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Supplementary Data from Decreased ID2 Promotes Metastatic Potentials of Hepatocellular Carcinoma by Altering Secretion of Vascular Endothelial Growth Factor

Purpose: We aimed to explore the molecular and biological functions of Inhibitor of DNA binding/differentiation 2 (ID2), which was found to be responsible for portal vein invasion of hepatocellular carcinoma (HCC).Experimental Design: We measured ID2 mRNA levels in 92 HCC patients by real-time reverse transcription-PCR and examined the relation to clinicopathologic features. To clarify the precise roles of ID2, we did in vitro analysis with expression vectors and small interfering RNAs. Effects of ID2 on cell invasive potential and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α were analyzed by Matrigel-coated invasion chamber, ELISA, and Western blot analysis, respectively.Results:ID2 mRNA level correlated inversely with portal vein invasion (P < 0.001), tumor-node-metastasis stage (P < 0.001), tumor size (P < 0.001), and early intrahepatic recurrence (P < 0.05). When limited to a cohort of hepatitis C virus–related HCCs, patients with low levels of ID2 had significantly shorter disease-free survival time than those with high levels of ID2. Invasive potential of cells transfected with ID2 expression vector was lower than that of empty vector–transfected cells. Cells overexpressing ID2 also showed decreased VEGF secretion and hypoxia-inducible factor-1α protein levels. The results of ID2-knockdown experiments were opposite to those of ID2 overexpression experiments.Conclusions: On the basis of our clinical and in vitro data, we suggest that ID2 plays a significant role in the metastatic process during progression of HCC. This action might be explained, at least in part, by altered cell mobility due to decreased secretion of VEGF.

Supplementary Figures 1-3, Table 1 from Integrin α2 Mediates Selective Metastasis to the Liver

We discuss a novel diagnostic method for predicting the early recurrence of liver cancer with high accuracy for personalized medicine. The difficulty with cancer treatment is that even if the types of cancer are the same, the cancers vary depending on the patient. Thus, remarkable attention has been paid to personalized medicine. Unfortunately, although the Tokyo Score, the Modified JIS, and the TNM classification have been proposed as liver scoring systems, none of these scoring systems have met the needs of clinical practice. In this paper, we convert continuous and discrete data to categorical data and keep the natively categorical data as is. Then, we propose a discrete Bayes decision rule that can deal with the categorical data. This may lead to its use with various types of laboratory data. Experimental results show that the proposed method produced a sensitivity of 0.86 and a specificity of 0.49 for the test samples. This suggests that our method may be superior to the well-known Tokyo Score, the Modified JIS, and the TNM classification in terms of sensitivity. Additional comparative study shows that if the numbers of test samples in two classes are the same, this method works well in terms of theF1measure compared to the existing scoring methods.

We previously reported the effectiveness of the product of tumor number and size (NxS factor) for the prognosis of hepatocellular carcinoma (HCC) in patients following hepatectomy. The present study aimed to propose a new score based on the NxS factor to predict HCC recurrence following hepatectomy. A total of 406 patients who underwent hepatectomy for HCC at Osaka University Graduate School of Medicine were retrospectively analyzed to develop the new score. Among clinicopathological factors, including the NxS factor, the marker subset that achieved the best performance for prediction of early recurrence was assessed, and a prognostic model for HCC recurrence after curative hepatectomy (REACH) was developed. As the validation set, 425 patients who underwent hepatectomy for HCC at Yamaguchi University Graduate School of Medicine and Shimonoseki Medical Center were analyzed, and the prognostic ability of the REACH score was compared with that of well-known staging systems. Following analysis, the REACH score was constructed using six covariates (NxS factor, microscopic hepatic vein invasion, differentiation, serum albumin, platelet count and indocyanine green retention rate at 15 min). In the validation set, the REACH score predicted early recurrence in 73 of 81 samples, with a sensitivity of 89% and a specificity of 58%. The area under the curve (AUC) of the receiver operating characteristic curve of the REACH score was 0.78 and 0.74, respectively, for 1- and 2-year recurrence after hepatectomy; each AUC was higher than that of any of the other staging systems. Survival analysis indicated the REACH score had the best predictive value in disease-free and overall survival. The present findings demonstrated that the REACH score may be used to classify patients with HCC into high- and low-risk of recurrence, and to predict subsequent survival following hepatic resection.

