Trabecular structure and composition analysis of human autogenous bone donor sites using micro-computed tomography.

OBJECTIVES: To evaluate the bone microstructure of autogenous graft bone in elderly people (mean age, 66 years), we compared the bone volume/total volume and bone mineral density of four donor sites that are commonly harvested for maxillofacial surgery and dental implant treatments, using X-ray micro-computed tomography. METHODS: Eighteen Japanese cadavers were included in this study. Overall, 66 harvested bones (mandibular symphysis, mandibular ramus, ilium, and tibia) were studied. Micro-computed tomography scans of four sites were performed to analyze the trabecular structures, bone mineral density, and bone volume/total volume in these bones. RESULTS: The mandibular symphysis bones showed the highest bone volume/total volume and bone mineral density at the four sites. There was a significant difference in the bone volume/total volume between the mandibular symphysis and tibia groups. There was also a significant difference in bone mineral density between the mandibular symphysis group and the ilium and tibia groups. In the three-dimensional observations, the structures of the mandibular trabecular were plate-type. The structures of the tibial bone were mixtures of plate- and rod-types. In the ilium, most trabecula were rod-shaped. CONCLUSIONS: Mandibular symphysis and ramus had a higher bone volume/total volume and bone mineral density of the four sites and did not show regressive changes in our findings. Mandibular bone is the most suitable source of autogenous graft bone material because of its superior bone quality and quantity.

Portal vein resection using the no-touch technique with a hepatectomy for hilar cholangiocarcinoma.

OBJECTIVES: To assess the safety and feasibility and discuss the oncological impact of a portal vein resection using the no-touch technique with a hepatectomy for locally advanced hilar cholangiocarcinoma. PATIENTS AND METHODS: From 2005 to March 2009, 49 patients with hilar cholangiocarcinoma underwent a major right-sided hepatectomy with curative intent. Portal vein resection was performed using the no-touch technique in 36 patients (PVR group) but the portal vein was not resected in the other 13 patients (NR group). Peri-operative data and histological findings were compared between the two groups. Moreover, tumour recurrence and survival rates after surgery were calculated and compared for each group. RESULTS: Although the tumours of the patients in the PVR group were more locally advanced, the residual tumour status and tumour recurrence rate were similar and there was no significant difference in long-term survival between the two groups: 5-year survival rates in the PVR and NR groups were 59% and 51%, respectively (P = 0.353). In-hospital mortality was encountered in 2 of the 49 patients. CONCLUSION: A portal vein resection using the no-touch technique with a right-sided hepatectomy had a positive impact on survival and is feasible in terms of long-term outcomes with acceptable mortality.

A multicentre randomised trial to compare the efficacy of omeprazole versus rabeprazole in early symptom relief in patients with reflux esophagitis.

BACKGROUND: Proton pump inhibitors (PPIs) are affected by cytochrome P450 2C19 (CYP2C19) polymorphisms. This study compared the effect of two PPIs on early symptom relief in Japanese patients with reflux esophagitis, classified by the CYP2C19 phenotype. METHODS: Patients with reflux esophagitis were randomised to treatment with omeprazole 20 mg or rabeprazole 10 mg once daily. The CYP2C19 phenotype [homozygous extensive metaboliser (homoEM), heterozygous extensive metaboliser (heteroEM) or poor metaboliser (PM)] of each patient was determined. The primary efficacy endpoint was early, sufficient (Global Overall Symptom scale score 1 or 2), sustained (maintained for ≥7 days) reflux symptom relief. RESULTS: Of the 199 patients included in this analysis, the proportion achieving sufficient, sustained reflux symptom relief was higher with omeprazole than with rabeprazole on day 1 (35.6 vs. 22.4%; p = 0.041) and day 2 (43.6 vs. 28.6%; p = 0.028); there was no significant difference between the two groups on days 3-7. Among patients with the CYP2C19 PM phenotype, sufficient, sustained reflux symptom relief was higher with omeprazole than with rabeprazole on days 4-7 (62.5-66.9 vs. 31.6%; p ≤ 0.03); differences were not significant on days 1-3, or among those with the homoEM or heteroEM phenotypes on days 1-7. CONCLUSIONS: In Japanese patients with reflux esophagitis, omeprazole 20 mg is more effective than rabeprazole 10 mg at achieving early, sufficient, sustained reflux symptom relief in individuals with the CYP2C19 PM phenotype, and is similarly effective to rabeprazole 10 mg in those with heteroEM or homoEM phenotypes.

