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Activation of c-MET through hepatocyte growth factor (HGF) increases tumorigenesis, induces resistance, and is associated with poor prognosis in various solid tumors. We previously reported that lung cancer cell lines had increased expression of c-MET due to gene amplification-induced cytotoxic drug resistance, and that resistant cells paracrine HGF and promote its resistance. However, the clinical significance of sHGF in patients with advanced or recurrent NSCLC, especially in patients treated with cytotoxic chemotherapy, is yet to be identified. Here, we present sHGF may be useful to predict tumor response and PFS in patients with advanced NSCLC. A total of 81 patients with 101 treatment regimens were investigated; 53 patients received first-line therapy and 48 patients received second-line therapy during the observation period. sHGF levels were evaluated using ELISA at 4 time-points: at pre-treatment, at response-evaluation (1-2 months after treatment initiation), at the best tumor response, and at the disease progression. As a control biomarker, serum carcinoembryonic antigen (CEA) was also evaluated. sHGF at response-evaluation in patients with progressive disease was higher compared with that of disease controls in both first-line and second-line treatment (median value (MV): under the limited of detection (LOD) vs. 0.40; P=0.0086 and MV: under the LOD vs. 0.41; P=0.0035, respectively). Positive-sHGF at response-evaluation predicts poor progression-free survival (PFS) compared with negative-sHGF in both first-line (median, 153.5 vs. 288.0; P=0.047) and second-line treatment (87.0 vs. 219.5; P=0.014). Lung adenocarcinoma subgroup analysis showed that in patients receiving second cytotoxic chemotherapy, there were no significant differences in PFS or HR between patients with low-CEA compared with those with high-CEA, but positive-sHGF at pre-treatment or at response-evaluation predicts poor PFS (35 vs. 132; P=0.0045, 50 vs. 215; P=0.0047, respectively). Simple and repeatable markers for activation of the HGF/c-MET pathway would provide rationale for future research investigating the merit of sHGF as a potential clinical biomarker to indicate administration of MET inhibitors. Citation Format: Takahiro Tsuji, Hiroaki Ozasa, Yuichi Sakamori, Takashi Nomizo, Tomoko Funazo, Yuto Yasuda, Hironori Yoshida, Hiroki Nagai, Young Hak Kim. Clinical impact of high serum hepatocyte growth factor in advanced non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5736. doi:10.1158/1538-7445.AM2017-5736

Osimertinib, a third-generation EGFR Tyrosine kinase inhibitor, has shown promising activity in EGFR mutation positive non-small cell lung cancer (NSCLC) which harboring T790M mutation. The mechanisms of resistance to osimertinib have been still unclear. To investigate the mechanisms of acquired resistance to osimertinib, we established the osimertinib-resistant cell line (PC9/osimertinib) from the NSCLC cell lines PC9, and measured the expression levels of ATP-binding cassette (ABC) transporters using quantitative real-time polymerase chain reaction. Expression profiling of ABC transporters revealed that the levels of ABCB1 gene expression were significantly increased in the PC9/osimertinib cells compared with the parental cells, but not other ABC transporters. Knockdown of ABCB1 expression using short hairpin RNA enhanced osimertinib cytotoxicity in PC9/osimertinib cells. These data suggest that increased expression of ABCB1 may have an important role in the acquired resistance to osimertinib. Citation Format: Takashi Nomizo, Hiroaki Ozasa, Takahiro Tsuji, Yuto Yasuda, Tomoko Funazo, Hironori Yoshida, Yuichi Sakamori, Hiroki Nagai, Young Hak Kim. Increased expression of ABCB1 could be associated with osimertinib resistance in non-small cell lung cancer cell line PC9 cell [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4161. doi:10.1158/1538-7445.AM2017-4161