The aim of this study was to develop an accurate predictive system for prognosis of hepatocellular carcinoma (HCC) patients after hepatectomy. We pooled data of clinicopathological features of 234 HCC patients who underwent curative hepatectomy. On the basis of the pooled data, we established a simple predictive staging system (PS score) scored by the mathematical product of tumor number and size, and degree of liver function. We compared the prognostic abilities of the PS score (score 0–3) with those of six well-known clinical staging systems. Then, we found that there were significant differences (P

A therapeutic vaccine against minimal residual cancer cells is needed for the treatment of patients with colorectal cancer. Several gene therapy studies have revealed that the combination of a suicide gene and cytokine gene might induce effective antitumor immunity. In this study, we constructed an interleukin (IL)-18 and herpes simplex virus-thymidine kinase (HSV-TK) expression vector driven by the human telomerase reverse transcriptase (hTERT) promoter to study the efficacy of combination gene therapy with IL-18 and the HSV-TK suicide gene. Low immunogenic colon 26 cells were used for transfection and inoculation into syngeneic BALB/c mice. Large established tumors of colon 26 transfectants expressing IL-18 and HSV-TK driven by the hTERT promoter were completely eradicated after GCV administration in syngeneic BALB/c mice. Immunohistochemical analysis at the tumor rejection sites revealed enormous infiltrations of CD8+ T lymphocytes as well as CD4+ T lymphocytes and CD11b+ monocytes. Moreover, established distant tumors were completely eradicated by vaccination with the IL-18 and HSV-TK transfectants in combination with GCV. These data suggest that the IL-18 and suicide gene therapy can elicit antitumor specific immunity. In conclusion, gene therapy with IL-18 and HSV-TK plasmid vector driven by the hTERT promoter may be useful for cancer vaccination.

Downregulation of inhibitor of DNA binding 2 (ID2) is associated with poor prognosis in cases of hepatocellular carcinoma (HCC). Therefore, to search for effective antitumor drugs for the treatment of HCC exhibiting poor prognostic indicators, we used two HCC-derived cell lines (HuH-7 and HLE) to alter ID2 levels. Specifically, ID2 expression was knocked down in HuH-7 cells via transfection with ID2-specific small interfering RNAs and separately ID2 was overexpressed in HLE cells via an ID2 expression plasmid vector. To assess the effect of antitumor drugs, MTS assay was performed. Annexin V staining was used to evaluate apoptosis and real-time RT-PCR was used to measure mRNA levels. ID2 knockdown cells were more susceptible to histone deacethylase (HDAC) inhibitors including sodium butyrate (NaB), sodium 4-phenyl-butyrate, tricostatin A, suberoylanilide hydroxamic acid, MS-275, apicidin and HC-toxin. Conversely, cells that overexpressed ID2 were less susceptible than control cells to HDAC inhibitors. NaB-induced apoptosis was inversely correlated with ID2 expression. Expression of the anti-apoptotic mRNA BCL2 was induced by NaB in control cells, but this induction of BCL2 was inhibited by ID2 knockdown and strengthened by ID2 overexpression. Expression of another anti-apoptotic mRNA, BCL2L1, was decreased by NaB administration and then partially recovered. However, in ID2 knockdown cells, BCL2L1 levels did not recover from NaB-induced suppression. ID2 affected the susceptibility of two HCC-derived cell lines to an HDAC inhibitor by regulating the expression of anti-apoptotic genes. Therefore, HDAC inhibitors may be effective for the treatment of HCC for which the prognosis is poor based on ID2 downregulation and ID2 could serve as a marker that is predictive of the clinical response to HDAC inhibitors.