[Gastric ulcer and duodenal ulcer].

Peptic ulcer diseases are improved by strong acid suppression therapy. However, peptic ulcer diseases are often relapsed after discontinuing of acid suppression therapy. After discovery of Helicobacter pylori (H. pylori) in 1984, H. pylori is known as the strongest causal agent for peptic ulcer diseases. In November 2000, eradication therapy for H. pylori infection was approved under the Japanese system of health insurance. H. pylori eradication reduces ulcer recurrence dramatically. Recently, peptic ulcer diseases induced by NSAIDs and/or aspirin are increasing. In prevention of NSAIDs and/or aspirin induced peptic ulcer recurrence, H. pylori eradication plus PPI treatment is recommended. Several reports described occurrence of newly-developed gastric cancer after H. pylori eradication. Careful follow-up upper gastrointestinal endoscopy is necessary after H. pylori eradication.

Modified duodenum-preserving pancreas head resection for low-grade malignant lesion in the pancreatic head.

BACKGROUND/OBJECTIVES: Our institution has utilized a duodenum-preserving pancreas head resection (DPPHR) procedure for management of low-grade malignant lesions within the head of the pancreas, but this has resulted in a higher rate of postoperative complications, including pancreatic fistula and ischemic bile duct injury. To avoid these complications we recently modified DPPHR to resect all the parenchyma around the pancreatic head and to preserve the epicholedochal plexus around the bile duct. The goal of this study was to investigate outcomes with postoperative complications and disease control following this modified procedure. METHODS: Twenty-one consecutive patients underwent DPPHR between 1994 and 2011. Patients were retrospectively classified into one of two groups: the conventional DPPHR group (cDPPHR) or the modified DPPHR group (mDPPHR). Perioperative factors and postoperative complications were compared between these two groups. RESULTS: The median age of the 21 patients was 61 (23-77) years, and the median follow-up period was 51 months. Intra-operational blood loss was significantly smaller and duration of hospital stay was significantly shorter in the mDPPHR group than in the cDPPHR group, respectively. The rate of pancreatic fistula was markedly lower in the mDPPHR group (2/13; 15%) than in the cDPPHR group (7/8; 88%) (P = 0.0022). For neoplastic lesions, the surgical margin was negative in all cases, and local recurrence has not occurred in either group. CONCLUSIONS: For selected patients, modified DPPHR may provide clinical benefits in terms of less complications associated with shorter hospital stay.

Method for the validation of immunohistochemical staining using SCID mouse xenografts: expression of CD40 and CD154 in human non-small cell lung cancer.

This report proposes a concept for the standardization of immunohistochemical evaluation. Immunohistochemical staining has several problems associated with the sensitivity of the technical process and standardization of the assessment of potent staining. We provided data focusing on this concept through immunostaining for CD154 in non-small cell lung cancer (NSCLC). We used two types of anti-CD154 antibody as primary antibodies in immunohistochemical staining, as previously reported. Western blot analysis confirmed strong CD154 expression in the cultured cell line PC10, but not in LK2. We also assessed CD154 expression in SCID mouse xenografts of these cell lines. SCID xenograft data on western blot analysis were consistent with those of cultured cell lines. These xenografts could thus be used as positive or negative tissue controls for CD154 immunostaining. Primary antibodies should therefore be confirmed as recognizing target lesions, while control tissue specimens should be objectively confirmed as having target products using another experimental method. Our method would allow results to be unified at more than one laboratory and could act as an objective control assessment method in immunohistochemistry.

Clinical evaluation of the feasibility of minimally invasive surgery in esophageal cancer.

