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Carbonate apatite (CO3 Ap), an inorganic component of human bone, has been clinically applied as an artificial bone substitute. In this study, the effects of the CO3 content in CO3 Ap on the replacement by new bone were studied by fabricating CO3 Ap granules containing 0.9-8.3 wt% of CO3 . The dissolution rate of CO3 Ap in a weak acidic solution, mimicking the Howship's lacunae, was rapid for the CO3 Ap granules containing a larger amount of CO3 . Histological analyses demonstrated the rapid resorption in CO3 Ap and replacement by natural bone tissue when the CO3 content was increased. Therefore, the CO3 content in CO3 Ap is a key factor that influences the replacement of the bone tissue. This article is protected by copyright. All rights reserved.

Background The presence of uterine endometrial polyps is associated with not only abnormal uterine bleeding but also infertility, so the use of hysteroscopic polypectomy has been increasing. This operation is considered to increase cost-effectiveness when performed prior to infertility treatments. However, there are typical problems to consider, including the possibility of spontaneous regression of the polyp and the duration of complete endometrial wound healing after surgery. Meaningless interventions must be avoided, when possible. Therefore, data acquisition and analysis of various findings obtained from surgery have become important for improving treatment procedures and patient selection. To estimate the spontaneous regression rates and contributions of multiple factors to uterine endometrial polyps during the waiting period (approximately 2–3 months) before hysteroscopic polypectomy, we performed a multivariate analysis of data from the records in our hospital. Methods The medical records of 450 cases from September 2014 to April 2021 in our hospital were retrospectively reviewed under the approval of our Institutional Review Board. We included all cases of hysteroscopic polypectomy with postoperative pathological diagnosis. We defined cases as having a “spontaneously regressed polyp” when the target polyp was not detected by postoperative pathological examination. We extracted data on the following ten factors: “Advanced age” (≥ 42 years), “Small polyp” (2), “Nulliparity,” “Single polyp,” “Infertility,” “Hypermenorrhea,” “Abnormal bleeding,” “No symptom,” and “Hormonal drug use.” We also classified cases into five groups according to the size of the polyp (≤ 4.9 mm, 5.0–9.9 mm, 10.0–14.9 mm, 15.0–19.9 mm, and ≥ 20.0 mm) and determined the frequency of spontaneously regressed polyp in each group. Results After exclusion of cases with insufficient data or other diseases, such as submucosal leiomyoma, 424 cases were analyzed. Among them, 28 spontaneously regressed polyps were identified, and the highest frequency of spontaneously regressed polyp was detected among the cases with polyps measuring 5.0–9.9 mm (16.4%). On multivariate analysis of the ten factors, “Small polyp” and “Hormonal drug use” were found to significantly impact the frequency of spontaneously regressed polyp. Conclusions On the basis of the factors identified in this analysis, the indications for observation or medical therapy adapted to small polyps might be expanded.

Background The application of laparoscopic surgeries has been increasing, and various uterine diseases in addition to leiomyoma/adenomyoma have become indications for total laparoscopic hysterectomy (TLH). Therefore, data acquisition and analysis of TLH procedures, including TLH for rare uterine diseases, have become important for improving surgical procedures and patient selection. To determine the prevalence of and risk factors for the occurrence of intraoperative and postoperative complications of TLH, we performed a multivariate analysis of the records in our hospital. Methods We retrospectively reviewed the medical records of 323 patients who underwent TLH for the treatment of leiomyoma/adenomyoma (278 cases), low-grade (pre)malignant uterine tumours (40 cases) and other rare uterine diseases (5 cases) from January 1, 2015, to December 31, 2019. Of the rare uterine diseases, one case of caesarean scar pregnancy for which TLH was performed is introduced as a case report. To assess the effects of 11 representative factors, including patient characteristics, uterus and leiomyoma sizes, indications for TLH and others, we performed a multivariate logistic regression analysis. Results Among the 323 cases, 20 intraoperative complications and 15 postoperative complications were reported. In the multivariate analysis, “ovarian tumour” and “heavy uterus” were positively associated and “nulliparity” was negatively associated with intraoperative complications. There were no significant risk factors for postoperative complications. The only risk factor for operative complications directly related to the resected uterus was “heavy uterus”. Therefore, we could perform TLH relatively safely for patients with other indications besides leiomyoma/adenomyoma. Conclusions Considering the factors detected in this analysis, the indications for TLH may be expanded. Owing to the increase in TLH for indications other than leiomyoma/adenomyoma, a more accurate determination of the treatment approach can be achieved.

Background Acute puerperal uterine inversion is rare but may cause massive postpartum blood loss due to uterine atony. Therefore, these patients must be diagnosed, and uterine replacement must be performed as soon as possible. However, in some cases, active bleeding due to uterine atony becomes uncontrollable, even though the uterine inversion itself is treated. In these cases, additional treatments, including surgical procedures, are needed. Case presentation A 41-year-old Japanese woman, gravida 1, para 0, was hospitalized for labor induction at 40 weeks and 3 days of gestational age. She had a vacuum-assisted delivery after 3 days of oxytocin administration, but acute uterine inversion occurred. Although replacement of the inverted uterus was successful by manual repositioning and Bakri balloon tamponade insertion, massive postpartum hemorrhage caused by uterine atony became uncontrollable. In this situation, since disseminated intravascular coagulation had developed, we used uterine artery embolization to stop the bleeding. After detecting the pseudo-aneurysmal sac and tortuous vessels of the right uterine artery, transcatheter right-sided uterine artery embolization was performed. Thirteen days after uterine artery embolization, she was discharged with no complications. Conclusions In cases of disseminated intravascular coagulation caused by massive postpartum bleeding, uterine artery embolization may often be selected. In our case, since we performed angiography to detect the main bleeding site, the hemorrhage could be stopped with unilateral uterine artery embolization alone, without hysterectomy.

Background Etelcalcetide is a second-generation calcimimetic for the management of secondary hyperparathyroidism (SHPT) in patients on dialysis. We performed a post-marketing surveillance (PMS) to obtain information on the safety and efficacy of etelcalcetide in clinical practice in Japan. Methods This PMS enrolled SHPT patients who started initial treatment with etelcalcetide between April 1, 2017 and February 28, 2018 in Japan. Safety [adverse drug reactions (ADRs)] and efficacy [serum intact parathyroid hormone (iPTH), corrected calcium (cCa), phosphorous (P), and alkaline phosphatase (ALP)] were recorded for up to 52 weeks or until treatment discontinuation. Treatment decisions were at the physician’s discretion. Results Of 1226 patients enrolled across 282 centers, safety and efficacy data were available for 1195 and 1192, respectively, while 933 continued treatment to Week 52. The starting dose was 5 mg in 82.0% of patients. There were 218 ADRs in 169 patients (14.1%). Metabolism and nutrition disorders (8.8%), adverse laboratory test results (1.8%), and gastrointestinal disorders (1.6%) were the most frequent classes of ADRs. Hypocalcemia-related ADRs occurred in 104 patients (8.7%). The percentage of patients with iPTH levels within the target range (60–240 pg/mL) steadily increased from 19.5% at Week 0 to 64.1% at Week 52 or last dose. cCa, P, and ALP levels remained well controlled. Conclusion This was the first real-world, large-scale, long-term observational PMS of etelcalcetide in Japan. We did not observe any new safety concerns. Etelcalcetide was associated with clinically relevant improvements in serum iPTH and maintenance of serum cCa, P, and ALP levels.