Measuring the learning effectiveness of Kampo Medicine (KM) education is challenging. The aim of this study was to develop a web-based test to measure the learning effectiveness of KM education among medical students (MSs). We used an open-source Moodle platform to test 30 multiple-choice questions classified into 8-type fields (eight basic concepts of KM) including “qi-blood-fluid” and “five-element” theories, on 117 fourth-year MSs. The mean (±standard deviation [SD]) score on the web-based test was30.2±11.9(/100). The correct answer rate ranged from 17% to 36%. A pattern-based portfolio enabled these rates to be individualized in terms of KM proficiency. MSs with scores higher (n=19) or lower (n=14) than mean ± 1SD were defined as high or low achievers, respectively. Cluster analysis using the correct answer rates for the 8-type field questions revealed clear divisions between high and low achievers. Interestingly, each high achiever had a different proficiency pattern. In contrast, three major clusters were evident among low achievers, all of whom responded with a low percentage of or no correct answers. In addition, a combination of three questions accurately classified high and low achievers. These findings suggest that our web-based test allows individual quantitative assessment of the learning effectiveness of KM education among MSs.

DNA microarray technology has revolutionized our understanding of the molecular basis of hepatocellular carcinoma (HCC), one of the most fatal human cancers with a high recurrence rate. Many researchers have used DNA microarray technology to reclassify HCC with respect to metastatic potential and to develop predictors for the outcome of HCC. However, developed predictors have reached the level only of small retrospective studies, and their current status is far from that required for clinical use. This is due to the lack of transparent data, the high cost and data instability associated with the high dimensionality of the technique, the infancy of bioinformatics, and the complicated nature of recurrent HCC. This comprehensive review summarizes: (i) class comparison studies to identify genes or pathways involved in HCC metastasis (ii) class discovery studies that have resulted in the identification of a new molecular subclass of HCC with respect to metastasis, and (iii) class prediction studies to develop multidimensional predictors for HCC outcome. We also discuss issues that need to be addressed so that the power of array-based predictors can be estimated prospectively in large independent cohorts of HCC patients.

Purpose: We aimed to explore the molecular and biological functions of Inhibitor of DNA binding/differentiation 2 (ID2), which was found to be responsible for portal vein invasion of hepatocellular carcinoma (HCC). Experimental Design: We measured ID2 mRNA levels in 92 HCC patients by real-time reverse transcription-PCR and examined the relation to clinicopathologic features. To clarify the precise roles of ID2, we did in vitro analysis with expression vectors and small interfering RNAs. Effects of ID2 on cell invasive potential and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α were analyzed by Matrigel-coated invasion chamber, ELISA, and Western blot analysis, respectively. Results: ID2 mRNA level correlated inversely with portal vein invasion (P < 0.001), tumor-node-metastasis stage (P < 0.001), tumor size (P < 0.001), and early intrahepatic recurrence (P < 0.05). When limited to a cohort of hepatitis C virus–related HCCs, patients with low levels of ID2 had significantly shorter disease-free survival time than those with high levels of ID2. Invasive potential of cells transfected with ID2 expression vector was lower than that of empty vector–transfected cells. Cells overexpressing ID2 also showed decreased VEGF secretion and hypoxia-inducible factor-1α protein levels. The results of ID2-knockdown experiments were opposite to those of ID2 overexpression experiments. Conclusions: On the basis of our clinical and in vitro data, we suggest that ID2 plays a significant role in the metastatic process during progression of HCC. This action might be explained, at least in part, by altered cell mobility due to decreased secretion of VEGF.

We measured intersubband (ISB) absorption in high-mesa waveguides consisting of AlN cladding layers and AlN/GaN multiple quantum wells (MQW). Inductively coupled plasma (ICP) dry etching for waveguide fabrication was optimized to reduce propagation loss by achieving vertical and smooth sidewalls. We changed the ICP/RF power and introduced intervals during the etching process in order to overcome the induction time of the etching rate and the instability of plasma observed from long etching experiments. The ISB absorption greater than 20 dB was clearly observed at around 1.5 µm. The insertion loss of an 800-µm-long waveguide was 5–10 dB including the coupling loss between a fiber and the end of the waveguide. With this low-loss waveguide, we observed a saturation of 10 dB in the ISB absorption with 100 pJ probe light introduced to the waveguide. We thus demonstrated the possibility of achieving all-optical switching devices with low loss using the ISB transition of AlN-based materials.