INTRODUCTION: Open thoracotomy laparotomy with extended dissection for esophageal cancer is associated with problems such as delayed postoperative recovery and decreased quality of life. In contrast, in minimally invasive surgery, these problems can be improved. In the present study, we investigated the feasibility of minimally invasive surgery in esophageal cancer. METHODS: In this retrospective study, we evaluated esophagectomy performed by the same surgeon in 98 patients with thoracic esophageal cancer. Open surgery was performed in 30 patients (open group), and minimally invasive surgery was performed in 68 patients (MIS group). We compared the invasiveness and radical cure of cancer by minimally invasive surgery with that of open surgery. RESULTS: Comparison between the open and MIS groups showed that intraoperative blood loss, intraoperative and postoperative transfused blood volume, and surgical site infection rates were significantly lower in the MIS group. The duration of postoperative endotracheal intubation and hospital stay were significantly shorter in the MIS group. The histopathologic type was squamous cell carcinoma in 93.3% in the open group and 92.6% in the MIS group. The respective 3-year survival rates were 36.7% and 71.5%, and the respective 5-year survival rates were 26.7% and 61.5%. CONCLUSION: Based on a historical control study at a single institution, we are unable to conclude that minimally invasive surgery is superior to open surgery. However, our results indicate that minimally invasive surgery is feasible as a surgical procedure in esophageal cancer.

Evaluation of extra capsular lymph node involvement in patients with extra-hepatic bile duct cancer.

BACKGROUND: Lymph node metastasis is one of the most important prognostic factors for extra-hepatic bile duct carcinoma (ExHBDC). Extra capsular lymph node involvement (ExCLNI) is the extension of cancer cells through the nodal capsule into the perinodal fatty tissue. The prognostic impact of ExCLNI has been shown to be significant mainly in head and neck malignancies. Recently, the prognostic impacts of ExCLNI have evaluated in gastrointestinal malignancies. However no data is available regarding the incidence and prognostic significance of extra-capsular lymph node involvement (ExCLNI) in resectable ExHBDCs. The aim of the present study is first to evaluate the incidence of ExCLNI in surgically-treated ExHBDCs and second, to determine the prognostic impact of ExCLNI in patients with surgically-treated ExHBDCs. METHODS: A total of 228 patients (110 cases of hilar cholangiocarcinoma and 118 cases of distal cholangiocarcinoma) with surgically-treated ExHBDCs were included in this retrospective study. ExCLNI was defined as the extension of cancer cells through the nodal capsule into the perinodal fatty tissue. The existence of ExCLNI and its prognostic value were analyzed as a subgroup of lymph node metastasis. RESULTS: ExCLNI was detected in only 22% of patients with lymph node metastasis of surgically-treated ExHBDC. The presence of ExCLNI correlated with distal cholangiocarcinoma (p = 0.002). On univariate analysis for survival, perineural invasion, vascular invasion, histological grade, and lymph node metastasis were statistically significant factors. On multivariate analysis, only lymph node metastasis was identified as a significant independent prognostic factor in patients with resectable ExHBDC. Subgroups of lymph node metastasis including the presence of ExCLNI, location of lymph node metastasis, and the number of lymph node metastasis had no statistically significant impact on survival. CONCLUSION: ExCLNI was present in only 22% of the LNM (7% of overall patients) in patients with surgical treated ExHBDCs. And ExCLNI would have no impact on the survival of patients with surgically-treated ExHBDCs.

The immunological impact of neoadjuvant chemotherapy on the tumor microenvironment of esophageal squamous cell carcinoma.