We report 4 cases of liver metastasis from renal cell carcinoma(RCC). Case 1: 72 years old, female. Pancreatic metastasis was resected 7 years after resection of left RCC, and hepatic posterior sectionectomy was performed for multiple liver metastases 2 years later. After that, multi-organ metastasis appeared and she died of the primary disease. Case 2: 72 years old, male. Liver metastasis and right RCC appeared 16 years after resection of left RCC, and hepatic posterior sectionectomy and partial resection of right kidney were performed. Nine months later, liver metastasis recurred and hepatic partial resection was performed. Case 3: 55 years old, male. After surgery for right RCC with tumor thrombus in the right atrium, multiple lung and liver metastases appeared, and hepatic central bisectionectomy was performed after chemotherapy. Case 4: 60 years old, male. Multiple pancreatic and lung metastases appeared 10 years after resection of left RCC, and most of them shrank or disappeared with chemotherapy. But increasing metastases appeared in the tail of pancreas and the right lobe of liver 16 months later, and hepatic subsegmentectomy and distal pancreatectomy were performed. Multidisciplinary treatment such as aggressive chemotherapy and excision is expected to improve the prognosis for liver metastasis from RCC.

We investigated the significance of transitions in the neutrophil-to-lymphocyte ratio(NLR)before and after TACE for HCC could be a predictor of prognosis. The subjects were 108 patients with the first TACE performed from January 2010 to December 2019. NLR was calculated before and 1 month after TACE, and the relationship with therapeutic effect and prognosis was examined. When the transition of NLR before and after TACE was classified into 3 groups with a cut-off value of 5.0, group A(less than 5.0 after TACE): 52 cases(48.1%), group B(5.0 or more after TACE): 33 cases(30.6%)and C group(5.0 or more before and after TACE): 23 cases(21.3%). Median survival time were 25.0 months in group A, 18.5 months in group B, and 12.7 months in group C(p=0.0005). In multivariate analysis, treatment effect, NLR transition, AFP value, and serum albumin level were prognostic factors for HCC after TACE. Changes in NLR before and after TACE may help predict more detailed prognosis.

White mineral trioxide aggregate (WMTA) is a root canal treatment material, which is known to exhibit a dark brown color when in contact with sodium hypochlorite solution (NaOCl). This study aimed to investigate the effects of NaOCl on the surface properties of WMTA discs and WMTA-induced osteoblastic differentiation of periodontal ligament stem cells (PDLSCs). Mixed WMTA (ProRoot MTA) was filled into the molds to form WMTA discs. These discs were immersed in distilled water (D-WMTA) or 5% NaOCl (Na-WMTA). Their surface structures and Ca2+ release level was investigated. Moreover, they were cultured with a clonal human PDLSC line (line 1–17 cells). The main crystal structures of Na-WMTA were identical to the structures of D-WMTA. Globular aggregates with polygonal and needle-like crystals were found on D-WMTA and Na-WMTA, which included Ca, Si, Al, C and O. However, many amorphous structures were also identified on Na-WMTA. These structures consisted of Na and Cl, but did not include Ca. NaOCl immersion also reduced Ca2+ release level from whole WMTA discs. Line 1–17 cells cultured with D-WMTA formed many mineralized nodules and exhibited high expression levels of osteoblast-related genes. However, cells incubated with Na-WMTA generated a small number of nodules and showed low expression levels of osteoblast-related genes. These results indicated that NaOCl reduced Ca2+ release from WMTA by generating amorphous structures and changing its elemental distribution. NaOCl may also partially abolish the ability of WMTA to stimulate osteoblastic differentiation of PDLSCs.

We hereby report a case of advanced and recurrent colon cancer with long-term survival after 7 repeated surgical resections. A 73-year-old woman initially underwent right hemicolectomy and partial hepatectomy for an ascending colon cancer with synchronous liver metastasis. Pathological diagnosis of the tumors were moderately differentiated adenocarcinoma and metastasis to the liver compatibly. Final clinical stage was diagnosed as fT3N2M1(H1), fStage Ⅳ. But she was interrupted oxaliplatin-based adjuvant chemotherapy after 6 courses of CAPOX because of adverse drug reaction. One year after first operation, partial resection of right lung was performed for lung metastasis. Two years after first operation, 2nd resection of liver was performed for 2 liver metastatic lesions. Three years after first operation, 3rd partial liver resection, 2nd and 3rd partial lung resections were performed for metachronous metastases during 1 year. After 3 years recurrence free period, she complained of an induration of right neck and diagnosed as neck and supra clavicular lymph nodes metastases. Lymph nodes resection was performed. After the last operation, she has no sign of cancer recurrence for 1 year and 7 months, eventually she has been alive for 7 years and 7 months after the initial operation.

Osteomyelitis is a potentially devastating inflammatory bone disease that leads to bone destruction and loss. Treatment of osteomyelitis requires the removal of residual bacteria as well as osteogenesis with angiogenesis at the site of treatment. Use of an appropriate amount of copper (Cu) in treatment scaffolds may achieve these goals without the risk of toxicity. In this study, the surface of the carbonate apatite honeycomb scaffold was functionalized with Cu through a dissolution-precipitation reaction. The resulting scaffolds retained the honeycomb structure after immersion in CuCl

Introduction A post-marketing surveillance (PMS) study was conducted to confirm the long-term risk–benefit profile of sitagliptin administered to Japanese patients with type 2 diabetes mellitus (T2DM) under real-world conditions. Methods This prospective, multicentre, open-label PMS collected data from 3326 patients receiving sitagliptin according to the approved indication during the case registration period (July 2010–June 2012; observation period, 3 years). Safety was assessed via collection of data on adverse drug reactions (ADRs), estimated glomerular filtration rate (eGFR) and cardiovascular events whereas efficacy was assessed via changes in glycated hemoglobin (HbA1c). Results In 3265 patients evaluated for safety, 270 ADRs occurred in 207 (6.3%) patients overall. Metabolism and nutrition disorders were the most common class of ADRs, occurring in 58 patients overall (53 non-serious, 5 serious) with hypoglycaemia (17 patients, 0.52%) the most common ADR. In patients with eGFR > 90 mL/min/1.73 m2 at baseline (mean ± SD, 106.42 ± 18.11 mL/min/1.73 m2, n = 584), eGFR declined by 11.83 ± 17.53 mL/min/1.73 m2 (P