In a previous study, we showed that levels of cell-free DNA (cfDNA) were significantly higher in sera of patients with hepatocellular carcinoma (HCC) associated with hepatitis C virus (HCV) than in sera of non-HCC patients with HCV. To confirm this finding, we analysed serum cfDNA levels in a cohort of 96 patients with HCV-related HCC and in 100 HCV carriers without known HCC. Again we found that serum cfDNA levels were significantly higher in HCC patients than in HCV carriers (115.9+/-98.3 vs 34.4+/-40.4 ng ml(-1) (mean+/-s.d.), P0.0001). Of 87 eligible patients who underwent curative hepatectomy, those with a high cfDNA level had a significantly shorter overall survival (OS) time than those in whom the cfDNA level was not high. Cox proportional hazards model showed the cfDNA level to be an independent prognostic factor for OS and cancer recurrence in distant organs. Our results suggest that the serum cfDNA level reflects the metastatic potential of HCV-related HCC and that it can be a useful predictive biomarker for distant metastasis after curative surgery.

We report the first successful realization of an AlN-waveguide-based intersubband transition device prepared using both metalorganic vapor phase epitaxy (MOVPE) and molecular beam epitaxy (MBE) techniques to grow AlN-cladding GaN/AlN quantum wells. An intersubband absorption wavelength of as short as 1.3 µm is confirmed with a new measurement method using an ultrawide-spectrum light source. With this method, the intersubband absorption spectra of an AlN high-mesa waveguide can be directly observed for the first time. The intersubband absorption saturation measurements also show a good device characteristic with a saturation of 7 dB for a transverse magnetic (TM) polarized input pulse energy of 200 pJ.

To identify proteins associated with esophageal carcinogenesis, we performed protein profiling of 16 esophageal squamous cell carcinomas (ESCCs) and paired noncancerous tissues by 2-DE and MS/MS. In cancerous tissues, three spots showed significant up-regulation in the amount of protein, while eight spots were significantly down-regulated. The identities of the spots were determined by PMF with LC-MS/MS and were confirmed by immunoblotting. The up-regulated proteins were tropomyosin alpha 4 chain, transgelin, and pyruvate kinase. The down-regulated proteins were serum albumin precursor, isoforms of annexin A1, tropomyosin beta chain, 14-3-3 protein sigma, and isoforms of serotransferrin precursor. In all 16 cases, up-regulation of the tropomyosin alpha 4 chain was confirmed by immunoblotting. Localization of the tropomyosin alpha 4 chain in ESCC cells and adjacent fibroblasts was confirmed by immunohistochemistry.

This study aimed to investigate the relationship between prognosis after curative hepatectomy and serum methylation signature (SMS), defined by methylation levels of six specific genes (cyclin D2, Ras association (RalGDS/AF-6) domain family member 1, serine peptidase inhibitor Kunitz type 2, cystic fibrosis transmembrane conductance regulator, brain abundant membrane attached signal protein 1, and steroid-5-alpha-reductase alpha polypeptide 2).Serum samples were collected preoperatively from 125 patients with hepatocellular carcinoma associated with hepatitis C virus infection who underwent curative hepatectomy. We measured the methylation levels of the preceding six genes. We defined the methylation of three genes or more in the serum as SMS-positive in this study. We investigated the prognosis of SMS-positive patients.SMS-positive patients exhibited significantly shorter disease-free survival (DFS) and overall survival (OS) than SMS-negative patients (p=0.0002 and p0.0001, respectively). Multivariate analysis showed that SMS positivity was an independent risk factor for shorter DFS (hazard ratio (HR)=2.182; p0.001) and OS (HR=4.198; p0.001).SMS is useful as a prognostic predictor in patients with hepatocellular carcinoma after curative hepatectomy.

We demonstrated high-speed, stable and unidirectional microring III-V laser diodes with in-cavity feedback silicon waveguide, and the integration with III-V photodetector on a silicon-on-insulator platform to achieve on-chip data transmission with a data rate of 12.5 Gb/s.