BACKGROUND: Esophageal cancer is an aggressive cancer with poor prognosis. However, little is known about the immune response in the tumor microenvironment after neoadjuvant chemotherapy. PURPOSE: To investigate the immunological impact of neoadjuvant chemotherapy in the tumor microenvironment of esophageal squamous cell carcinoma. METHODS: Eighteen patients with esophageal squamous cell carcinoma with and without neoadjuvant chemotherapy were analyzed using immunohistochemical methods for human leukocyte antigen (HLA) class I heavy chain, CD4-, CD8-, and Foxp3-positive cell infiltration. RESULTS: The number of CD4 T cells in the stroma and within the cancer nest was significantly higher in the neoadjuvant chemotherapy group. The number of CD8 T cells in the stroma was significantly higher in the neoadjuvant chemotherapy group. HLA class I expression was more downregulated in the control group compared with the neoadjuvant chemotherapy group. CONCLUSION: Neoadjuvant chemotherapy utilizing 5-fluorouracil and cisplatin in esophageal squamous cell carcinoma is useful to induce CD4 and CD8 T lymphocytes in the tumor microenvironment and to maintain HLA class I expression levels in combination with its direct cytotoxic effects.

Continuous proton pump inhibitor treatment decreases upper gastrointestinal bleeding and related death in rural area in Japan.

BACKGROUND AND AIMS: Proton pump inhibitors (PPI) have been rarely used for prevention of upper gastrointestinal bleeding (UGIB) induced by non-steroidal anti-inflammatory drugs (NSAIDs) and/or aspirin in Japan. The increased incidence of UGIB in the aged society is becoming a serious problem. The aim of this study was to retrospectively evaluate whether PPI can prevent UGIB. METHODS: We examined records of 2367 patients (aged 67.9 ± 15.1 years, male 1271) attending the only hospital serving the rural area, with little population movement. We investigated the correlation between the frequency of usage of medicine (PPI, histamine 2 receptor antagonists [H2RA], NSAIDs, aspirin) and incidence of UGIB over 12 years. UGIB was defined as cases with hematemesis and/or melena and definite bleeding at upper gastrointestinal endoscopy. The annual incidence of UGIB of inhabitants (16,065 ± 375.3 persons/year) was evaluated. The frequency of usage of medicine was compared with the total number of patients prescribed any medication (1080 ± 33.2 persons/year). RESULTS: The frequency of PPI usage has increased significantly 4.6%→30.8% (P < 0.05). NSAIDs and aspirin usage increased significantly in the latter half of the survey period (P < 0.05). The annual incidence of UGIB significantly decreased 160.8 →23.6/100,000 inhabitants per annum (P ≤ 0.05) due to widespread use of PPI. No patients died due to UGIB after 2006. The incidence of UGIB and the prevalence of PPI usage were found to have a negative correlation (r = -0.804, P = 0.0016). CONCLUSIONS: By widespread use of PPI, UGIB and related death has declined significantly. This survey showed that continuous PPI treatment decreases UGIB and related death in community medicine.

Down-regulation of Human Leukocyte Antigen class I heavy chain in tumors is associated with a poor prognosis in advanced esophageal cancer patients.

The HLA class I antigen processing machinery (APM) plays a crucial role in the anticancer immune response. The aim of this study was to assess the clinical significance of APM components in esophageal cancer. A total of 11 esophageal cancer cell lines were evaluated by Western blot analysis for 13 HLA class I APM components. There was a different expression pattern among cancer cell lines for HLA class I heavy chain (HLA-HC), ß2 microglobulin, Tapasin, TAP-1, TAP-2, LMP-7 and LMP-10. Immunohistochemical staining utilizing a tissue microarray method for HLA class I APM expression showing different expression patterns among cell lines was performed for 95 surgical specimens from patients with esophageal cancer. Prognostic factors were the down-regulation of HLA-HC, and the up-regulation of ß2 microglobulin and TAP-1 in the cancer tissues. Multivariate analysis using a Cox regression model indicated that the down-regulation of HLA-HC, and up-regulation of TAP-1 in cancer tissues are independent, unfavorable prognostic factors (hazard ratio, 2.361 and 2.297; P=0.0141 and 0.0145, respectively). Although there was no significant difference in survival for selected p-stage I and II patients (n=54) in all APM components, only down-regulation of HLA-HC was an unfavorable prognostic factor by a Cox regression model for selected p-stage III and IV patients (n=41). In conclusion, the current results suggest that the down-regulation of HLA-HC in tumors is especially associated with a poor prognosis among advanced esophageal cancer patients.