Background Appropriate parental care by fathers greatly facilitates health in human family life. Much less is known from animal studies regarding the factors and neural circuitry that affect paternal behavior compared with those affecting maternal behavior. We recently reported that ICR mouse sires displayed maternal-like retrieval behavior when they were separated from pups and caged with their mates (co-housing) because the sires receive communicative interactions via ultrasonic and pheromone signals from the dams. We investigated the brain structures involved in regulating this activity by quantifying c-Fos-immunoreactive cells as neuronal activation markers in the neural pathway of male parental behavior. Results c-Fos expression in the medial preoptic area (mPOA) was significantly higher in sires that exhibited retrieval behavior (retrievers) than those with no such behavior (non-retrievers). Identical increased expression was found in the mPOA region in the retrievers stimulated by ultrasonic vocalizations or pheromones from their mates. Such increases in expression were not observed in the ventral tegmental area (VTA), nucleus accumbens (NAcc) or ventral palladium (VP). On the following day that we identified the families of the retrievers or non-retrievers, c-Fos expression in neuronal subsets in the mPOA, VTA, NAcc and VP was much higher in the retriever sires when they isolated together with their mates in new cages. This difference was not observed in the singly isolated retriever sires in new cages. The non-retriever sires did not display expression changes in the four brain regions that were assessed. Conclusion The mPOA neurons appeared to be activated by direct communicative interactions with mate dams, including ultrasonic vocalizations and pheromones. The mPOA-VTA-NAcc-VP neural circuit appears to be involved in paternal retrieval behavior.

We have reviewed 6 patients with acquired PRCA and investigated CD4/8 ratio in the bone marrow lymphocytes prior to treatment with cyclosporin A (CsA). CsA, as a remission induction therapy, produced CR in 3/6 patients, and PR+NR in 3/6 patients. CD4/8 ratio in CR group was high compared to PR+NR group. Moreover, the number of reticulocyte and hemoglobin concentration in CR group was high in comparison with PR+NR group. In acquired PRCA, CD4/8 ratio in the bone marrow lymphocytes may correlate with the disease status and the outcome of CsA treatment.

Learning Point for Clinicians Monoclonal gammopathy of renal significance (MGRS) is a recent and novel entity in monoclonal gammopathy of undetermined significance (MGUS). Crystalglobulinemia syndrome (CS), which commonly contributes to multiple myeloma (MM)-related kidney diseases, can occur in MGUS. Therefore, it is important for physicians to be aware of MGRS due to CS. An 86-year-old woman presented with fever, paresis and dysarthria. Although a vascular disorder was suspected in the central nervous system (CNS), magnetic resonance imaging of the head was normal. Her leukocyte count was 12.9 × 109/l, hemoglobin concentration 11.4 g/dl and platelet count 210.0 × 109/l. Blood chemistry was as follows: C-reactive protein 18.2 mg/dl, blood urea nitrogen 53.3 mg/dl, creatinine 2.4 mg/dl and calcium 10.3 mg/dl. Total serum protein was 6.4 g/dl with 2.9 g/dl albumin. Serum immunoglobulin (Ig) levels …

金沢大学疾患モデル総合研究センター, CD157, known as bone marrow stromal cell antigen-1, is a glycosylphosphatidylinositolanchored ADP-ribosyl cyclase that supports the survival and function of B-lymphocytes and hematopoietic or intestinal stem cells. Although CD157/Bst1 is a risk locus in Parkinson's disease (PD), little is known about the function of CD157 in the nervous system and contribution to PD progression. Here, we show that no apparent motor dysfunction was observed in young knockout (CD157-/-) male mice under less aging-related effects on behaviors. CD157-/- mice exhibited anxiety-related and depression-like behaviors compared with wild-type mice. These behaviors were rescued through treatment with anti-psychiatric drugs and oxytocin. CD157 was weakly expressed in the amygdala and c-Fos immunoreactivity in the amygdala was less evident in CD157-/- mice than in wild-type mice. These results demonstrate for the first time that CD157 plays a role as a neuro-regulator and suggest a potential role in pre-motor symptoms in PD. © 2014 Lopatina, Yoshihara, Nishimura, Zhong, Akther, Fakhrul, Liang, Higashida, Sumi, Furuhara, Inahata, Huang, Koizumi, Yokoyama, Tsuji, Petugina, Sumarokov, Salmina, Hashida, Kitao, Hori, Asano, Kitamura, Kozaka, Shiba, Zhong, Xie, Sato, Ishihara and Higashida., CC-BY 4.0

A 36-year-old woman was admitted to our hospital for further examination of an acute febrile illness with liver dysfunction. A peripheral blood smear displayed atypical lymphocytes. Cytomegalovirus (CMV) mononucleosis was diagnosed based on the detection of CMV-specific IgM and conventional CMV pp65 antigen. The physical examination on admission revealed signs of lower motor neuron right facial palsy. There were no significant cerebrospinal fluid findings, nor were there other neurological abnormalities. After receiving a short-course of oral corticosteroids, the patient gradually recovered from the facial paralysis. A one-month follow-up examination indicated that she had fully recovered neurologically, showing disappearance of CMV-DNA and a significant increase in the anti-CMV IgG titer. To our knowledge, there has been only one previous report describing CMV as the cause of an isolated facial palsy combined with CMV mononucleosis.