Intersubband transition (ISBT) in GaN quantum wells (QWs) was investigated from the viewpoint of application to ultrafast all-optical switches. The effect of crystalline quality on the absorption saturation characteristics was examined and reduction of edge dislocations was found to be a crucial factor in decreasing operation energy. Then, the switching performance was investigated for devices with improved crystalline quality. Modulation of signal pulses with a pulse interval of less than 1 ps was confirmed. Another device displayed gate-switch operation with an extinction ratio of greater than 10 dB. Comparison of the absorption recovery process in both devices suggested that the process is strongly affected by the QW structure

To detect autoantibodies that could be diagnostic markers for hepatocellular carcinoma (HCC), we analyzed serum autoantibodies comprehensively that showed immunoreactivity to proteins in tumor tissue obtained from patients with HCC. Fifteen paired samples of HCC tissue and corresponding nontumorous liver tissue as well as five normal liver tissue samples were used in the study. A combination of proteomics and SEREX (serologic analysis of recombinant cDNA expression libraries) technique was used. Tissue proteins were separated by 2-DE, transferred onto PVDF membranes, and immunoblotted with autologous sera. By comparing each immunoblot pattern, we identified four immunoreactive spots with stronger staining intensity in tumorous tissues than in corresponding nontumorous tissues and in normal liver tissues. Matched proteins on 2-DE gels were identified by LC-MS/MS. These immunoreactive proteins were heat shock 70 kDa protein 1 (HSP70), glyceraldehyde 3-phosphate dehydrogenase, peroxiredoxin, and manganese superoxide dismutase (Mn-SOD). In HCC sera, occurrences of autoantibodies against these proteins were 7/15 (46.7%), 5/15 (33.3%), 5/15 (33.3%), and 6/15 (40.0%), respectively, whereas 2/20 (10.0%), 7/20 (35.0%), 0/20 (0.0%), and 2/20 (10.0%) were in control sera. Immunoblot analysis using commercially available purified proteins was performed to confirm the specificity of autoantibodies. By statistical analysis, autoantibodies against HSP70, peroxiredoxin, and Mn-SOD showed significantly high-frequency immunoreaction in HCC sera. The three antibodies were considered patient-specific antibodies in HCC and may be candidate diagnostic biomarkers for HCC.

It is difficult to understand precisely the physiological actions of herbs because they contain a complex array of constituent molecules. In the present study we used DNA microarray data for 12600 genes to examine the anti-proliferative activity of the herb Coptidis rhizoma and eight constituent molecules against eight human pancreatic cancer cell lines. We identified 27 genes showing strong correlation with the 50% inhibitory dose (ID50) of C. rhizoma after 72-h exposure. Hierarchical cluster analysis with correlation coefficients between expression levels of these 27 C. rhizoma-related genes and the ID50 of each constituent molecule classified these test molecules into two clusters, one consisting of C. rhizoma and berberine and the other consisting of the remaining seven molecules. Our results suggest that one molecule, berberine, can account for the majority of the anti-proliferative activity of C. rhizoma and that DNA microarray analyses can be used to improve our understanding of the actions of an intact herb.

GaN was grown on a high-temperature AlN (HT-AlN) buffer by rf-MBE under various nitridation conditions (temperature, N2 flow rate, nitridation time and rf-power), and the dislocation density and the surface morphology were investigated. The edge dislocation density was decreased when the sapphire substrate was nitridated at a moderate temperature (540 °C). This can be explained by the difference in the density of the nuclei formed at the initial stage of the buffer layer growth. In addition, the edge dislocation density was found to be reduced by inserting a low temperature GaN buffer on the HT-AlN buffer. (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A one-dimensional finite-difference time-domain (FDTD) simulator for ultrafast optical switches based on intersubband transition (ISBT) in GaN/AIN waveguide is described. Influences of the inhomogeneous broadening and the 2D mode profile have been taken into consideration. The ultrafast optical response (τ ∼ 185 fs) measured in a GaN/AIN waveguide was successfully reproduced by the simulator. At present, however, the saturation characteristics of the fabricated device are mainly limited by the excess TM loss caused by the dislocation in MBE-grown nitride layers. When the dislocation density is reduced and the structure is optimized, the switching pulse energy will be improved to about 10 pJ. Further reduction (∼ 1 pJ) will be possible when low-loss submicron waveguides with spot-size converters are developed.