Integrated care through disease-oriented clinical care pathways: experience from Japan's regional health planning initiatives.

INTRODUCTION: In April 2008, Japan launched a radical reform in regional health planning that emphasized the development of disease-oriented clinical care pathways. These 'inter-provider critical paths' have sought to ensure effective integration of various providers ranging among primary care practitioners, acute care hospitals, rehabilitation hospitals, long-term care facilities and home care. DESCRIPTION OF POLICY PRACTICE: All 47 prefectures in Japan developed their Regional Health Plans pursuant to the guideline requiring that these should include at least four diseases: diabetes, acute myocardial infarction, cerebrovascular accident and cancer. To illustrate the care pathways developed, this paper describes the guideline referring to strokes and provides examples of the new Regional Health Plans as well as examples of disease-oriented inter-provider clinical paths. In particular, the paper examines the development of information sharing through electronic health records (EHR) to enhance effective integration among providers is discussed. DISCUSSION AND CONCLUSION: Japan's reform in 2008 is unique in that the concept of 'disease-oriented regional inter-provider critical paths' was adopted as a national policy and all 47 prefectures developed their Regional Health Plans simultaneously. How much the new regional health planning policy has improved the quality and outcome of care remains to be seen and will be evaluated in 2013 after the five-year planned period of implementation has concluded. Whilst electronic health records appear to be a useful tool in supporting care integration they do not guarantee success in the application of an inter-provider critical path.

Sequence confirmation and characterization of the mouse Ssxa gene: Ssxa protein is cleaved and the N-terminal cleaved fragment translocates into the nucleus.

This is the first demonstration of a stop codon in the sequence of mouse Ssxa and characterization of the biological behavior of Ssxa protein. Cancer testis antigen (CTA) is known as a target of immunotherapy against cancer, and Ssxa is one of the CTAs. Although a CTA would be useful to establish a mouse cancer vaccine model using endogenous antigen, the stop codon was not identified in the sequence of Ssxa cDNA that was previously reported. In this study, the gene sequence of Ssxa was different from the previous report in which several mouse CTAs were analyzed. Initially, we identified the correct cDNA sequence of mouse Ssxa by 3'-rapid amplification of cDNA ends and found a new exon containing the stop codon (Exon X). Ssxa mRNA expression was determined by reverse transcription-PCR (RT-PCR) in four mouse cancer cell lines and the testis but not in other normal organs. We found that the molecular weight of recombinant Ssxa protein is 12 kDa, and we generated an anti-Ssxa antibody which recognizes the C-terminus of Ssxa. Two vectors expressing fusion proteins (pSsxa-GFP and pGFP-Ssxa) were generated and fluorescence in the nucleus was observed only in the pGFP-Ssxa transfected cells. Therefore, we conclude that the N-terminal cleaved fragment of Ssxa, which has a KRAB domain (nuclear localization signal), translocates into the nucleus after cleavage of the C-terminus.

The CD40-CD154 interaction would correlate with proliferation and immune escape in pancreatic ductal adenocarcinoma.

BACKGROUND: CD40 and CD154 are associated with lymphocyte signaling pathways and they are also expressed in some malignant neoplasms, but the significance in pancreatic cancer is unknown. METHODS: Eighty pancreatic cancer specimens were stained immunohistochemically, and the results were correlated with the patients' clinicopathologic features. Subsequently, in vitro analysis of CD40-CD154 signaling was performed. RESULT: Immunohistochemical analysis of tumor cells showed that 29 patients (36.3%) were positive for CD40, and 17 patients (21.3%) had very high CD154 expression. The survival of patients who had very high CD154 expression was significantly better than that of others (P = 0.0198). Univariate and multivariate analysis revealed that very high CD154 expression in cancer cells was not an independent, favorable prognostic factor (risk ratio, 0.493; P = 0.0224). On in vitro proliferation assay, the growth of PK-45P and KP-4 cells was blocked by CD40 and CD154 blocking antibodies. Moreover, on in vitro cytokine assay, Th-2 cytokines from PK-45P and SUIT-2 were blocked by CD40 or CD154 blocking antibody. CONCLUSION: These results suggest that the CD40-CD154 interaction would correlate with cell proliferation and secretion of cytokines in PDAC cells, and CD154 overexpression could be a favorable prognostic factor in PDAC patients.