BACKGROUND/AIM Pancreaticoduodenectomy (PD) treatment outcomes in elderly patients have been reported to be acceptable, but the eligibility criteria are not clear. To elucidate the importance of PD in octogenarians in particular, we set five eligibility criteria for elderly patients based on preoperative cardiac and pulmonary function, nutritional status, daily activity status, and psychological independence status for the first time and evaluated prospectively whether the validity of patient selection was adaptable. PATIENTS AND METHODS The study population consisted of 222 patients with pancreaticobiliary cancer aged over 70 years. The patients were divided into two groups: 192 patients as septuagenarians and 30 as octogenarians. Postoperative morbidity and long-term outcome were compared between the two groups, and prognostic factors relating to survival time were identified. RESULTS Octogenarians had a significantly higher frequency of two or more comorbidities (p

Background To determine the prevalence of and risk factors for malignant transformation of ovarian endometrioma during dienogest therapy, which is very rare, we examined multiple cases of malignant transformation of ovarian endometrioma during dienogest therapy and performed a multivariate analysis of the records in our hospital. Methods The medical records of 174 patients who underwent DNGT for the treatment of OMA from June 1, 2011, to May 31, 2018, were reviewed retrospectively with the approval of the Human Ethical Committee of the University of Teikyo Hospital. And we provided one representative case of MT with obtaining written informed consent. To assess the effects of six representative factors, including advanced age, parity, surgical history, and endometrial cyst characteristics (including 3 factors), on the possibility of malignant transformation, we performed a multivariate logistic regression analysis. Results Of the 174 cases, 4 were diagnosed with malignant transformation, and these cases are reported. In the multivariate analysis, advanced age (P = 0.0064), nullipara (P = 0.0322), and enlargement (P = 0.0079) showed significant differences for malignant transformation occurrence. All 4 malignant transformation cases were among the 19 patients who had all of these 3 factors. Conclusions For a more accurate determination of the treatment approach, a larger sample size will be needed to determine the risk factors for malignant transformation during dienogest therapy.

Pancreaticoduodenectomy(PD)treatment outcomes in elderly patients have been reported to be acceptable, but the eligibility criteria are not clear. To elucidate the importance of PD in octogenarians, we set beforehand the eligibility criteria in the elderly and evaluated whether the validity of the patient selection was adaptable.The study population consisted of 244 patients aged70 years who hadpancreaticobiliary cancer. The patients were divided into 2 groups as follows: 32 patients in the octogenarian group and 212 patients in the septuagenarian group. The eligibility criteria were as follows: (1)cardiac function as ejection fraction of at least 40%, measured using Doppler echocardiography; (2)pulmonary function as forcedexpiratory volume in 1 second(FEV1.0%)of at least 50%on spirography; (3) nutritional status as serum albumin level of at least 3.0 g/dL; (4)daily activity status as Karnofsky performance status of at least 80%; and(5)psychological independence status as the capability of self-determination with respect to surgery. Postoperative morbidity and long-term outcome were comparedbetween the 2 groups, andprognostic factors relating to survival time were identified.The patients in the octogenarian group showed a significantly higher incidence rates of 2 or more comorbidities(p0.0001)andd elirium(p=0.024). The difference in mortality rate between the 2 groups was not significant. No significant difference in overall survival rate was found between the 2 groups(p=0.197). The independent prognostic factors relating to survival duration were intraoperative blood loss(p=0.0008)andd uration of surgery(p= 0.0091).The eligibility criteria for PD in elderly patients are also satisfactorily applicable to octogenarian patients. These criteria may be helpful when uncertainties arise regarding the selection of PD.

A 77-year-old woman with back and epigastric pains was diagnosed with pancreatic head cancer according to the result of contrast computed tomography, which showed a 25mm irregular low-density area at the pancreatic head infiltrating nearly half of the superior mesenteric plexus. There were no findings of lymph node metastasis or distant metastasis. The pretreatment diagnosis was borderline resectable(BR)pancreatic head cancer, cT3, N0, M0, cStage ⅡA. The patient was treated with gemcitabine plus nab-paclitaxel therapy. She developed Grade 3 neutropenia, and the dose was adjusted in order to continue chemotherapy. The size of the tumor had reduced to 15mm after 6 courses of the therapy, and the infiltration into the superior mesenteric plexus had also reduced. Therefore, the patient underwent subtotal stomach-preserving pancreatoduodenectomy and D2 lymph node dissection. The histopathological findings were invasive ductal carcinoma with R0 radical resection. The efficacy of preoperative adjuvant chemotherapy for BR pancreatic cancer has not been established yet, but improving the R0 resection rate with preoperative chemotherapy may contribute to an improvement in the outcome of pancreatic cancer.

Background and Aim: The recommendedind ication of transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC)is Barcelona Clinic Liver Cancer(BCLC)stage B HCC. But there are some cases in which we do not perform TACE because of liver damage with malnutrition in stage B. So we examined whether branched-chain amino acid (BCAA)improve nutritional status and perform TACE to contribute the prolongation of HCC survival.This study included8 8 patients treatedfor liver cirrhosis with HCC. All patients initially receivedBCAA granules. In patients with unchangedor decreasedAlb levels, BCAA granules were discontinuedandBCAA enrichednutrient was started. TACE for HCC were performedin those with an improvedChild -Pugh score.TACE were performedfollowing the aggressive intervention with BCAA nutritional education in 66 of 88(75%)patients. Finally, overall survival time was significantly extended in TACE group(p0.0001).Timely aggressive nutritional intervention in BCLC stage B HCC, early partial replacement with BCAA enrichednutrient before TACE may consequently contribute to improvement of the treatment outcome of HCC.

To diagnose postoperative pancreatic fistula(POPF)after pancreaticoduodenectomy(PD)during early post operative days, we evaluatedwhether a change in drain amylase value(DAV), WBC value, or CRP value predicted clinically significant POPF. One hundred thirty patients underwent substomach preserving PD(SSPPD)from 2005 to 2016 in our institution andwe examinedDAV, WBC, andCRP values at POD 1, 3, and 5. Seventy-five patients(57.7%)were equivalent to ISGPF grade A at POD 3.We analyzedthese 75 cases by following 5 factors(DAV at POD 31,500 U/L, DAV at POD 51,000 U/L,WBC value at POD 5≤9,000/mL, DAV of POD 5 decreased to 1/3 from DAV of POD 3, CRP value not improved≥50%from POD 3 to 5) andwere concernedwith the crisis of the grade B/C POPF at final diagnosis after POD 5. In conclusion, DAV of POD 5 decreased to 1/3 from DAV of POD 3, andthe CRP value having not improved(≥50%from POD 3 to 5)were significant risk factors for POPF grade B andC by Cox proportional hazard models.