Nontuberculous mycobacterial (NTM) lung disease secondary to cystic fibrosis (CF) has been reported, but there is limited data about NTM prevalence in non-CF bronchiectasis. We retrospectively investigated the prevalence of NTM associated with diffuse panbronchiolitis (DPB), a disorder also characterized by reduced mucociliary clearance with bronchiectasis.We reviewed mycobacterial cultures, patient characteristics and computed tomography findings of 33 patients with DPB between January 2000 and December 2012. Prevalence was based on at least one positive NTM culture.Mean patient age was 51.5 years. During a mean 162.8-month follow-up, the prevalence of NTM in sputum was 21.2% (seven patients). Of the seven positive patients, six had Mycobacterium avium complex, one had M. kansasii and M. chelonae co-cultured with M. avium complex. Three patients were positive twice, and two had positive smears. The mean time from DPB diagnosis to the first positive result was 194.6 months. NTM-positive patients tended to have lower forced expiratory volume in 1 s (% predicted) than NTM-negative patients (50.0% vs 77.3%, P = 0.03), but there were no radiological or clinical differences between the two groups.Our observations suggest that NTM is found more often in DPB. Defects of mucociliary clearance may predispose individuals to NTM infection.

Hemophagocytic lymphohistiocytosis/hemophagocytic syndrome (HLH/HPS) is a reactive disorder of the mononuclear phagocytic system, characterized by generalized histiocytic proliferation with marked hemophagocytosis in the bone marrow [1]. It comprises two different categories: a primary (genetic) and a secondary (acquired) form. Malignant neoplasm-associated HPS (MAHS) is categorized as a secondary HLH. MAHS is mainly associated with lymphoma and rarely with other carcinomas [2]. To our knowledge, HLH associated with myelodysplastic syndromes (MDS) has not been described, although MDS with hemophagocytosis has been observed. Here, we describe a case of HLH associated with MDS, which presented with abundant CD8 T cells in the bone marrow and elevated plasma-soluble interleukin 2 receptor (sIL-2R). A 60-year-old Japanese man was admitted with pancytopenia, epitaxis, fever and general fatigue. A hematological examination showed a hemoglobin concentration of 6.5 g/dL, a platelet count of 14,000/lL and a leukocyte count of 3,200/lL with 42.8% atypical lymphocytes (Fig. 1a). Blood biochemistry showed increased levels of lactate dehydrogenase (LDH) (331 IU/L), ferritin (649 ng/mL) and sIL-2R (4,054 U/mL). NK cytolytic activity was measured by the standard 51-chromium (Cr) release assay. NK cytolytic activity was not reduced at both the E/T ratios of 10:1 (13.1%, normal range 8.9–29.5%) and 20:1 (22.5%, normal range 17.1–48.7%). Serological studies for hepatitis B, hepatitis C and HTLV-1 were negative. IgM antibodies to parvovirus were negative. Serological test for EBV revealed that anti-viral capsid antigen IgM was negative in the presence of anti-viral capsid antigen IgG (1609) and anti-EBV nuclear antigen antibody (409). The number of EBV DNA copies in serum was under 2.0 9 10 copies/mL (normal \200 copies/mL). Computed tomography (CT) revealed hepato-splenomegaly, ascites and bilateral pleural effusion. Bone marrow aspirate showed dyserythropoietic changes (abnormally lobulated nuclei) (Fig. 1c) and dysplastic megakaryocytes (micromegakaryocytes) (Fig. 1d). Erythrocytic precursors were 10.4% and bone marrow myeloblasts were 2.0% (Fig. 1e). Chromosome analysis showed severe complex karyotype: 54–57XY, ?1[2],?3[2],?4[4],?6[4],?8[4],add(9)(p22)[2],?11[3],add (15)(p11.2)[4],add(16)(q24)[4],add(19)(p13.1)[4],add(20) (p13)[4],?21[2],2–5mar[cp4]/46,XY[8]. A diagnosis of refractory anemia with excess blasts-1 (WHO classification) was made. In addition, bone marrow examination revealed increased numbers of atypical lymphocytes (Fig. 1b) and multiple sites of active hemophagocytosis (Fig. 2a–c). Atypical lymphocytes in bone marrow were positive for CD2, CD3 (surface, cytoplasmic), CD5, CD7, CD8, HLA-DR, TCR-ab, and negative for CD4. No monoclonal rearrangement of TCR beta, gamma and delta chain genes was detected by Southern blotting. These findings suggested that the atypical lymphocytes were reactive CD8 T cells. Combining fever, hemophagocytosis, splenomegaly, increased levels of ferritin and sIL-2R, a diagnosis of HLH was established. Although steroid therapy was administered for HLH, pancytopenia was not improved. Two months after the initial diagnosis, the levels of LDH and ferritin were further elevated (LDH 1,474 U/mL, ferritin 4,347 ng/mL). Bone marrow examination T. Tsuji (&) H. Yamasaki H. Tsuda Division of Hematology and Oncology, Kumamoto City Hospital, Kotoh 1-1-60, Kumamoto 862-8505, Japan e-mail: tsuji.takahiro@city.kumamoto.lg.jp