Using high-density oligonucleotide array, we comprehensively analyzed expression levels of 12600 genes in 50 hepatocellular carcinoma (HCC) samples with positive hepatitis C virus (HCV) serology (well (G1), moderately (G2), and poorly (G3) differentiated tumors) and 11 non-tumorous livers (L1 and L0) with and without HCV infection. We searched for discriminatory genes of transition (L0 vs. L1, L1 vs. G1, G1 vs. G2, G2 vs. G3) with a supervised learning method, and then arranged the samples by self-organizing map (SOM) with the discriminatory gene sets. The SOM arranged the five clusters on a unique sigmoidal curve in the order L0, L1, G1, G2, and G3. The sample arrangement reproduced development-related features of HCC such as p53 abnormality. Strikingly, G2 tumors without venous invasion were located closer to the G1 cluster, and most G2 tumors with venous invasion were located closer to the G3 cluster (P=0.001 by Fisher’s exact test). Our present profiling data will serve as a framework to understand the relation between the development and dedifferentiation of HCC.

To clarify the molecular basis of acquired cisplatin (CDDP) resistance in esophageal squamous cell carcinoma (ESCC), we used cDNA microarray technology. A CDDP-resistant cell line (YES-2/CDDP), which shows a 7.5-fold increase in resistance to CDDP and a 3-fold decrease in CDDP accumulation compared with the parental YES-2 ESCC cell line, was generated from YES-2 by exposure to increased concentrations of CDDP. By cDNA microarray analysis, we identified 44 genes with significantly different expression levels between YES-2/CDDP and YES-2 cells. Interestingly, 15 of these 44 genes encoded ribosome-related proteins, almost all of which were underexpressed in YES-2/CDDP cells. Our present data suggest that many ribosome-related genes may be involved in the acquired resistance to CDDP in ESCC and that such information may allow us to better understand the mechanism of CDDP resistance.

Intersubband transitions in GaN multiple quantum wells (MQW) are expected to be applicable to 1-Tb/s all-optical switches. The near-infrared (1.33–2.15 µm) intersubband absorption spectra of GaN/AlN MQW samples have been theoretically investigated. The measured spectra have been well explained by a model in which zero-voltage drop across one quantum well period and 0.5- to 0.75-nm-thick interface graded layers were assumed. The effective electric field in the well was estimated to be greater than 5 MV/cm. For thick wells, absorption spectra are sensitive to the electric field, and hence to the strain, barrier thickness, and doping level. In thin wells, the effect of the electric field is negligible, but the quality of the interface affects the spectra. The calculation model has been applied to the design of coupled quantum wells (CQW), where faster optical response is expected. Higher-order intersubband absorption at wavelengths of less than 1 µm is also expected to be enhanced in CQWs.

To clarify the roles of estrogen receptors (ERs) in gastric cancers, we evaluated expression of ER-alpha and ER-beta mRNAs in 41 pairs of tumorous and non-tumorous tissues of gastric cancer patients and in six gastric cancer cell lines by reverse transcription-polymerase chain reaction method. ER-alpha and ER-beta mRNAs were detected in 21 (51%) and 30 (73%) of 41 tumors and in 15 (37%) and 36 (88%) of 41 corresponding normal tissues, respectively. There were no statistically significant associations between expression of ER-alpha and/or ER-beta mRNAs in tumors and clinicopathologic factors. Between the tumorous and normal tissues, expression of ER-alpha and ER-beta mRNAs were changed in 20 (49%) and unchanged in 21 (51%) of the 41 cases. The incidences of lymph node metastasis and liver metastasis were significantly higher in changed cases than in unchanged cases (P=0.031 and P=0.021, respectively). We confirmed that ER-alpha and ER-beta mRNA were expressed in 2 and 6 of the six gastric cancer cell lines, respectively. Together with this finding, our results indicate that ER-beta mRNAs are preferentially expressed in gastric cancers. Our data also suggest that altered expression of ER-alpha and ER-beta mRNAs in tumors compared with corresponding normal gastric tissues is related to increased metastatic potential in gastric cancers. Further studies are needed to clarify the role of ERs in gastric cancers.