Sonoporation is a promising method to intracellularly deliver synthetic gene carriers that have lower endocytotic uptake than viral carriers. Here, we applied sonoporation to deliver genes via polyethylene glycol (PEG)-grafted polymeric carriers that specifically respond to hyperactivated protein kinase A (PKA). PEG-grafted polymeric carrier/DNA polyplexes were not efficiently delivered into cells via the endocytotic pathway because of the hydrophilic PEG layer surrounding the polyplexes. However, the delivery of polyplexes into cells was significantly increased by sonoporation. The delivered polyplexes exhibited PKA-responsive transgene expression in PKA-overexpressing cells, but not in cells with low PKA activation. These results show that the sonoporation-mediated delivery of PEG-modified PKA-responsive polyplexes is a promising approach for safely applying gene therapy to abnormal cells with hyperactivated PKA.

The aim of this study was to identify predictive factors for emergent surgical interventions in patients with ovarian endometriomas hospitalized for pelvic inflammatory disease (PID).We retrospectively identified 22 patients with ovarian endometriomas who were hospitalized to manage PID. Patients were divided into two groups: those who required emergent surgery after failed antibiotic therapy (emergent surgery group, n = 6) and those who were discharged with successful antibiotic therapy (conservative group, n = 16). Logistic regression analysis was performed to analyze the factors associated with emergent surgery.Patients in the emergent surgery group were significantly more likely to have intrauterine or intrapelvic procedures before the onset of PID compared to those in the conservative group (83% vs 38%, odds ratio 8.33, 95% confidence interval 1.02-181.3; P = 0.048). The mean interval between the onset of PID symptoms and the commencement of parenteral antibiotic therapy was significantly longer in the emergent surgery compared to the conservative group (6.5 vs 1 day, odds ratio 1.28, 95% confidence interval 1.01-1.75; P = 0.041).A history of an intrauterine or intrapelvic procedure before the onset of PID was more likely to result in emergent surgery. A longer interval between the onset of PID symptoms and the commencement of parenteral antibiotics was also associated with emergent surgery. These findings could help to identify patients with ovarian endometriomas hospitalized with PID at higher risk of emergent surgical intervention.

BACKGROUND In decompensated liver cirrhosis, hypoalbuminemia still persists even after they have been treated with branched-chain amino acid (BCAA) granules. We prospectively evaluated whether BCAA enriched nutrient switched from BCAA granules would increase the serum albumin level, and consequently extend the survival time after hepatocellular carcinoma (HCC) treatment. METHODS This study included 77 patients treated for liver cirrhosis with HCC. After the nutritional assessment, all patients initially received BCAA granules. In patients with unchanged or decreased serum albumin levels, BCAA granules were discontinued and BCAA enriched nutrient was started. Transcatheter arterial chemembolization (TACE) for HCC were performed in those with an improved Child-Pugh score. RESULTS TACE were performed following the aggressive intervention with BCAA nutritional education in 54 of 77 (70.1%) patients. Finally, survival time was significantly extended in the TACE group (P

In general, transcatheter arterial chemoembolization(TACE)can obtain a high therapeutic effect on hypervascular tumors, but the definition of"hypervascular"is unclear. In this study, stainedtumor images on enhancedcomputedtomography( CT)were assessedaccord ing to CT-attenuation value. We investigatedwhether it is possible to estimate the treatment effect(TE)of TACE for hepatocellular carcinomas(HCCs).We studied 50 patients with unresectable HCCs who underwent TACE. A total of 141 tumors were diagnosed as HCC on enhanced CT. We measured andcalculatedthe ratios of CT-attenuation values of HCCs in the arterial phase to normal enhancedliver parenchyma in the portal phase(CT value ratio). We then evaluatedTE on each target nodule by enhancedCT, to examine the correlation between TE andthe CT value ratio.The CT-attenuation values were 119(range 61-180)hounsfieldunits(HU)for HCC and8 3(49-141)HU for liver parenchyma, andthe CT value ratio was 1.47(0.7-2.6). TE was positively correlatedwith the CT value ratio(p=0.0005). The cut-off value that suggestedfavorable results for TACE was 1.7 by receiver operating characteristic(ROC)analysis.The CT value ratio is useful for recognition of hypervascular tumors. We obtained favorable results in cases with a CT value ratio of 1.7 or more.

Hematopoietic stem cells form blood cells in bone marrow and reside in niches. Artificial environments that conserve these niches may generate bone marrow. Osteogenesis, angiogenesis, and material resorption must be regulated to create these environments. These processes are controlled by material composition and macro- and microporous structures. Here, three blocks with different micropore structures are fabricated. Carbonate apatite has nearly the same composition as natural human bone and their honeycomb structure facilitates cell penetration and survival. In samples with high microporosity, endosteum-like tissues such as sinusoids form in areas of material resorption and high local calcium concentration. These conditions resemble environments conducive to niche maintenance. Bone marrow-like tissues and megakaryocytes are successfully generated in this environment. Micropore structure is the most critical factor in bone marrow formation; however, the influences of material composition and macropore structure must also be considered. The results of this study may help develop treatments for bone marrow-related diseases and elucidate the components and functions of the hematopoietic stem cell niche.

We examined in vitro performance of the branched polyethylenimine (bPEI)-based gene carriers which respond to cancer-specific activation of protein kinase Cα (PKCα) to express plasmid DNA. The carriers were synthesized straightforward by using amide bond formation between a peptide terminal carboxyl and a primary amine group of bPEI. To examine the effect of the peptide contents in the carrier, we prepared several carriers with various peptide contents. The obtained polymers form polyplexes with tighter condensation of plasmid DNA than our previous gene carriers. After internalization of the polyplexes via endocytosis, the polyplexes effectively escaped from the endosome into cytosol. Then, the polyplexes showed a clear-cut response to PKCα to release plasmid DNA for gene expression. We determined the optimum contents of the peptides in carriers as 5 mol% to achieve the clear-cut response to PKCα.

We previously reported that of the various polyamidoamine (PAMAM) STARBURST dendrimer (generation 3, G3) (dendrimer) conjugates with cyclodextrins (CyDs), the dendrimer (G3) conjugate with α-CyD having an average degree of substitution of 2.4 (α-CDE (G3)) has the greatest potential for a novel carrier for siRNA in vitro and in vivo. To improve the siRNA transfer activity and the lack of target specificity of α-CDE (G3), we prepared folate-polyethylene glycol (PEG)-appended α-CDEs (G3) (Fol-PαCs) with various degrees of substitution of folate (DSF) and evaluated their siRNA transfer activity to folate receptor (FR)-overexpressing cancer cells in vitro and in vivo. Of the three Fol-PαCs (G3, DSF 2, 4 and 7), Fol-PαC (G3, DSF 4) had the highest siRNA transfer activity in KB cells (FR-positive). Fol-PαC (G3, DSF 4) was endocytosed into KB cells through FR. No cytotoxicity of the siRNA complex with Fol-PαC (G3, DSF 4) was observed in KB cells (FR-positive) or A549 cells (FR-negative) up to the charge ratio of 100/1 (carrier/siRNA). In addition, the siRNA complex with Fol-PαC (G3, DSF 4) showed neither interferon response nor inflammatory response. Importantly, the siRNA complex with Fol-PαC (G3, DSF 4) tended to show the in vivo RNAi effects after intratumoral injection and intravenous injection in tumor cells-bearing mice. The FITC-labeled siRNA and TRITC-labeled Fol-PαC (G3, DSF 4) were actually accumulated in tumor tissues after intravenous injection in the mice. In conclusion, the present results suggest that Fol-PαC (G3, DSF 4) could potentially be used as a FR-overexpressing cancer cell-selective siRNA delivery carrier in vitro and in vivo.