Photoluminescence (PL) properties of Eu-doped ZnO (ZnO:Eu) grown by a sputtering-assisted metalorganic chemical vapor deposition technique were investigated. In PL measurements at 300 K, the samples annealed at 600 °C for 30 min showed clear red-emission lines due to the intra-4f shell transition of 5 D 0 → 7 F J U=0-4) in Eu 3+ . In photoluminescence excitation (PLE) spectra, the PL was observed under the high-energy excitation above the band-gap energy of ZnO (indirect excitation) and the low-energy excitation resonant to the energy levels of 7 F 0 - 5 D 3 and 7 F 0 - 5 D 2 transitions in Eu 3+ (direct excitation). The PL lifetime under the indirect excitation was shorter than that under the direct excitations. These PL properties revealed that the energy transfer from ZnO host to Eu 3+ was accompanied under indirect excitation.

Photoluminescence (PL) properties of Er-doped β-FeSi 2 (β-FeSi 2 :Er) and Er-doped Si (Si:Er) grown by ion implantation were investigated. In PL measurements at 4.2 K, the β-FeSi 2 :Er showed the 1.54 μm PL due to the intra- 4 f shell transition of 4 I 13/2 → 4 I 15/2 in Er 3+ ions without a defect-related PL observed in Si:Er. In the dependence of the PL intensity on excitation photon flux density, the obtained optical excitation cross-section σ in β-FeSi 2 :Er ( σ =7×10 −17 cm 2 ) is smaller than that in Si:Er ( σ =1×10 -15 cm 2 ). In the time-resolved PL and the temperature dependence of the PL intensity, the 1.54 μm PL in β-FeSi 2 :Er showed a longer lifetime and larger activation energies for non-radiative recombination (NR) processes than Si:Er. These results revealed that NR centers induced by ion implantation damage were suppressed in β-FeSi 2 :Er, but the energy back transfer from Er 3+ to β-FeSi 2 was larger than Si:Er.

Background. To investigate the relation of estrogen and progesterone receptor (ER and PR) expression in endometrial cancer specimens to bone mineral density (BMD).Materials and methods. Subjects were 48 postmenopausal women with endometrial cancer treated with hysterectomy. Baseline characteristics included age, years since menopause (YSM), height, weight, and body mass index (BMI). Lumbar spine BMD (L2–4), the ratio of trunk fat to leg fat mass (trunk-leg fat ratio), body fat mass, and the percentage of body fat were measured by dual-energy x-ray absorptiometry (DEXA). ER and PR expression in endometrial cancer specimens were determined immunohistochemically.Results. Of 48 women, 32 (66.7%) were ER/PR-positive, nine (18.8%) were ER/PR-negative, three (6.3%) were ER-positive/PR-negative, and four (8.3%) were ER-negative/PR-positive. Lumbar spine BMD and trunk-leg fat ratio were significantly higher in women with ER-positive than in those with ER-negative (p

