plus ATRA, and higher than those observed with arsenic trioxide plus ATRA. The secondary malignancy incidence after APL treatment was 2.7%, among the 75 patients that achieved CR, and 5.0% among the 40 patients that survived more than 5 years after the APL diagnosis. Conclusions: Adding cytarabine to anthracycline plus ATRA was not inferior to anthracycline plus ATRA alone, but it was not comparable to arsenic trioxide plus ATRA. The probability of secondary malignancy was low.

Premature fusion of the cranial suture and midface hypoplasia are common features of syndromic craniosynostosis caused by mutations in the FGFR2 gene. The only treatment for this condition involves a series of risky surgical procedures designed to correct defects in the craniofacial bones, which must be performed until brain growth has been completed. Several pharmacologic interventions directed at FGFR2 downstream signaling have been tested as potential treatments for premature coronal suture fusion in a mouse model of Apert syndrome. However, there are no published studies that have targeted for the pharmacologic treatment of midface hypoplasia. We used Fgfr2S252W/+ knock-in mice as a model of Apert syndrome and morphometric analyses to identify causal hypoplastic sites in the midface region. Three-dimensional geometric and linear analyses of Fgfr2S252W/+ mice at postnatal day 0 demonstrated distinct morphologic variance. The premature fusion of anterior facial bones, such as the maxilla, nasal, and frontal bones, rather than the cranium or cranial base, is the main contributing factor toward the anterior-posterior skull length shortening. The cranial base of the mouse model had a noticeable downward slant around the intersphenoid synchondrosis, which is related to distortion of the airway. Within a skull, the facial shape variance was highly correlated with the cranial base angle change along Fgfr2 S252W mutation-induced craniofacial anomalies. The inhibition of an FGFR2 downstream signaling enzyme, PIN1, via genetic knockdown or use of a PIN1 inhibitor, juglone, attenuated the aforementioned deformities in a mouse model of Apert syndrome. Overall, these results indicate that FGFR2 signaling is a key contributor toward abnormal anterior-posterior dimensional growth in the midface region. Our study suggests a novel therapeutic option for the prevention of craniofacial malformations induced by mutations in the FGFR2 gene.

Alveolar bone resorption caused by trauma or periodontal diseases has represented a challenge for both dental clinicians and researchers. In this study, we evaluate the osteogenic potential of human gingival fibroblasts (HGFs) through a direct transdifferentiation from HGFs to functional osteoblasts via epigenetic modification and osteogenic signaling with bone morphogenetic protein 2 (BMP2) in vitro and in vivo. HGF treatment with 5-aza-2'-deoxycytidine (5-aza-dC) induced demethylation in the hypermethylated CpG islands of the osteogenic lineage marker genes RUNX2 and ALP, and subsequent BMP2 treatment successfully drove the fibroblasts to the osteoblasts' lineage. Cell morphological changes viewed under microscopy and alkaline phosphatase (ALP) and alizarin red S (ARS) staining confirmed the osteoblastic change mediated by epigenetic modification as did real-time polymerase chain reaction (PCR), methylation-specific PCR (MSP), and chromatin immunoprecipitation (ChIP) assay, which demonstrated the altered methylation patterns in the RUNX2 and ALP promoter regions and their effect on gene expression. Furthermore, micro-computed tomography (CT) analysis of in vivo mouse cell transplantation experiments showed high-density signal in the epigenetically modified HGF group; in addition, a significant amount of bone formation was observed in the transplanted material using hematoxylin and eosin (H&E) staining as well. Collectively, our results indicate that epigenetic modification permits the direct programming of HGFs into functional osteoblasts, suggesting that this approach might open a novel therapeutic avenue in alveolar bone regeneration.