Constipation is characterized by a variety of bowel symptoms such as difficulty passing stool, hard stool, and a feeling of incomplete evacuation. The multifactorial causes of constipation limit the clinical efficacy of current conventional treatments that use a single drug that acts through only one pathway. To complement the shortcomings of the current Western medical model and provide a complete holistic approach, herbal medicines capable of targeting multiple organs and cellular sites may be used. In Japan, many herbs and herbal combinations have traditionally been used as foods and medicines. Currently, Japanese physicians use standardized herbal combinations that provide consistent and essential quality and quantity. This review highlights representative Japanese herbal medicines (JHMs), Rhei rhizoma-based JHMs including Daiokanzoto and Mashiningan, and Kenchuto-based JHMs including Keishikashakuyakuto and Daikenchuto, which coordinate the motility of the alimentary tract. This review provides a framework to better understand the clinical and pharmacological efficacies of JHMs on constipation according to the unique theory of Japanese traditional medicine, known as Kampo medicine.

We previously reported a phase I study of a cancer vaccine using five novel HLA-A*2402-restricted peptides, and demonstrated the safety and the promising potential of our five-peptide cocktail for advanced colorectal cancer. The objective of this analysis was to investigate predictive biomarkers for the prior selection of patients who are likely to have clinical benefit from such therapy.Seventeen patients with colorectal cancer who were treated with the five peptides underwent a complete blood count, serum chemistry tests and enzyme-linked ImmunoSpot assay before the treatment as predictive markers of high reactivity to the peptides.Interleukin-6 level was a significant predictor for overall survival of patients treated with the peptide cocktail (p=0.017). A high neutrophil/lymphocyte ratio was likely to have some association with the poor induction of peptide-specific immune reaction.Interleukin-6 level might be a good predictive biomarker for clinical benefit of patients treated with this peptide vaccine.

Liver cancer has a high likelihood of recurrence despite complete surgical resection and is thus known as an intractable cancer. If postoperative recurrence of cancer is correctly predicted for each patient as a form of personalized medicine, effective treatment can be carried out. The purpose of this paper is to evaluate classifiers for predicting recurrence of liver cancer by use of blood test data only in patients who underwent complete surgical resection of liver cancer. We conduct comparative study of 3 classifiers with use of binary patterns, which consist of clinical data and genomic data.

The contribution of the nm23-H1 gene to metastasis in malignant tumors, including gastric cancer, is controversial. In this study, we compared nm23-H1 levels in two cell subtypes with different morphologies (floating and adherent states), but that were derived from the same gastric cancer cell line, KATO-III. A real-time quantitative reverse transcription-polymerase chain reaction showed that the number of nm23-H1 mRNA molecules in floating cells was significantly higher than that in adherent cells (P

Ultrafast modulation of interband-resonant light by intersubband-resonant light in n-doped GaAs/AlGaAs and GaN/AlGaN quantum wells was investigated by femtosecond pump-probe technique. A planar-type AlGaAs/GaAs modulation device shows a modulation speed of ∼1 ps at room temperature. The observed modulation efficiency indicates that 99% modulation can be achieved with a control pulse energy of ∼1 pJ when a waveguide-type device structure is utilized. The feasibility of the all-optical modulation in GaN/AlGaN quantum wells is also investigated. The intersubband carrier relaxation time, which mainly determines the modulation speed, is measured and is found to be extremely fast (130–170 fs). The results indicate that the optical modulation at a bit rate of over 1 Tb/s will be possible by utilizing the intersubband transition in GaN/AlGaN quantum wells. The modulation efficiency in GaN/AlGaN quantum wells is also discussed in comparison with that in GaAs/AlGaAs quantum wells.

IL-18 is a novel cytokine that induces interferon (IFN)-gamma secretion and plays an important role in antitumor immunity. In the present study, we constructed plasmid vectors encoding the murine mature IL-18 cDNA linked with the Igkappa leader sequence and the pro-IL-18 cDNA to estimate the efficacy of the mature IL- 18 vector and to evaluate IL-18--producing tumor cells as a tumor vaccine. Colon 26 cells were transfected with the abovementioned vectors or with vector alone (mock). Reverse transcription-polymerase chain reaction analysis showed increased expression of murine IL-18 cDNA in both mature IL-18 and pro-IL-18 transfectants in comparison to that in mock transfected cells. The ability of the culture supernatants of mature IL-18 transfectants to induce IFN-gamma secretion was extremely high (40-140 pg/10(6) cells) in comparison to that of pro-IL-18 transfectants (4-18 pg/10(6) cells). When injected into syngeneic BALB/c mice, the growth of mature IL-18 transfectants, but not pro-IL-18 transfectants, was significantly less than that in mock transfected cells ( P< .01, by ANOVA and analysis of covariance). In addition, injection of colon 26 or Meth-A cells into mice immunized with a mature IL-18 transfectant revealed acquired immunity. Depletion of natural killer cells did not affect the growth of transfectants. However, the growth inhibitory effects were partially abrogated following treatment with anti-CD4+ and anti-CD8+ antibodies. These data suggest that the rejection of mature IL-18/colon 26 cells was mediated through T-cell activation. Gene therapy using mature IL-18 transfectants containing a plasmid vector and the Igkappa leader sequence may be a useful tumor vaccine.