We developed a new pull-down assay for GTP-Rho and examined its level during cell cycle. HeLa cells were arrested in the S phase by thymidine and were enriched in the prometaphase, metaphase, telophase, and G(1) phase by collecting at 0, 45, 90, and 180 min after the release from the nocodazole arrest, respectively. The level of GTP-Rho did not change significantly from the S phase to the prometaphase, but increased thereafter, peaking in the telophase, and returned to the original level in the G(1) phase. The GDP-GTP exchange activity for Rho measured in cell lysates in parallel increased also during the mitosis with a peak in the metaphase. Using this system, we examined a role of ECT2, an exchanger for Rho GTPases, suggested to be involved in cytokinesis (Tatsumoto, T., Xie, X., Blumenthal, R., Okamoto, I., and Miki., T. (1999) J. Cell. Biol. , 147, 921-928). Expression of the dominant negative form of ECT2 completely suppressed both the rise of GTP-Rho in the telophase and the increased GDP-GTP exchange activity in the mitotic cell extracts. These results suggest a critical role of ECT2 in Rho activation during cytokinesis.

We retrospectively analyzed the proliferative activity and the centromeric copy number of chromosomes 8, 12, and 17 in both well-differentiated components and dedifferentiated ones of 10 dedifferentiated lip sarcoma cases by using MIB-1 immunostaining and fluorescence in situ hybridization (FISH) on the paraffin-embedded tissue specimens. The MIB1 labeling index (LI) in dedifferentiated components was higher than that in the well-differentiated components (p

Tyrosine kinase inhibitors (TKI) have been widely used for various malignancies. Off-target effects of these multi-kinase inhibitors, however, can have an adverse effect on the kidney.A 64-year-old...

Sclerosing stromal tumor (SST) is a rare ovarian neoplasm occurring predominantly in young women and is histologically characterized by cellular heterogeneity, prominent vasculature, and a pseudolobular appearance composed of cellular and hypocellular areas. In the current study, three cases of SST were ultrastructurally examined and the tumors were found to be composed of several kinds of cells, i.e., luteinized thecalike cells, spindle-shaped fibroblastic cells, and primitive mesenchymal cells. These findings thus supported the ovarian stromal origin of SST. Twelve cases of SST also were analyzed immunohistochemically and demonstrated an expression of vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) in the luteinized thecalike cells and its receptor, fms-like tyrosine kinase 1 (flt-1), in capillary to medium-sized blood vessels. Reverse transcription-polymerase chain reaction (RT-PCR) also showed an expression of VPF/VEGF messenger RNA in SSTs. Accordingly, the characteristic vasculature and edema of SSTs were considered to be associated with the expression of VPF/VEGF. In addition, a fluorescence in situ hybridization (FISH) analysis also showed cells with three copy number of chromosome 12 in 13-21% of all examined SST cells, which suggested the presence of chromosome 12 trisomy in SSTs as well as in other ovarian stromal tumors.

We retrospectively analyzed the proliferative activity and the centromeric copy number of chromosomes 8, 12, and 17 in three cases of fibrosarcoma and eight cases of cellular fibroma of the ovary using MIB-1 immunostaining, DNA flow cytometry, and fluorescence in situ hybridization (FISH) on paraffin-embedded tissue specimens. In our study, both the MIB-1 labeling index (LI) and the proliferative index (% of cells in S + G2 + M phase) in fibrosarcomas were higher than those in cellular fibromas. The FISH analysis demonstrated the sole abnormality of a gain of trisomy 12 cells in all eight cases of cellular fibroma. Both a gain of trisomy 12 cells and a gain of tetrasomy 12 cells were observed in one case of fibrosarcoma. A gain of trisomy 8 cells was observed in all two fibrosarcomas in which signals were detected. By contrast, neither a gain of trisomy 8 cells nor a gain of tetrasomy 12 cells was observed in any of the eight cases of cellular fibroma. Chromosome 17 showed disomy in all eleven cases. On the basis of these findings, a gain of trisomy 8 cells is therefore considered to be an adequately effective marker to distinguish between cellular fibroma and fibrosarcoma of the ovary, and it may also be related to the proliferative activity of fibrosarcoma of the ovary.

An 82-year-old female with a history of aortic dissection was admitted to our hospital for evaluation of thrombocytopenia and a bleeding tendency. Blood coagulation test data demonstrated that the patient had disseminated intravascular coagulation (DIC) secondary to aortic dissection. We initiated anti-fibrinolytic therapy with tranexamic acid. After this anti-fibrinolytic therapy, thrombocytopenia and the levels of fibrinogen and fibrinogen degradation products improved. In addition, we reviewed 7 other patients who were treated with tranexamic acid for DIC associated with aortic dissection. Anti-fibrinolytic therapy with tranexamic acid is effective for treating DIC secondary to aortic dissection.