Previously, we found that osteogenic responses to zirconia co-doped with niobium oxide (Nb2O5) or tantalum oxide (Ta2O5) are comparable with responses to titanium, which is widely used as a dental implant material. The present study aimed to evaluate the in vitro osteogenic potential of hydroxyapatite (HA)-coated zirconia by an aerosol deposition method for improved osseointegration. Surface analysis by scanning electron microscopy and x-ray diffraction proved that a thin as-deposited HA film on zirconia showed a shallow, regular, crater-like surface. Deposition of dense and uniform HA films was measured by SEM, and the contact angle test demonstrated improved wettability of the HA-coated surface. Confocal laser scanning microscopy indicated that MC3T3-E1 pre-osteoblast attachment did not differ notably between the titanium and zirconia surfaces; however, cells on the HA-coated zirconia exhibited a lower proliferation than those on the uncoated zirconia late in the culture. Nevertheless, ALP, alizarin red S staining, and bone marker gene expression analysis indicated good osteogenic responses on HA-coated zirconia. Our results suggest that HA-coating by aerosol deposition improves the quality of surface modification and is favorable to osteogenesis.

Background: The novel hybrid retinoid, retinyl retinoate, is a synthetic material that was designed to reduce the side effects of retinoic acid and to increase the stability of retinol. The formulation of the retinyl retinoate, however, is required to enhance skin permeation, and thus to increase the anti-wrinkle effect.Aim: To identify the efficacy of retinyl retinoate microsphere using biodegradable polymer as an anti-aging agent of cosmetics in treating females over 30 years old with periorbital wrinkles.Methods: The retinyl retinoate microsphere was prepared using the biodegradable polymer; polylactic acid (PLA). We also conducted two clinical studies with a total of 44 Korean women for 12 weeks. In the first clinical study, 20 patients completed a 12-week trial of cream A [3% PLA-retinyl retinoate (2%) microsphere] applied twice daily to the face. In the second clinical study, 24 patients completed a 12-week trial of cream B (0.06% retinyl retinoate) applied twice daily to the face. Efficacy was based on a global photodamage score, photographs, and image analysis using replicas and Visiometers every 4 weeks.Results: The PLA-retinyl retinoate microsphere was more effective for the permeation of retinyl retinoate than retinyl retinoate in itself. The cream A, which contains 3% PLA-retinyl retinoate (2%) microsphere, showed a statistically significant improvement in facial wrinkles (P<0.05) in 20 volunteers after only 4 weeks of application in a clinical trial test. The visual wrinkle improvement and the maximum roughness improvement rate (R2) for cream A was 6.05%, 8.03% higher than that of cream B which contains 0.06% retinyl retinoate, after 4 weeks.Conclusion: Retinyl retinoate has a potent anti-wrinkle activity, and the PLA-retinyl retinoate microsphere could be a useful cosmeceutical product for anti-aging purposes.

ABSTRACT While the identification of cancer stem cells as therapeutic targets is now actively being pursued in a variety of human malignancies, leukemic stem cells (LSCs) in acute myeloid leukemia (AML) are a paradigm of such a strategy. However, significant improvements in outcome based on these insights have not been forthcoming, because the molecular basis to distinguish the properties of LSCs from those of normal hematopoietic stem cells (HSCs) remains largely unknown. Meanwhile, current advances in single-cell gene profiling can provide definitive evidence regarding the conversion of one cell type into another at a high level of resolution. To characterize biological subtypes of LSCs in AML, in this study, we systematically analyzed the cDNA profiles of a large number of single LSCs. We first carried out the comparative DNA microarray analysis with primary leukemic cells and reconstituted cells in xenograft, and extracted 22 intrinsic genes from 5,599 genes, of which expression was significantly up-regulated in reconstituted cells, as the markers of high grade, aggressive AML. Among them, Evi1 and MN1, both previously identified potential poor prognostic factors in AML, were included. Next, we isolated BM CD34 + 38- LSCs from refractory AML patients, randomly picked up single cells, and amplified a total of 96 single-cell cDNA samples from each patient, as well as CD34 + 38- cells derived from a healthy volunteer to obtain information from normal HSCs. By analyzing expression-patterning of the 22 identified genes in each cell, we have identified a set of genes, of which expression pattern was significantly correlated with that of Evi1. This correlation was not observed in their normal counterparts, indicating that it was specific for AML LSCs. Furthermore, in one case, who developed AML from myelodysplastic syndrome (MDS), the hierarchical cluster based on the molecular signatures was identified only in LSCs at AML stage, but not at MDS. These results suggest that single-cell gene-expression profiling would allows us to classify AML-LSCs and provide new insights into functional properties of LSCs. Currently, we are comprehensively evaluating the expression of surface molecules on AML LSCs, and would like to disclose the clinical practicality of this methodology in our presentation.