Recently, our group has proposed a novel gene-regulation system responding to cAMP-dependent protein kinase (PKA) that has been applied to living cells. In this study, human liver-specific bionanocapsules (BNCs) are used as a gene-delivery system to increase transfection efficiency and to target specific cell types. BNCs can efficiently deliver a target gene to human hepatocytes and hepatoma cells in vitro or in vivo. The combination of a signal-responsive gene-delivery system with BNCs led to an increase in the transfection efficiency and selectivity for hepatoma cells. Expression from the delivered gene was identified from PKA-activated hepatoma cells (HepG2), but not from colon tumor cells (WiDr). These results show that the combination of a gene-regulation system responding to an intracellular signal with BNC can be used for the selective treatment of human hepatoma cells.

Background Control of inflammation is essential for the clinical management of many common human diseases. However, there are few generally applicable strategies to convert an abnormal intracellular signal into a gene expression that leads to normalization of the intracellular environment. Recently, we proposed a novel strategy termed D-RECS (i.e. drug or gene delivery system responding to cellular signals) to convert an intracellular signal to transgene expression. In the present study, we applied this concept to inflammatory cells using Iκ-B kinase as a signal molecule that triggers the gene expression. Methods Candidate cationic substrates of Iκ-B kinase (IKK)β were synthesized and their reactivity was investigated. Then, polymers grafted with these peptides were prepared by radical polymerization. Polymer/DNA complexes (polyplexes) were prepared by mixing plasmid DNAs with the polymers. The behaviour of these polyplexes by adding IKKβ was examined. Furthermore, changes of gene expression were evaluated after the microinjection of polyplex into living cells under conditions of nuclear factor (NF)-κB activation. Results Synthetic peptides with additional lysine residues were well phosphorylated by IKKβ. Both the polymer and the polyplex were also phosphorylated by IKKβ. The results of gel shift assay showed that the polyplex was disintegrated and free DNA was released in the presence of IKKβ. The polyplex comprising-green fluorescent protein plasmid DNA and the polymer expressed the transgene in living cells exposed to a pro-inflammatory stimulus. Conclusions Our concept of cell-specific gene expression was demonstrated to work in inflammatory cells. This method may provide a unique strategy for gene therapy exclusively in inflammatory cells. Copyright © 2009 John Wiley & Sons, Ltd.

The molecular basis of the metastasis of hepatocellular carcinoma (HCC) is not fully understood. The aim of this study was to elucidate the crucial genes involved in metastasis of HCC.We compared expression profiles among highly metastatic HCC cell lines and non-metastatic HCC cell lines by using oligonucleotide array to identify genes associated with metastasis. We further investigated the effect of identified gene on cell motility and metastasis in vitro and in vivo. Finally, we examined immunohistochemistry in human tissue samples.We identified 39 genes whose expression levels were significantly correlated with metastatic ability (P0.05). Of these genes, we further investigated cortactin, because this cortical actin-associated protein is a substrate of Src, whose activation has been shown to be involved in HCC cell migration and metastasis. Overexpression of cortactin in a non-metastatic HCC cell line increased cell motility, and resulted in metastasis in an orthotopic model. Furthermore, immunohistochemical expression of cortactin revealed its significant overexpression in HCC with intrahepatic metastasis compared with HCC without intrahepatic metastasis (P0.005).Overexpression of cortactin may play a role in the metastasis of HCC by influencing cell motility, and cortactin could be a sensitive marker for HCC with intrahepatic metastasis.

We evaluated the clinical usefulness and safety of transradial approach for transcatheter arterial chemoembolization (TACE) in patients with hepatocellular carcinoma (HCC) compared with that of conventional transfemoral approach. The two groups (radial group, n = 177; femoral group, n = 150) of cases were retrospectively compared with regard to the successful rate of angiography or TACE, time required for catheterizaiton and complications. Hepatic angiography and TACE were completed in 174 (98.3%) of 177 cases in the radial group. There was no intergroup difference of time required for catheterization. Minor complications (dull pain, numbness) occurred in 8 (4.6%) patients in the radial group, and there were lower complications in the radial group compared to the femoral group. TACE by our new transradial approach was found to have therapeutic efficacy with lower complications comparable to that of the conventional transfemoral approach.

Deregulation of Wnt/beta-catenin signaling is thought to play a critical role in human carcinogenesis. Nemo-like kinase (NLK) is an evolutionarily conserved serine/threonine kinase that suppresses beta-catenin/T-cell factor (TCF) complex transcriptional activity through phosphorylation of TCF. Since NLK may be a tumor suppressor as a negative regulator of Wnt/beta-catenin pathway, we established tetracycline-inducible NLK and its kinase-negative mutant expression in DLD-1 human colon cancer cells to analyze the effect of NLK on cell growth and viability. The induction of wild-type NLK in DLD-1 cells caused suppression of cell growth whereas the kinase-negative mutant did not. Flow cytometry indicated that NLK expression increased the number of apoptotic cells but did not induce obvious cell cycle arrest. Apoptosis induction by wild-type NLK was confirmed using TUNEL assays. Our results suggest that overexpression of NLK may have targets other than TCF for induction of apoptosis in human colon carcinoma cells.

To determine the surgical intervention time, which is most likely to achieve a high success rate for blowout fracture repair without implants and the usefulness of treatment with an intramaxillary sinus balloon.Two hundred patients with isolated fractures of the orbit were evaluated by the Hess screen test, the Hertel exophthalmometer, and coronal computed tomography of the orbit. Operative criteria included diplopia within 30 degrees and enophthalmos3 mm. An inferior lid incision approach was used to expose the orbital floor for realignment of bone fragments. Eighty of the patients received a gingival incision, followed by an osteotomy to create a 10-mm opening into the maxillary sinus for placement of a silicon-Teflon-silicon balloon.The highest success rate, with diplopia completely improved in 66% of the patients, was observed when surgery was performed within 3 days after the injury. This success rate declined as surgical intervention was delayed. In 197 cases, enophthalmos was improved to2 mm postoperatively for patients who had surgery within 14 days. The balloon treatment was well tolerated and caused no complications.Surgery within 3 days is recommended in cases with diplopia and enophthalmos. An intramaxillary sinus balloon treatment was useful for the cases with large orbital floor fracture that could cause latent enophthalmos.