Identification of a royal jelly glycoprotein that carries unique complex-type N-glycans harboring the T-antigen (Galß1-3GalNAc) unit.

In this study, we identified a royal jelly glycoprotein (RJG) that carries a unique complex-type N-glycans harboring the T-antigen (Galß1-3GalNAc) unit. The amino acid sequence of the tryptic glycopeptide harboring the T-antigen unit was G-E-S-L-X-K (X might be glycosylated Asn), confirmed in the major royal jelly glycoprotein 1 (MRJP1), which is also expressed in the mushroom body of the honeybee brain.

Efficacy of the addition of prokinetics for proton pump inhibitor (PPI) resistant non-erosive reflux disease (NERD) patients: significance of frequency scale for the symptom of GERD (FSSG) on decision of treatment strategy.

BACKGROUND: Non-erosive reflux disease (NERD) is a more difficult to treat than reflux esophagitis (RE) due to the high prevalence of PPI resistance. Consequently, the treatment strategy for NERD is yet to be established. PATIENTS AND METHODS: Subjects were 467 GERD patients (NERD 349, RE 118, 47.4 +/- 16.7 years) with reflux symptoms such as heartburn. PPI was administered for 2 weeks, and total score (TS) of symptoms, seven items of reflux symptoms e.g. heartburn (reflux score: RS), and five items of dyspeptic symptoms e.g. heavy stomach (dyspeptic score: DS) were assessed using the frequency scale for the symptoms of GERD (FSSG), a GERD-specific questionnaire developed in Japan. Improvement <50% in TS was defined as non-responder. Patients' background, and pretreatment TS, RS, DS, and 12 items of FSSG were assessed. Furthermore, the effect of additional prokinetics (4 weeks) for 117 PPI non-response NERD patients was also examined. RESULTS: Younger age, constipation, higher TS, DS, F2 (bloated stomach), 3 (heavy stomach), 5 (sick feeling after meal), 8 (satiety during meal) in FSSG were factors to be PPI non-responders in NERD. Significant improvement in TS were observed (pretreatment: 17.4 +/- 7.7 vs. 2 weeks 14.6 +/- 6.0 vs. 6 weeks 7.7 +/- 5.2, p<0.0001) after the addition of prokinetics in PPI non-response NERD. CONCLUSION: Younger age, constipation, dysmotility were factors of PPI non-response in NERD. As high DS is correlated with PPI non-response, it is indicated that patients with strong dysmotility and functional dyspepsia complication might be PPI resistant. The efficacy of additional prokinetics for PPI non-response NERD was observed.

Wnt inhibitor Dickkopf-1 as a target for passive cancer immunotherapy.

Dickkopf-1 (DKK1) is an inhibitor of Wnt/beta-catenin signaling that is overexpressed in most lung and esophageal cancers. Here, we show its utility as a serum biomarker for a wide range of human cancers, and we offer evidence favoring the potential application of anti-DKK1 antibodies for cancer treatment. Using an original ELISA system, high levels of DKK1 protein were found in serologic samples from 906 patients with cancers of the pancreas, stomach, liver, bile duct, breast, and cervix, which also showed elevated expression levels of DKK1. Additionally, anti-DKK1 antibody inhibited the invasive activity and the growth of cancer cells in vitro and suppressed the growth of engrafted tumors in vivo. Tumor tissues treated with anti-DKK1 displayed significant fibrotic changes and a decrease in viable cancer cells without apparent toxicity in mice. Our findings suggest DKK1 as a serum biomarker for screening against a variety of cancers, and anti-DKK1 antibodies as potential theranostic tools for diagnosis and treatment of cancer.