e18514 Background: We investigated the prognostic impact of bone marrow (BM) involvement in patients with diffuse large B-cell lymphoma (DLBCL) who received rituximab-based therapy. METHODS Thus, 567 consecutive patients with newly diagnosed DLBCL and treated with rituximab-CHOP (RCHOP) between August 2003 and December 2009 were included in the current study. All the patients underwent a BM study at the initial staging and the clinical characteristics and prognosis of these patients with or without BM involvement were retrospectively analyzed. RESULTS The total cohort included 567 patients. The median age was 61.0 years (range, 16-89) and 56.3% were male. Two hundred thirty patients (40.6%) had stage III and IV at diagnosis. One hundred seventy-seven patients (31.2 %) were categorized as high or high-intermediate risk according to their IPI. The overall incidence of BM involvement was 8.5%. A high number of abnormalities in the CBC profile was associated with a higher incidence of BM involvement (P<0.001). With a median follow-up duration of 33.2 months (range, 0.1-80.7) for the patients alive at the last follow-up, the 5-year overall survival (OS) and event-free survival (EFS) rate in the patients without BM involvement (67.5% and 76.3%) was statistically higher than that in the patients with BM involvement (40.1% and 44.3%). In a multivariate analysis, BM involvement was significantly associated with OS and EFS. CONCLUSIONS BM involvement at diagnosis affected the survival of the patients with DLBCL who received RCHOP. Although RCHOP can significantly improve the therapeutic effect of the DLBCL, BM involvement is still a negative prognostic factor of DLBCL patients in the era of rituximab.

280 Background: This phase II study evaluated efficacy of fixed dose rate (FDR) infusion of gemcitabine (10mg/m2/min) and UFT combination in chemo-naïve patients with advanced bile duct cancer. Methods: This was an open-label, single-arm, multicenter, phase II study with a Simon two-stage minimax design. Patients received the FDR gemcitabine 1,000mg/m2 for 3 consecutive weeks and UFT 400 mg/m2 on days 1-21. The cycle was repeated every 28 days. The primary end point was Response Evaluation Criteria in Solid Tumors (RECIST) -defined objective response rate. Secondary end points included clinical benefit response (CBR), safety, progression-free survival (PFS), and overall survival (OS). Clinical characteristics including four single nucleotide polymorphisms in DNA repair genes (RecQ1, RAD54L, XRCC1, ATM) were evaluated whether these influence the overall survival. Results: Between December 2006 and February 2008, fifty-one patients were enrolled, with a median age of 58 years. The majority of patients (76%) had intra-hepatic disease. Fourteen patients (27%) had a RECIST investigator-assessed, partial response (PR); disease control rate (PR + stable disease) was 55%. CBR was 14% among 37 evaluable patients. Hematologic toxicity was main grade 3 or 4 treatment-related adverse events. Median PFS was 4.0 months (95% CI, 2.9 to 5.1 months). Median OS was 7.0 months (95% CI, 3.5 to 10.5 months). Intrahepatic disease, poor performance, and, XRCC1 R194W C/C type were predictive markers of poor overall survival. Conclusions: FDR gemcitabine and UFT demonstrated apparent activity in patients with advanced bile duct cancer. However, this activity did not translate to prolong survival. The location of disease, performance status, and, polymorphic variants of DNA repair genes may affect clinical outcome of patients with advanced bile duct cancer. No significant financial relationships to disclose.