症例は54歳の男性.突然の背部痛を主訴に受診.胸部X線写真で多発性の腫瘤陰影を指摘され,胸腔鏡下肺生検でlymphomatoid granulomatosis (LYG)と診断された. Cyclophosphamide (CPA) 50mg (隔日)とPrednisolone (PSL) 10mg (連日)の内服で治療を開始, 3カ月で腫瘤は著明に縮小し,その後CHOP療法を3コース追加して病変は消失した.丸二年を経過し無再発生存中である. LYGは希な疾患で,治療法が確立しておらず,治療方針を考える上で有意義な症例と思われた.

Upregulation of hepatocyte growth factor (HGF)/c-MET pathway causes drug resistance to anti-cancer agents such as epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors in non-small lung cancer (NSCLC) with EGFR mutation and cytotoxic agents in small cell lung cancer (SCLC), but the roles and mechanisms of c-MET in drug-resistant cancer cells are not clarified. We studied to reveal the role of c-MET overexpression in the resistant lung cancer cell lines and to demonstrate whether MET-inhibition could restore drug sensitivity in anti-cancer drug resistant cell lines via ABC transporters down-regulation. In this study we used the 7-ethyl-10-hydroxycamptothesin (SN-38)-resistant cell line (PC-6/SN-38) that was derived from the human SCLC cell line PC-6. We compared the expression levels of the purposed genes by quantitative real-time PCR (qRT-PCR) and western blotting (WB). MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) assay was used to measure cell viability of the resistant cells compared with the parental cells. Inhibition of c-MET activation was performed by c-MET inhibitors, PHA665752 and Crizotinib or small interfering RNA against c-MET. We found MET inhibitors (4 or 8μM) reduced cell growth and restored drug sensitivity (2μM) of resistant cells, PC-6/SN38, compared to parental cells. To reveal the mechanisms of c-MET in drug resistance we examined ATP-binding cassette (ABC) transporters expression in PC-6/SN-38 compared to the parental cells by qRT-PCR, and found the mRNA level of ABCG2, which effluxes SN-38 as substrate, in PC-6/SN-38 cells was extremely increased and another ABC transporters that do not efflux SN-38 were not. Recently we reported PC-6/SN-38 cells had c-MET overexpression, and c-MET inhibition in resistant cells such PC-6/SN-38 resulted in restoration drug resistance. Based on this report we examined c-MET inhibition by c-MET inhibitor and small interfering RNA against c-MET in PC-6/SN38, and qRT-PCR and WB were performed to investigate alteration of ABCG2 expression, and found both two c-MET inhibition reduced ABCG2 expression. In addition, to demonstrate c-MET upregulation in resistant cells is addicted to the ligand HGF, PC-6 cells were incubated with HGF treated for 3 weeks and made PC-6/HGF, then continued to incubate with HGF and SN-38 was added two times in a week. Although c-MET expression is equal in PC-6 and PC-6/HGF, after SN-38 treatment c-MET was overexpressed. Furthermore, serum HGF level in SCLC patients correlated with clinical course. We demonstrated upregulation of ABC transporters via HGF/c-MET signaling activation might be one of the mechanisms of acquired resistance to cytotoxic anticancer agents in lung cancer cells. Decreased ABC transporters expression through inhibition of c-MET activation might overcome drug resistance to cytotoxic agents. Citation Format: Yoshitaka Yagi, Hiroaki Ozasa, Takahiro Tsuji, Yuichi Sakamori, Takeshi Nomizo, Hiroki Nagai, Young Hak Kim, Ken Maeno, Tetsuya Oguri, Michiaki Mishima. Overexpression of ABC transporters through HGF/c-MET activation results in anti-cancer drug resistance in SCLC. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5045.

