the biopsy specimens from esophageal tumor showed small cell carcinoma. After combination chemotherapy, an abdominal computed tomography scan showed a disappearance of asites, a partial response reduction of paragastric mass, peripancreatic mass and para-aortic lymphadenopathies. Histological study of the biopsy specimens from esophageal tumor showed a viable small cell carcinoma. In June 2001, the patient underwent lower esophagectomy and proximal gastrectomy combined with splenectomy and distal pancreatectomy through an abdominal approach. Histological findings of the resected specimen showed that the esophageal tumor was a small cell carcinoma which invaded into the submucosal layer, and both paracardial and peripancreatic tumors, and all lymph nodes had no cancer cells. The patient's postoperative recovery was uneventful and discharged without aggressive chemotherapy postoperatively. However, he eventually died of progression of the metastasis 21 months after first detection of the carcinoma. Patients with esophageal small cell carcinoma treated with surgery following chemotherapy and/or radiotherapy have been reported to survive longer than those treated with chemotherapy and/or radiotherapy. Therefore, surgical resection may be recommended as the second therapy that occasionally produces long-term remission and possibly long-term survival for patients with small cell carcinoma of the esophagus.

[Reduction of immunosuppression and shift to Th1 response by tumor-DC (dendritic cells) fusion vaccine].

Immunotherapy with tumor-dendritic cells (DC) fusion vaccine has been shown to induce an immune response against multiple tumor antigens including unknown tumor antigens. In this study, we examined immunological changes by tumor-DC fusion vaccine. Nine patients with advanced or recurrent gastrointestinal cancer that was unresponsive to standard surgical therapy and chemotherapy were enrolled in this study after informed consent was obtained. Monocytes were separated from the peripheral blood collected by leukapheresis, and were cultured with GM-CSF and IL-4 for 6 days. Then, TNF-alpha, IL-1beta, IL-6 and PGE2 were added for maturation of DC. Fusion of irradiated tumor cells and DC was created by polyethylene glycol and electroporation. The fusion vaccine was injected subcutaneously into the inguinal region four times every two weeks. There were no adverse effects or autoimmune responses after vaccination in any patients. The clinical response was stable in five patients, and disease progression in four patients. Delayed skin tests changed to positive in six of the nine patients after vaccination. IAP levels decreased in five patients and TGF-beta levels decreased in six patients. Th1/Th2 and Tc1/Tc2 balances improved in six of the nine patients after vaccination. In this study, it was shown that immunosuppressive factors, such as IAP and TGF-beta, and Th1 balance are useful as markers of immunomonitoring for tumor-DC fusion vaccine in patients with advanced or recurrent gastrointestinal patients.

[Perioperative nosocomial infection preventive measures].

For the prevention of perioperative nosocomial infection, 1) topical mupirocin treatment, 2) tight perioperative glycemic control and 3) immunonutrition are described. A large, prospective, randomized trial showed that the nasal application of mupirocin may effectively reduce postoperative Staphylococcus aureus nosocomial infection in the subgroup of patients who had S. aureus in their nares. Tight glycemic control after surgery, especially in the early period after operation, may also be effective in decreasing postoperative infection. In septic ICU patients, strict glycemic control even reduces ICU and hospital mortality rates. Several specific nutritional substrates such as arginine, omega-3 fatty acids, glutamine, and RNA have been shown to modulate host immune function. Some enteral formulas enriched with such immunonutrients have been commercially available in the USA and in Europe and are now available in Japan. Recent meta-analyses of randomized, controlled trials have shown that the administration of these formulas to elective surgical patients results in a significant reduction in the risk of developing infectious complications by approximately 50% and shortens the overall hospital stay.

[Molecular diagnosis of infectious disease].

With the development of molecular biology over the decades, molecular technologies have become available for clinical diagnosis of infectious disease. The principle of the technique is to detect specific genes of microorganisms. Compared with the traditional culturing techniques, molecular technologies have several advantages: 1) direct detection and rapid identification of organisms that are slow growing or those currently lacking a system for in vitro cultivation; 2) nucleic acid-based methods for epidemiologic typing of microorganisms; and 3) identification of genotypic markers of microbial resistance to specific antibiotics. When sufficient amounts of nucleic acid can be obtained, direct probe tests (simple molecular hybridization-based techniques) may be performed. In many instances with small amounts of sample nucleic acids, amplification techniques(e.g., polymerase chain reaction[PCR]) are extremely useful. However, the application of PCR to clinical specimens has potential pitfalls due to the susceptibility of PCR to inhibitors, contamination, and experimental conditions. In some cases, DNA fingerprinting or restriction fragment-length polymorphism analysis is used and it has become the method of choice for epidemiologic analysis. The major disadvantage of the molecular method is that we cannot differentiate whether the microorganisms identified are alive or dead and chemosensitivity testing cannot be performed.

Clinical significance of circulating tumor cells, including cancer stem-like cells, in peripheral blood for recurrence and prognosis in patients with Dukes' stage B and C colorectal cancer.