7603 Background: To compare the efficacy of gefitinib versus pemetrexed as second-line therapy in never-smokers with advanced pulmonary adenocarcinoma previously treated with platinum-based chemotherapy. Methods: Eligible patients had a performance status 0 to 2, previous treatment with one prior platinum-based regimen, pulmonary adenocarcinoma, and never-smoking state. Epidermal growth factor receptor (EGFR) mutation status was not considered for the enrollment. Randomized patients received either gefitinib 250 mg per oral every day or pemetrexed 500 mg/m2 iv day 1 with vitamin B12 and folic acid supplement every 21 days. The primary end point was progress-free survival (PFS). Results: One hundred thirty-five patients were randomly assigned with being stratified with performance status (0-1 vs. 2) and gender. Overall response rates were 30.1% and 14.9% (P < 0.001) for gefitinib and pemetrexed, respectively. The primary endpoint of PFS was met with 9.4 months for gefitinib versus 2.9 months for pemetrexed, which was significantly different (P = 0.010). The median overall survival has not been reached yet in both groups. The 1-year survival rate for gefitinib and pemetrexed arm was 73.6% and 70.5% (P = 0.89), respectively. The further analyses of EGFR mutation status, post-study treatment and adverse events will be presented on the ground. Conclusions: Gefitinib showed superior efficacy to pemetrexed as second-line therapy in clinically selected NSCLC patients in Korea. Considering sequence of salvage therapy, gefitinib would be preferable to pemetrexed in enriched NSCLC patients.

6524 Background: Although busulfan-cyclophosphamide (BuCy) has been commonly used as a myeloablative conditioning (MC) regimen for allogeneic hematopoietic cell transplantation (HCT), a new MC regimen using busulfan-fludarabine (BuFlu) showed less regimen-related toxicities and lower non-relapse mortality (NRM) compared to BuCy in several non-randomized studies. Thus, we performed a prospective, multicenter, open-label, phase III trial to compare BuCy and BuFlu as a MC regimen for allogeneic HCT in leukemia and myelodysplastic syndrome. METHODS A total of 130 were enrolled between Nov 2005 and Sep 2009, but 4 patients were removed from the study and the remaining 126 patients were analyzed. Median age was 41 years (range, 17 to 59). Underlying disease was as followings: AML 70, ALL 47, CML 2, MDS 6, MDS/MPN 1. After random assignments, 64 patients received busulfan (3.2mg/kg/d x 4d) plus cyclophosphamide (60mg/kg/d x 2d) [BuCy arm] and 62 received busulfan plus fludarabine (30mg/m2/d x 5d) [BuFlu arm]. RESULTS Baseline patient and donor characteristics were well balanced between two arms. Five patients experienced engraftment failure (primary 1, secondary 4), which occurred only in BuFlu. Incidence and severity of acute GVHD, chronic GVHD, hepatic SOS, CMV disease, interstitial pneumonitis, and hemorrhagic cystitis were not significantly different between two arms. Adverse events (AE) within 100 days after HCT were graded using NCI CTCAE v3.0. Both upper and lower gastrointestinal (GI) AEs were more frequent in BuCy compared to BuFlu (P=0.018 for upper GI, P=0.050 for lower GI). NRM was similar between BuCy and BuFlu (13.4% vs. 22.3% at 1 year, P=0.291), whereas relapse-free survival in BuCy was higher than that in BuFlu (67.7% vs. 50.7% at 4 year, P=0.039). Thus, overall and event-free survivals in BuCy were significantly higher than those in BuFlu (overall, 51.1% vs. 36.1%, P=0.008; event-free, 46.2% vs. 33.3%, P=0.023). CONCLUSIONS BuFlu caused less GI toxicities than BuCy, but NRM was similar. BuFlu showed lower survivals than BuCy mainly due to higher incidence of relapse. BuFlu cannot replace BuCy in patients who are eligible to myeloablative conditioning therapy before allogeneic HCT.

BACKGROUND:All-trans-retinoic acid (RA) and all-trans-retinol (ROL) are not widely used as anti-wrinkle agents due to their irritancy and photo-stability, respectively. Therefore, the safety and photo-stability in the development of RA or ROL derivatives have been an important issue.AIM:To identify the efficacy of retinyl retinoate as an anti-aging agent of cosmetics in treating females over 30 years old with periorbital wrinkles.METHODS:The clinical study was a prospective, double-blind, randomized, and controlled study with a total of 11 Korean women. At every 4 weeks, the effectiveness was assessed with a global photodamage score, photographs, and image analysis using replicas and visiometers. The dermal distance and intensity was also evaluated using Dermascan C.RESULTS:A statistically significant improvement in facial wrinkles (P<0.05) in eleven volunteers was observed in a clinical trial. The successive application of 0.06% retinyl retinoate cream for 3 months showed decreased depth and area of wrinkles in comparison with 0.075% retinol cream. The visual wrinkle improvement and the maximum roughness improvement rate (R2) for retinyl retinoate cream were 22% higher than that of retinol cream after 12 weeks. A statistically significant increase was observed after 8 and 4 weeks for dermal distance and dermal intensity, respectively (P<0.05).CONCLUSIONS:Retinyl retinoate had characteristic features of new anti-aging agents, and effectively improved facial wrinkle conditions.