Liver cancer has a high likelihood of recurrence despite complete surgical resection and is thus known as an intractable cancer. If postoperative recurrence of cancer is correctly predicted for each patient as a form of personalized medicine, effective treatment can be carried out. The purpose of this paper is to investigate prediction of recurrence of liver cancer by use of blood test data only in patients who underwent complete surgical resection of liver cancer. For this purpose, we propose a classifier based on Boolean algebra using a binary pattern consisting of a combination of clinical and genomic data by which we can predict recurrence of liver cancer. We perform a predictive experiment using data from patients with recurrence and non-recurrence and discuss the effectiveness of the proposed method from the experimental results.

Cytologic examination of peritoneal lavage fluid is a useful predictor of peritoneal recurrence in gastric cancer. However, this technique is not overly sensitive and requires special abilities in the cytologist. In this study, telomerase activity was used to detect free cancer cells in peritoneal lavage fluid from patients with gastric cancer. In the first part, 12 lavage-fluid samples obtained from 12 patients with gastric cancer were analysed using the conventional telomeric repeat amplification protocol (TRAP) assay. Three of five patients with early gastric cancer had positive telomerase activity. These false-positive results may have been due to lymphocyte contamination. Furthermore, polymerase chain reaction inhibitors were also detected in the lavage-fluid samples. Therefore, we developed a novel method for elimination of haematopoietic cell and Taq polymerase inhibitors to increase the accuracy of the TRAP assay using immunomagnetic beads, which bind to most normal and neoplastic human epithelial cells. Telomerase activity was found in 10 of 20 (50%) lavage-fluid samples from patients with serosal or subserosal invasion. Cytologic examination was positive in nine of 20 (45%) samples. Both the telomerase activity and cytology were negative in all 14 patients without serosal or subserosal invasion. These results suggest that the TRAP assay combined with immunomagnetic beads might be useful for detection of free cancer cells in the peritoneal space in gastric cancer without the aid of an experienced cytologist. © 2000 Cancer Research Campaign

Interleukin-1 receptor antagonist (IL-1ra), an endogeneous inhibitor of IL-1, plays an immunosuppressive role in vivo by blocking the proinflammatory effects of IL-1. In the present study, we examined whether IL-1ra expression in human gastric carcinoma correlates with tumor progression and/or metastatic potential. The reverse transcription-polymerase chain reaction was used to compare the expression of the secreted form of IL-1ra (sIL-1ra) and the intracellular form of IL-1ra (icIL-1ra) mRNA in tumor and corresponding benign tissue obtained from 38 patients with gastric carcinoma. The incidence of sIL-1ra mRNA expression was significantly higher in tumor (52%) than in corresponding benign tissue (18%) (P = 0.002). On the contrary, icIL-1ra mRNA was detected in all tumors and benign tissues. The expression of sIL-1ra mRNA by malignant tissue correlated positively with both lymph node metastasis (P = 0.008) and liver metastasis (P = 0.015). There was no association between tumor sIL-lra mRNA expression and other clinicopathologic factors. The degree of regional lymph node reaction, such as sinus histiocytosis, in tumors expressing sI-1ra mRNA was significantly weaker than that in tumors without sIL-1ra mRNA expression (5/20 vs. 12/18, P = 0.010). These results demonstrate that the altered expression of sIL-1ra by malignant tissue may be related to the progression of gastric carcinoma via modulating host immune response.