Background: Aldehyde dehydrogenase (ALDH) forms a superfamily of enzymes that catalyze conversion of aldehydes into carboxylic acids, and they are categorized into 19 families in human. Increased ALDH activity was reported to be a potential marker of cancer stem cell (CSC) in various solid tumors including lung cancer. Furthermore, High ALDH expression has been shown to be involved in drug resistance to conventional cytotoxic drugs. Recent studies revealed that ALDH7A1 is highly expressed in prostate cancer and associated with recurrence in patients with surgically resected non-small-cell lung cancer. However, there is not a fundamental link between ALDH7A1 expression and malignant phenotype, including drug resistance and colony formation. Material and methods: Anticancer drug resistant cell lines were established from lung cancer cell lines (PC-6, DMS53, PC-14, PC-9, NCI-H23 and NCI-H2228 cells) by exposure to various anticancer drug, including 7-ethyl-10-hydroxycamptothesin (SN-38), gemcitabine (GEM), cisplatin (CDDP), etoposide, paclitaxel (TXL), pemetrexed, amrubicin, erlotinib and crizotinib. Protein expression was determined by western blotting and gene expression was examined by RT-PCR. ALDH enzyme activity was measured by ALDEFLOUR TM assay. Proliferation was measured using MTS assay. ALDH7A1encoding plasmid was stably transfected into PC-14 cell. These transfected cells displayed sphere formation on ultralow binding plates and survived for more than 3 weeks. Results: We found that the levels of ALDH7A1 expression were higher in various anticancer drug-resistant lung cancer cell lines compared with the parental cells. ALDH enzyme activity was about 4-9 fold higher in SN-38, TXL, CDDP and GEM resistant cell lines (PC6/SN38, PC6/TXL, PC14/CDDP and PC14/GEM) than the parents cells. Up-regulation of ALDH7A1 expression by using a forced expression vector in PC14 cell altered cytotoxicity to SN-38 and TXL. Furthermore, overexpression of ALDH7A1in PC-14 cells enhanced sphere formation relative to the cells transfected with control vector. Conclusion: These results suggested that increased ALDH7A1 expression in lung cancer cell lines might be one of the mechanisms of acquired resistance to anticancer agents and enhance colony formation. Citation Format: Yuichi Sakamori, Hiroaki Ozasa, Eiji Kunii, Yoshitaka Yagi, Takahiro Tsuji, Takeshi Nomizo, Hiroki Nagai, Young Hak Kim, Ken Maeno, Tetsuya Oguri, Michiaki Mishima. Increased ALDH7A1 expression enhances the resistance to the anticancer drugs and colony formation in lung cancer cell lines. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2946.

Alectinib, a second-generation anaplastic lymphoma kinase (ALK) inhibitor, has shown promising activity in ALK-rearranged non-small cell lung cancer (NSCLC). The mechanisms of drug resistance to alectinib have been still unclear. To explore whether overexpression of ATP-binding cassette (ABC) transporters might provide a potential mechanism of alectinib resistance in lung cancer, we established alectinib-resistant cell lines from NCI-H2228 and primary ALK-rearranged cells and evaluated the expression of ABC transporters. We established ALK-rearranged NSCLC cell lines (KTOR 1, 2, and 3) from tumor cells in pleural effusion obtained from 3 patients with alectinib naïve ALK-rearranged NSCLC. In the 2 of 3 patients, tumor cells in pleural effusion were also obtained when disease progression was observed during alectinib treatment (KTOR1-RE and KTOR2-RE). Expression profiling of ABC transporters using quantitative real-time polymerase chain reaction (qRT-PCR) revealed that the expression of ABCB1 was significantly increased in KTOR1-RE and KTOR2-RE compared with the parental cells (4.6 and 10.8 folds), but not other ABC transporters. Then, alectinib-resistant NCI-H2228 cell lines (H2228-AR) were established by exposing the parental cells to stepwise-increasing alectinib up to 5μM for 10 months. We found that the ABCB1/MDR1 expression in H2228-AR cells were significantly increased (2.09-16.6 folds) compared with the parental cells using qRT-PCR and immunoblotting. Then, we examined the alternation of alectinib cytotoxicity by inhibition of ABCB1. Knockdown of ABCB1 expression using small interfering RNA and inhibition of ABCB1 by verapamil enhanced alectinib cytotoxicity in H2228-AR cells. Next, the levels of ABCB1 expression after short-term alectinib exposure were examined in NCI-H2228 and the three alectinib naïve ALK-rearranged NSCLC primary cell lines (KTOR 1, 2, and 3). In all the four ALK-rearranged cell lines, exposure to 100 nM alectinib for 72 hours induced increased expression of ABCB1 compared with control medium. These data suggested that increased expression of ABCB1/MDR1 might be, in part, responsible acquired resistance and sensitivity to alectinib in ALK-rearranged NSCLC cells. Citation Format: Takahiro Tsuji, Hiroaki Ozasa, Yuichi Sakamori, Takashi Nomizo, Yoshitaka Yagi, Hiroki Nagai, Young Hak Kim, Michiaki Mishima. Increased expression of ABCB1/MDR1 could be associated with alectinib resistance in ALK-rearranged lung cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2941.