The proteasome plays a pivotal role in controlling cell proliferation, apoptosis, and differentiation in a variety of tumor cells. Bortezomib is a boronic acid dipeptide derivative, which is a selective and potent inhibitor of the proteasome and has prominent effects in vitro and in vivo against several solid tumors. We examined the anti-proliferative and apoptotic effects of bortezomib in three gastric cancer cell lines (SNU638, MUGC-3 and MKN-28), along with its antitumor combination effects with other chemotherapeutic agents. Tumor cell growth inhibition and apoptosis was measured by MTT assay and FACS analysis, respectively. Apoptosis- and cell cycle-associated protein expression levels were measured by Western blot assay. Bortezomib induced the suppression of tumor cell growth and apoptosis in a dose-dependent manner with an inhibitory dose (ID)50 of approximately 0.5 microg/ml in all gastric cancer cell lines tested. Further combination treatment with cisplatin and docetaxel, in particular with docetaxel displayed dramatically increased tumor cell growth suppression in all three gastric cancer cell lines, as compared to single drug treatment alone. This was concomitant with the induction patterns of apoptotic cells. Bortezomib treatment increased the Bax protein expression. Moreover, combination treatment of bortezomib plus docetaxel resulted in a dramatic increase in the Bax expression. In contrast, Bcl-2 expression was decreased by combination treatment with bortezomib plus docetaxel in SNU638 cells. Finally, bortezomib, docetaxel and to a greater degree bortezomib plus docetaxel increased the expression levels of p27 proteins even without influencing p53 expression levels. Bortezomib has profound effects on tumor cell growth inhibition and induction of apoptosis in human gastric cancer cells, suggesting that bortezomib may be an effective therapeutic drug for patients with gastric cancer. Further combination studies with other chemotherapeutic drugs, in particular docetaxel showing more tumor cell growth inhibition and apoptosis suggest that combining bortezomib with docetaxel might be more effective for displaying tumor cell growth inhibitory effects in gastric cancer cells through regulation of Bcl-2, Bax and p27 proteins in vitro.

e19566 Background: A majority of patients with cancer experience cancer pain of higher than moderate severity during the progression of cancer. Of these, 2/3 of patients with cancer have been reported to be inflicted with breakthrough pain. Methods: The primary objective is to evaluate the clinical usefulness of hydromorphone OROS in reduction of breakthrough pain medication frequency in Korean patients. Subjects include patients who are given strong oral narcotic analgesics for the treatment of cancer pain. This study enrolled only patients who took immediate-release analgesics more than twice a day on average for the treatment of breakthrough pain during a 3-day period prior to Visit 2. When patients selected, patients were switched to test drug using a 2.5: 1 conversion ratio of controlled release oxycodone equivalent to hydromorphone hydrochloride, with no washout or overlap of previous opioid therapy and study medication. Results: Total 117 patients received the investigational drug and only 98 patients completed the last fourth assessment. The frequency of the use of immediate- release analgesics for the management of breakthrough pain was decreased from 2.93 times to 2.00 times (p<0.000). The incidence of breakthrough pain was also decreased from 3.67 times to 2.44 times (p<0.0001). K-BPI score was also decreased from 3.6 to 3.2. In 61.2% of patients had a satisfaction with the pain treatment. In regard to the preference to test drug in 83.7% of patients who completed the treatment, 82.95% of patients preferred test drug because ‘the cancer pain could be controlled with a one-time treatment'. 25.5% of patients showed improvement of CGI-I. In EORTC QLQ-C30 analysis, only pain and diarrhea among the symptom scales improved. In 91.2% of patients who used analgesics, the side effects occurred in such an order as constipation, drowsiness and dizziness. But there were no cases of uncontrolled specific side effects. Conclusions: A 24-hr sustained-release type of hydromorphone OROS tablet significantly diminished the use of immediate-release analgesics and the incidence of breakthrough pain. No significant financial relationships to disclose.