We report herein the usefulness of interventional radiological treatment( IVR) for hepatocellular carcinomas( HCCs), based on the results of transcatheter arterial chemoembolization( TACE) and transarterial infusion( TAI).The study included 256 cases of HCC. TACE and TAI were performed for durations permitted by the degree of liver damage. Results(: 1) TACE was performed in 224 cases( average: 4.5 times, range: 1-14 times), and TAI was performed in 32 cases( average: 2.3 times, range: 1-8 times).( 2) The 3- and 5-year survival rates for all cases were 45.5% and 31.6%, respectively.( 3) We classified all cases according to the number of HCCs, solitary, 2-4, and multiple HCCs, and found no significant differences in the survival rate between the 3 groups( p=0.207),( 4) TAI was followed by TACE in non-responsive cases, and the median survival time of the TAI group was 8.5 months.We can expect benefits from repeated TACE treatment in the multiple HCCs group, compared to the solitary HCC group. TAI followed by TACE might improve the prognosis of unresectable and recurrent HCCs. Therefore, we conclude that IVR has clinical benefit as local treatment for HCC.

PURPOSE: The aim of this study was to evaluate the role of histopathology of biopsy specimens in predicting depth of infiltration in early colorectal carcinomas before treatment. METHODS: Early colorectal carcinomas that had been resected surgically or endoscopically between 1984 and 1995 were analyzed. Histopathologic findings, including differentiation of adenocarcinoma and a desmoplastic response were investigated. RESULTS: One hundred nine early colorectal carcinomas consisted of 73 lesions of carcinomain situ, 13 submucosal carcinomas with minimum invasion, 8 lesions with moderate invasion, and 15 lesions with deep invasion. Of 73 carcinomain situ lesions, 72 (approximately 99 percent) showed well-differentiated adenocarcinomas and no desmoplastic response. Twelve (92 percent) of 13 submucosal carcinomas with minimum invasion also revealed well-differentiated adenocarcinoma without a desmoplastic response. Sixty-three percent (5/8)of lesions with moderate invasion revealed well-differentiated adenocarcinoma. None of the lesions had a desmoplastic response. Among lesions with deep invasion, 73 percent (11/15) demonstrated moderately differentiated adenocarcinoma, and 11 lesions had a prominent desmoplastic response (73 percent;P

A 49-year-old female was admitted with chief complaint of fecaluria on March 4th 1993. A radiation therapy had been performed for uterocervical cancer 18 years ago. The small intestine and bladder was detected by DIP- and cystogram simultaneously. It was diagnosed as an ileovesical fistula. A segmental resection of the ileum with partial cystectomy was performed on March 23rd. Histopathologically, the ileum showed a radiation enteritis. Eventually, we diagnosed that this ileovesical fistula was caused by radiation. After operation, an incompletion of suture occurred. So we made an ileostomy secondarily and performed hyperbaric oxygen therapy. The patient was getting well temporarily but died of gastric hemorrhage on May 1st.

Hemorrhagic cystitis resulting from radiation to pelvic visceral malignant lesions often might be incurable and there have been established no definitive treatment. We experienced 2 cases with radiation-induced severe hemorrhagic cystitis refractory to conventional therapy. The treatment with hyperbaric oxygen to control hematuria was performed and obtained successful results. Gross hematuria was disappeared and cystoscopic figure was remarkably improved. No remarkable side-effect was observed in both patient. This experience suggested that hyperbaric oxygen could be considered the primary treatment for patient with radiation-induced hemorrhagic cystitis instead of usual treatment.

Purpose To evaluate the reliability of a screening method in patients with common bile duct (CBD) stones before laparoscopic cholecystectomy (LC) based on predictive factors, and to determine the cases for which preoperative therapeutic endoscopic retrograde cholangiography (ERC) is indicated. Materials and methods Alkaline phosphatase, total bilirubin, amylase, and dilation of CBD results were defined as predictive factors for CBD stones. In cases in which all these 4 factors were negative, preoperative ERC was omitted, and in cases in which 1 of the 4 factors was positive, ERC was performed before LC. Results A total of 323 patients (83.7%) in whom all 4 factors were negative, ERC was omitted. In all the 63 patients (16.3%) in whom 1 of the 4 factors was positive, ERC was performed. In 52 (82.5%) of them, CBD stones were present and in 43 of them, LC was performed after endoscopic choledocholithotomy. Conclusions For cases in which any 1 of the 4 factors was positive before LC, it was strongly recommended to perform preoperative ERC.

Transcatheter arterial chemoembolization (TACE) was performed as the initial therapy for advanced hepatocellular carcinoma (HCC). However, no effective chemotherapy has been established for patients who did not respond to TACE, and for those the therapy was not suitable. Since 2004, transcatheter arterial infusion (TAI) chemotherapy using fine-powder cisplatin has been applied at our department to such cases mentioned before. This report described the therapeutic results of TAI therapy and presented a case of HCC accompanied by portal vein tumor thrombus (PVTT) for which the therapy was effective.TAI was performed using cisplatin in 16 patients with inoperable advanced HCC between 2004 and 2010. Cisplatin was infused into the proper hepatic artery at a dose of 65 mg/m2 for 30 minutes.TAI was performed in each patient 1.8 times on average, ranging from 1 to 8 times. Therapeutic Effect 4 (TE4) was obtained in 1 case, TE3 in 2, TE2 in 1, and TE1 in 11 cases. The only severe adverse event rated as grade 3 or above during the study were nausea and anorexia reported in 6.3% of the patients, and adverse events rated as less than grade 2 were increased serum hepato-biliary enzyme levels in 68.8%, nausea in 68.8% of the patients. The overall 1-year survival rate was 31.2%, and the 50% survival period was 314 days.As a treatment option following the initial TACE therapy for advanced HCC, TAI using cisplatin can improve the prognosis of HCC.

We report two cases of advanced hepatocellular carcinoma( HCC) with a portal vein tumor thrombus (PVTT) responding to transcatheter arterial infusion chemotherapy (TAI) using cisplatin (CDDP). Case 1: A 65-year-old male patient was diagnosed with unresectable multiple HCCs. After 5 courses of transcatheter arterial chemoembolization (TACE), the tumor became markedly enlarged, and a PVTT (Vp2) and lymph node metastasis were noted. TAI was conducted as the second- line chemotherapy, and the liver tumor, PVTT, and lymph node metastasis shrank. The clinical response was rated as a partial response (PR). Case 2: A 72-year-old female patient was diagnosed with single HCC. After a course of TACE, the tumor became markedly enlarged and was accompanied by a PVTT (Vp4). TAI was performed as the second-line chemotherapy, and the liver tumor and PVTT significantly shrank. The clinical response was also rated as a PR. TAI using CDDP as the second-line chemotherapy for advanced HCC is a promising treatment likely to contribute to the improvement of the patient's prognosis.