Prognostic impact of regional lymph node micrometastasis in patients with node-negative biliary cancer.

OBJECTIVE: To immunohistochemically identify regional lymph node micrometastases in patients with regional node-negative biliary cancer who underwent curative resection, and to evaluate their clinical significance. SUMMARY BACKGROUND DATA: The clinical significance of immunohistochemically detected lymph node micrometastasis has recently been evaluated in various tumors. However, few reports have focused on this issue with regard to biliary cancer. METHODS: A total of 1421 regional lymph nodes from 151 patients with biliary cancer with negative regional nodes (as determined by conventional methods) were immunostained with antibody against cytokeratins 7 and 8 (CAM5.2). Prognostic impact was evaluated among patients with no metastasis, micrometastasis, and obvious metastasis detected by hematoxylin and eosin staining. Immunostained tumor foci were classified as small micrometastasis or large micrometastasis according to size (above or below 0.2 mm). RESULTS: CAM5.2-positive occult carcinoma cells in regional lymph nodes were detected in 33 (22%) of 151 patients and 49 (3%) of 1421 regional lymph nodes. Small micrometastases were detected in 23 patients, whereas large micrometastases were found in 10 patients. Survival for patients with micrometastasis was significantly worse than that for patients without (P = 0.0051), but was significantly better than that for patients with overt metastasis (P = 0.0092). No significant difference in postoperative survival was seen between patients with small and large micrometastases (P = 0.4221). CONCLUSIONS: Occult cancer cells were present in regional lymph nodes of 22% patients with regional node-negative biliary cancer, and were associated with significantly worse survival. Patients with micrometastases should be treated as carefully as node-positive patients.

Aberrant expressions of HOX genes in colorectal and hepatocellular carcinomas.

HOX genes are known as master regulator genes which give cells positional information in embryogenesis. In this study, we compared the expression patterns of 39 HOX genes among human colorectal carcinomas from the right large intestine (cecum, ascending and transverse colon), those from the left large intestine (discending and sigmoid colon, and rectum) and hepatocellular carcinoma. The expression levels of each HOX gene were quantified by analysis based on the real-time RT-PCR. The expression patterns of HOX genes in colorectal and hepatocellular carcinoma tissues differed from those in their normal or non-cancerous tissues. Between the tumor tissues in the right-side large intestine and those in the left-side, different HOX genes were expressed in association with cancer. Further, the expression levels of HOXD8 in liver-metastatic tissues of colorectal carcinomas were as low as in non-cancerous liver tissues, and were significantly lower than those in the primary tissues. These results suggest that dysregulated expressions of HOX genes play an important role in carcinogenesis and malignant progression of colorectal and hepatocellular carcinomas.

Differential LC-MS-based proteomics of surgical human cholangiocarcinoma tissues.

Cholangiocarcinoma is an intractable cancer for which there is no effective therapy other than surgical resection, and many patients are not candidates for this treatment. Even for patients who undergo surgical resection, the 5-year survival rate is low. One reason for this is that the disease is often detected in late stages. Thus, there is a clear need for better biomarkers to facilitate early diagnosis and prognostication. During the biomarker discovery phase of our study, we used LC-MS-based proteomics with spectral counting, a semiquantitative approach to differential expression profiling, in paired cancerous and normal bile duct tissue samples from two cases. In total, 38 proteins up-regulated in the cancer samples were identified. These were verified using a SILAC method for MS-based validation. The results led to the identification of well-characterized proteins and proteins of unknown function that are up-regulated in cholangiocarcinoma. We used immunoblot analysis to validate four candidate biomarkers, actinin-1, actinin-4, protein DJ-1 and cathepsin B, with the test case samples and four additional cholangiocarcinoma case samples. Each of the four candidate proteins was overexpressed in a subset of five of the six cases tested. By immunohistochemistry, we further confirmed that expression of these proteins was elevated in cancer cells as compared with normal bile duct cells. Thus, we successfully identified several proteins up-regulated in cholangiocarcinoma. These proteins are candidate biomarkers and may also help to provide new insights into our understanding of the disease.