Purpose: This study was conducted to confirm the efficacy and toxicity of docetaxel and cisplatin combination chemotherapy (DP) in patients with advanced gastric cancer. Materials and Methods: Patients with measurable gastric adenocarcinoma received intravenous docetaxel 75 mg/m and cisplatin 75 mg/m with premedication on day 1, which was repeated every 3 weeks. All patients received DP as a second-line treatment after failing to 5-FU based chemotherapy. Results: 34 patients were enrolled in this study between January 1998 and August 2003. A total of 112 cycles (median 3 cycles) were administered. Responses were evaluable in 30 patients. The objective response rate was 16.7% (95% CI: 3.5~30.3), with a stable disease in 56.7% (95% CI: 40.0~74.4) and a progressive disease in 26.7% (95% CI: 10.9~42.5) of patients, with a median follow up duration of 20 months for all the patients, The median duration of response, time to progression and overall survival were 2.1 months (95% CI: 0.4~3.9), 4.2 months (95% CI: 2.3~6.1) and 6.8 months (95% CI: 1.3~12.3), respectively, with a 1-year survival rate of 32% . The toxicity was evaluated in 30 patients, with neutropenia being most common. Renal impairment was seen in two patients with grade 3 creatinine elevation and liver enzyme elevation in four with grades 3 and 4. Conclusion: Although DP was an active combination regimen, with a tumor control rate of about 73% and with moderate tolerance, adjustment of the administration schedule, with further evaluation of other combination chemotherapies of docetaxel with new agents, other than cisplatin, seem warranted. (Cancer Research and Treatment 2004;36:367-371) 󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏 Key W ords: Stomach neoplasm, Cisplatin, Docetaxel

BACKGROUND:Photoaged skin can be treated with retinoids, which are natural and synthetic vitamin A derivatives. However, these are photounstable and can cause skin irritation, which is a major limitation in their use in general cosmetics. Retinyl retinoate, which is an ester of all-trans retinoic acid (RA) and all-trans retinol, has reduced toxicity due to blocking of the carboxyl end group of RA and higher skin regeneration activity than retinol.OBJECTIVES:To assess the efficacy of a photostable retinyl retinoate in treating women over 30 years old with periorbital wrinkles.METHODS:We conducted two clinical studies with a total of 46 Korean women with periorbital wrinkles, who were not pregnant, nursing or undergoing any concurrent therapy. In the first clinical study, the efficacy of retinyl retinoate was compared with placebo. Twenty-four patients completed a 12-week trial of 0.06% retinyl retinoate applied twice daily to one side of the face and a placebo applied to the other side. In the second clinical study, the efficacy of retinyl retinoate was compared with retinol. Twenty-two patients completed an 8-week trial of 0.06% retinyl retinoate applied twice daily to one side of the face and 0.075% retinol applied to the other side. Efficacy was based on a global photodamage score, photographs, and image analysis using replicas and visiometer analysis (Skin-Visiometer SV 600; Courage & Khazaka, Cologne, Germany) every 4 weeks. The standard wrinkle and roughness parameters used in assessing skin by visiometer were calculated and statistically analysed.RESULTS:The retinyl retinoate-treated wrinkles improved compared with wrinkles treated with placebo or retinol, as assessed by both the investigators and the subjects. Also, skin replica analysis indicated significant improvements in retinyl retinoate-treated skin in both studies, especially in average roughness.CONCLUSIONS:Retinyl retinoate applied twice daily was significantly more effective than a placebo or retinol in treating periorbital wrinkles. Importantly, no severe side-effects were observed.

Spiro-fused heterocycles were synthesized in good to high yields by a pseudo four-component reaction of an aldehyde, urea and a cyclic β-diester or a β-diamide such as Meldrum’s acid or barbituric acid derivatives using microwave irradiation under solvent-free conditions.