We present the results of classification of magnetic resonance cholangiography (MRC) focusing upon visualization of cystic duct for preoperative laparoscopic cholecystectomy (LC), along with assessments of a possible correlation between MRC classification and feasibility of LC.The study included a total of 663 patients who were diagnosed as cholelithiasis. MRC were classified into four types: Type A; entire biliary tract was visualized (+), Type B; cystic duct (+) but gallbladder was not(-), Type C; gallbladder (+), cystic duct (-), and Type D; gallbladder (-), cystic duct (-). The feasibility of LC was assessed using the following two variables: 1) percentages of cases in which LC was changed to open cholecystectomy (OC) and 2) operating time. The results were compared in respect of the relationship with the MRC classification.MRC were classified as follows: Type A (58.4%), Type B (15.2%), Type C (19.3%) and Type D (7.1%). A total of 124 patients (18.7%) had chronic cholecystitis, and a strong correlation with MRC type was noted (p0.0001). In 46 patients (7.5%), the surgical procedure was changed from LC to OC. The percentages of cases in which LC was changed to OC was significantly higher for Type C and Type D (p0.0001), so these two groups demonstrated prolonged operating time.Our MRC classification based on the visualization of cystic duct can reflect the feasibility of LC, and LC may be less feasible tin Type C, and in particular Type D patients.

Non-curative resection for cholangiocarcinoma usually leads to a poor prognosis. We present a case successfully treated with gemcitabine for residual tumor after extra hepatic bile duct resection with positive surgical margin. The patient was a 77-year-old male who was diagnosed as having a common bile duct cancer. Extra hepatic bile duct resection was performed, but intraoperative pathology diagnosed the surgical margin of duodenal-side bile duct was positive for cancer. Although additional resection of bile duct was done, pathological diagnosis resulted in positive margin again. We had to avoid pancreatoduodenectomy in light of the patient's wishes, so a curative resection could not be carried out. Adjuvant chemotherapy with gemcitabine(800 mg/m2 on days 1, 8 and 15 every 4 weeks)was started at the seventh postoperative day. A residual lesion was shown in the pancreas head by abdominal CT after 2 courses of chemotherapy. Follow-up CT was performed every 6 months, and the lesion gradually seemed to become unclear. Finally, CT showed disappearance of the residual lesion 28 months after surgery. The chemotherapy has been continued up to the present(3 years and 5 months after surgery). No evidence of recurrence nor adverse events of WHO grade 2 or more has been observed.

We evaluated a clinical usefulness and safety of transradial approach for transcatheter arterial chemoembolization (TAE) in patients with unresectable and recurrent hepatocellular carcinoma (HCC) compared with that of conventional transfemoral approach. The two groups (radial group; n=385, 1999.7-2007.3, femoral group; n=150, 1997.4-1999.6) of cases were retrospectively compared with regard to the successful rate of angiography, TAE, interventional time, Lipiodol retention pattern and complications. Hepatic angiography and TAE were completed in 379 (98.4%) of 385 cases in the radial group. There was no inter-group difference of interventional time. Minor complications (dull pain or numbness of puncture site) occurred in 29 (7.6%) patients in the radial group. Transradial approach has some advantages as follows: (1) No time will be required to stop breeding after removal of sheath. (2) The patient may freely walk after the examination. TAE by our new transradial approach for HCC was found to have a therapeutic efficacy with lower complications comparable to that of conventional transfemoral approach.

We present a case of interstitial pneumonitis induced by gemcitabine. The patient was a 48-year-old male who underwent pancreaticoduodenectomy with portal vein resection for ductal adenocarcinoma of the pancreas head. Twenty-two days after operation, adjuvant chemotherapy with gemcitabine(1,000 mg/m(2) on days 1, 8 and 15 every 4 weeks)was started. During three courses of chemotherapy, no adverse event of WHO grade 2 or more was observed. Seven days after the last infusion, fever and consciousness disorder emerged without respiratory symptoms. Arterial blood gas analysis revealed severe hypoxemia. Chest X-ray and CT showed diffuse bilateral interstitial infiltrates. Oxygenation and respiratory support were immediately given, and steroid pulse therapy with 1,000 mg/day of methylprednisolone was started. His symptoms and radiolographical findings were dramatically improved. The patient could be discharged on the 14th day after admission. Acute pulmonary toxicity induced by gemcitabine is rare, but could lead to severe complications. The review of reported cases showed the effectiveness of corticosteroid therapy for pulmonary toxicity due to the agent.

Transradial vascular intervention for heart disease has been expanding recently because it is less invasive, but this approach has only been used to treat abdominal disease at a few institutions. Since 2000, we have performed partial splenic embolization (PSE) via the transradial approach with the aim of achieving the least invasive procedure possible.During the past five years, transradial abdominal angiography was performed in 426 patients and eight of them (two men and six women undergoing PSE for LC) were included in this study. They ranged from 48 years to 72 years in age, consisting of three patients in Child class A and five in Child class B. To perform transradial partial splenic embolization, a 4-Fr sheath was inserted in the left radial artery. Under fluoroscopy, intrasplenic branches of the splenic artery supplying about 60-70% of the parenchyma of the spleen were selected and embolized with a newly developed transradial catheter inserted along a guide wire running through the descending aorta. After treatment, the puncture site was compressed by applying a tourniquet. No restriction on movement of the lower or upper extremities is required and the patient is allowed to walk freely.Transradial embolization was successful in all 8 patients. The procedure required 70 minutes on average. The mean platelet count was 3.3x10(4)/microL before treatment, and it increased to 16.3x10(4) and 11.3x10(4)/microL after two weeks and three months, respectively. All patients suffered from fever and pain after treatment, which were treated conservatively. Occlusion of the radial artery, digital paralysis, or splenic abscess did not occur in any patient.This transcatheter technique facilitates hemostasis and requires neither shaving of hair nor insertion of a urinary catheter. In addition, no restriction on movement is needed after the procedure. Consequently, this minimally invasive transradial approach is highly satisfactory for patients and should become a first-line approach for PSE.

Pseudoaneurysm of the cystic artery is rare; to our knowledge, fewer than 30 cases have been reported worldwide. We report the first case of an unruptured pseudoaneurysm of the cystic artery with concurrent acute calculous cholecystitis. We incidentally detected the aneurysm by contrast-enhanced computed tomography (CT) in the edematous, thickened wall of the gallbladder neck in a 71-year-old man, whereas in most of the reported cases the disease presented as hemobilia. Because of the high risk of aneurysm rupture in this location, we avoided such interventions as percutaneous cholecystostomy and laparoscopic cholecystectomy. The aneurysm was localized pathologically in the undisrupted gallbladder wall, and elective open cholecystectomy with ligation of the cystic artery was performed successfully. Our case highlights the usefulness of CT for both diagnosis and patient management. Open cholecystectomy with ligation of the cystic artery is demonstrated as a reasonable first line of treatment for this unusual condition.