PURPOSE: Using multiple genetic markers, including cancer stem-like cells, we evaluated the clinical significance of circulating tumor cells (CTCs) as a prognostic factor for overall survival (OS) and disease-free survival (DFS) in the peripheral blood (PB) of patients with colorectal cancer (CRC) who had undergone curative surgery. PATIENTS AND METHODS: In a multi-institutional study, 735 patients with CRC were assigned to a retrospective training set (n = 420) or prospective validation set (n = 315). CTCs that expressed carcinoembryonic antigen (CEA), cytokeratin (CK) 19, CK20, and/or CD133 (CEA/CK/CD133) mRNA in PB were detected using real-time reverse transcription polymerase chain reaction assay. RESULTS: In the training sets, OS and DFS of patients who were positive for CEA/CK/CD133 were significantly worse than those of patients who were negative for these markers (P < .001). At each staging analysis, OS and DFS of patients with Dukes' stage B or C cancer who were positive for CEA/CK/CD133 were significantly worse than those of patients who were negative for these markers (P < .003 and P < .001 in Dukes' stage B; P < .001 in Dukes' stage C). In contrast, in patients with Dukes' stage A, no significant differences were seen between patients who were positive for these markers and those who were negative. Cox multivariate analysis demonstrated that CEA/CK/CD133 was a significant prognostic factor for OS (hazard ratio [HR], 3.84; 95% CI, 2.41 to 6.22; P < .001) and DFS (HR, 3.02; 95% CI, 1.83 to 5.00; P < .001). In particular, in patients with Dukes' stage B and C cancer, CEA/CK/CD133 demonstrated significant prognostic value. In validation sets, similar results were confirmed in patients with Dukes' stage B and C cancer. CONCLUSION: In patients with Dukes' stage B and C CRC who require adjuvant chemotherapy, detection of CEA/CK/CD133 mRNA in PB is a useful tool for determining which patients are at high risk for recurrence and poor prognosis.

Superior protective and therapeutic effects of IL-12 and IL-18 gene-transduced dendritic neuroblastoma fusion cells on liver metastasis of murine neuroblastoma.

Fusion vaccine of dendritic cells (DCs) and tumor cells has the advantage of inducing an immune response against multiple tumor Ags, including unknown tumor Ags. Using the liver metastasis model of C1300 neuroblastoma cells, we assessed the protective and therapeutic effects of fusion cells transduced with the IL-12 gene and/or the IL-18 gene. Improving the fusion method by combining polyethylene glycol and electroporation increased loading efficiency. In the A/J mice vaccinated with fusion cells modified with the LacZ gene (fusion/LacZ), IFN-gamma production and CTL activity increased significantly compared with that of DCs/LacZ, C1300/LacZ, or a mixture of the two (mixture/LacZ). With the transduction of IL-12 and IL-18 genes into the fusion cells (fusion/IL-12/IL-18), the level of IFN-gamma increased more than five times that of other fusion groups. In addition, NK cell activity and CTL activity increased significantly compared with that of mixture/LacZ, fusion/LacZ, DC/LacZ, or C1300/LacZ. In the protective and therapeutic studies of fusion cell vaccine, mice vaccinated with fusion/LacZ, fusion/IL-12, fusion/IL-18, or fusion/IL-12/IL-18 showed a significant decrease in liver metastasis and a significant increase in survival compared with mice given a mixture/LacZ, DCs/LacZ, or C1300/LacZ. In particular, the mice receiving fusion/IL-12/IL-18 vaccine showed a complete protective effect and the highest therapeutic effects. The present study investigates the improved loading efficiency of fusion cells and suggests that the introduction of IL-12 and IL-18 genes can induce extremely strong protective and therapeutic effects on liver metastasis of neuroblastoma.

Characterization of effector cells with anti-Candida activity obtained from murine bone marrow cells cultured in the presence of rhG-CSF: comparison between normal and CY-treated mice.

Bone marrow cells (BMC) obtained from normal and cyclophosphamide (CY)-treated mice were cultured in the presence of recombinant human granulocyte-colony stimulating factor (rhG-CSF) and their effector cell activities inhibiting growth of C. albicans were examined. When BMC from CY-treated mice were preincubated with 0.05 ng/ml of rhG-CSF, effector cells with enhanced anti-C. albicans activity were recovered in the adherent cell population, whereas anti-C. albicans activity of BMC from normal mice was found in the non-adherent cell population. During culture without the presence of rhG-CSF, nonadherent BMC, seemingly granulocytes, from normal mice showed apoptotic change, but addition of rhGCSF clearly inhibited this change. On the other hand, when BMC from CY-treated mice were cultured with rhG-CSF, adherent cells as the main effector had the appearance of monocytes. These differences between the effectors with anti-C. albicans activity obtained from normal and CY-treated mice are discussed.

A scoring system for the assessment of the risk of mortality after partial hepatectomy in patients with chronic liver dysfunction.

BACKGROUND/PURPOSE: The aim of this study is to evaluate a new scoring system, called the chronic liver dysfunction (CLD) score, for prediction of the surgical risk of partial hepatectomy in patients with chronic liver damage. Morbidity and mortality rates after gastroenterological surgery are high in patients with hepatic cirrhosis. Accordingly, it is very important to assess the surgical risk in such patients before surgery. Although the Child classification (or Child-Pugh score) has been a standard system, it did not always accurately predict patients at the risk of mortality after gastroenterological surgery, especially partial hepatectomy. METHODS: In 1985, we established a new system called the CLD score, reviewing the patients undergoing gastroenterological operations at one hospital. In the present study, we prospectively used the CLD score in 256 consecutive patients with chronic liver dysfunction who were treated surgically by partial hepatectomy, and investigated the usefulness of the CLD score concerning mortality. The results were compared with those of the Child-Pugh score (C-P score). RESULTS: After major hepatectomy, all the patients with CLD score exceeding 1.5 died of hepatic failure. After minor hepatectomy, all the patients with CLD score exceeding 2.5 died of hepatic failure. On the other hand, C-P score did not predict the outcome in these patients. CONCLUSIONS: Compared with the C-P score, which was considered the gold standard scoring system for assessing surgical risk for patients with chronic liver dysfunction, our CLD score provides a more reliable assessment of the risk of partial hepatectomy.