The aim of this study was to determine useful predictive factors of common bile duct stones (CBDs) as diagnosed by endoscopic retrograde cholangiopancreatography (ERCP) in patients who underwent laparoscopic cholecystectomy (LC) for gallstones.A total of 510 patients underwent ERCP prior to LC. Also reviewed in each were clinical data, laboratory data, and ultrasonographic findings. Data were evaluated by uni- and multivariate analysis to determine which of the useful predictive factors thus far reported might be in the concurrence of CBDs.Univariate analysis identified jaundice, pancreatitis, ALT, total bilirubin, alkaline phosphatase, amylase, and CBD dilatation at ultrasonography as predictors. Multivariate analysis subsequently identified alkaline phosphatase (p0.0001), total bilirubin (p=0.0008), amylase (p=0.0009), and CBD dilatation at ultrasonography (p=0.0012) as independent predictive factors of CBDs. The estimates for the detection of CBDs, when the indication of ERCP is determined on the basis of the four predictive factors, were found to be as follows: sensitivity 97.6%, positive predictive value 78.6%, and positive accuracy 95.3%.It is advisable to ascertain by preoperative ERCP whether there might be any CBDs in patients about to undergo an LC for treatment of cholelithiasis insofar as the patient has one or more of these factors. It is concluded that an LC may be performed by omitting the prior ERCP, conversely, on patients devoid of all of these factors.

The prognosis of colorectal carcinoma (CRC) with liver metastasis varies from case to case. A standardized classification system for evaluation of the prognosis and the treatment is needed. Therefore, we developed a new staging system for CRC with liver metastasis (HM-stage) based on the survival data.We evaluated 148 CRC patients with liver metastasis treated between 1985 and 1999. Prognostic factors were identified based on a multivariate analysis. According to the final prognostic factors and hazard ratios, we defined the HM- stage.Three factors, including extent of liver metastasis, depth of tumor invasion and peritoneal metastasis, were identified to be the final prognostic factors. These factors were then assigned points. The patients were classified as being HM-stage I to IV by the sum total. The median survival time for each HM-stage were 37 months for HM-stage I, 23 months for II, 10 months for III and 7 months for IV respectively. A significant difference among each stage was recognized (p0.0001).This new staging system for CRC with liver metastasis is simple and should be clinically useful for both estimation of the prognosis and evaluation of the therapy in patients.

Dysadherin is a membrane glycoprotein expressed strongly in several human cancers. Overexpression of dysadherin in tumor cells is closely associated with malignant phenotype (e.g., metastasis) and poor prognosis. In our analysis, six pancreatic cancer cell lines showed a positive correlation between dysadherin expression and cell motility. Introduction of small interfering RNA (siRNA) against dysadherin into the Panc-1 cell line caused reduction of dysadherin expression and suppression of cell motility. In contrast, stable transfection of a dysadherin expression vector into the Capan-1 cell line increased cell motility. In vivo, the metastatic potential of orthotopically transplanted Capan-1 tumor cells in severe combined immunodeficient mice was increased by dysadherin overexpression. Cell morphology and actin organization were also influenced by modulation of dysadherin expression. Cells transfected with dysadherin siRNA tended to have a relatively larger, more spread shape and increased transverse actin stress fibers compared with parent cells and cells transfected with control siRNA. Our study suggests that dysadherin is able to modulate actin structures, stimulate cell motility, and contribute directly to the metastatic potential of human pancreatic cancer cells.

Cyclooxygenase-2 (COX-2) has been reported to play an important role in the pathogenesis of a variety of cancers. Little is known about the relationship between bile duct cancer associated with pancreaticobiliary maljunction (PBM) and COX-2.We compared the expression of COX-2 in bile duct epithelium in bile duct cancer with or without PBM.Immunohistochemical analysis showed that the rate of positive COX-2 expression was 42.9% for cancerous lesions and 7.1% for noncancerous lesions in cases of bile duct cancer without PBM. On the other hand, no positive expression of COX-2 was observed in cancerous lesion of bile duct cancer with PBM. Twenty % of the noncancerous bile duct mucosa with PBM was positive for COX-2.COX-2 is expressed in bile duct cancer while it is not expressed in bile duct cancer accompanied by PBM.

Of all of the epithelial ovarian cancers, clear cell carcinoma (CCC) of the ovary has the worst prognosis. We applied the oligonucleotide array technique to identify genes generally involved in CCC. Of the approximately 12,600 genes that were analyzed, 28 were expressed significantly differently between four CCC and seven non-CCC cell lines. Among 16 up-regulated genes in CCC, we further investigated a transcription factor, hepatocyte nuclear factor-1 beta (HNF-1 beta). We validated up-regulation of HNF-1 beta in CCC in terms of both mRNA and protein level using real-time quantitative reverse transcriptase-polymerase chain reaction and immunoblotting. Immunohistochemical analysis of 83 surgically resected ovarian cancers showed that almost all CCC specimens (21 of 22 cases) had nuclear staining for HNF-1 beta, whereas most non-CCC specimens (60 of 61 cases) showed no immunostaining or only focal and faint staining in the nucleus. Furthermore, we investigated the significance of HNF-1 beta expression in CCC using RNA interference. The reduction of HNF-1 beta expression by RNA interference induced apoptotic cell death in ovarian CCC cells, which was confirmed by terminal dUTP nick-end labeling and fluorescence-activated cell-sorting analyses. Our results suggest that HNF-1 beta is not only an excellent CCC-specific molecular marker but also a molecular target for therapy of ovarian CCC.

Amino acid (aa) 108-120 of L2 protein of human papillomavirus (HPV) type 16 contains a cross-neutralization epitope against genital HPV. We designed a placebo-controlled trial in healthy adults to evaluate the safety and immunogenicity of a synthetic peptide consisting of the aa 108-120 of HPV16 L2 (L2-108/120) region. A total of 13 volunteers were given nasal inoculations with 0.1 (n=5) or 0.5mg (n=5) doses of the peptides or placebo (n=3) without adjuvant at weeks 0, 4, and 12. Sera were collected before inoculation and at 6, 16 and 36 weeks. The inoculation caused no serious local and systemic complications. The inoculation generated anti-L2 antibodies binding to both HPV16 and 52 L1/L2-capsids in four of the five recipients in the 0.5mg group. Sera of the four recipients showed neutralizing activities against HPV16 and 52. Serological responses to the peptides were not found in the 0.1mg group and the placebo group recipients. This study suggests the L2-108/120 peptide is tolerable in humans and has the potential as a broad-spectrum prophylactic vaccine against genital HPV.