子宮頸管ポリープに上皮内癌および微小浸潤扁平上皮癌を認めた各症例を経験したので報告する. 症例1は61歳. 人間ドックでの子宮頸部スメア異常を主訴として, 当科を受診. 外来受診時のスメアはclass IVであり, 径5mmの頸管ポリープを切除したところ, 組織学的に微小浸潤扁平上皮癌と診断された. 症例2は37歳. 集団検診にて子宮頸部スメア異常を指摘され当科を紹介初診となった. 初診時のスメアはclass III aであった. 径6mmの頸管ポリープ表面に白色上皮を認め, ポリープ切除を施行したところ, 組織学的に上皮内癌と診断された. 子宮頸管ポリープでは悪性の頻度は小さいが, ときに癌を合併することがあり, 頸管ポリープを切除した場合には組織学的診断を必ず行うことが必要であるといえる.

単純子宮全摘出術の4年後に腟断端に再発した子宮頸部悪性腺腫 (adenoma malignum) を報告する. 症例は41歳, 1回経妊1回経産婦で, 腟断端腫瘤の生検標本に腺癌が認められたため, 当科へ紹介され受診した. 初診時, 腟断端左側に数個の小豆大腫瘤を認め転移性腺癌を疑い, CTによる全身検索を施行したが, 原発巣と考えられる腫瘍は他に認められなかった. 4年前の摘出子宮標本を再鏡検したところ, 一部の頸管腺は既存の頸管腺領域を越えて小嚢胞を形成しており, その組織像は腟断端腫瘤の生検組織像ときわめて類似するものであった. 免疫組織化学的検索では, 腫瘍細胞の腺腔側の細胞膜上にCEA陽性所見が認められ, 4年前の子宮頸部標本をadenoma malignumと診断した.

Recurrence of non-Hodgkin's lymphoma more than 5 years after the initial diagnosis is rare. When late relapse occurs, it is difficult to determine whether it is a true recurrence or a new lesion. We experienced a case of an 81-year-old woman who developed central nervous system (CNS) lymphoma 12 years after remission of ocular adnexal lymphoma. Both showed the histology of diffuse large B-cell lymphoma. To elucidate whether the CNS lymphoma was clonally related to the first lymphoma, rearrangement of the immunoglobulin heavy chain genes of each lymphoma was studied using a polymerase chain reaction-based method. The results revealed that the sizes of the amplified products of the rearranged regions from the two lymphomas were different. This suggested different clonal origins of the lymphomas. It is clinically important to determine the origin of a second neoplasm because patients with a clonally related second lymphoma are usually treated with more intensive regimens, while those with a clonally unrelated lymphoma receive standard first-line therapy. The present case shows that, in the case of recurrent non-Hodgkin's lymphoma, not only histological confirmation but also genetic assessment is important to clarify the origin of the second lymphoma.

Forward reasoning engine is a computer program to automatically draw new conclusions by repeatedly applying inference rules. An automated forward reasoning engine for general-purpose, named FreeEnCal, has been proposed and developed. Then, to improving its performance and generality, fast algorithms and a general reasoning algorithm are proposed and implemented, but separately. Until now, there is no practical implementation of FreeEnCal that are adopted those algorithms. This paper shows the practical implementation of FreeEnCal and its practical usage. We can expect to use the practical FreeEnCal for an alternative tool to prove something, an enumeration tool by using forward deduction, a web service of automated forward reasoning.

A previously healthy 59-year-old woman was admitted to our hospital due to petechiae. Investigations showed profound thrombocytopenia (1.5×10(4)/µl) and mild elevation of transaminases. The serological examination revealed acute cytomegalovirus (CMV) infection. We intermittently measured CMV load in her clinical course. The petechiae improved with no therapy. One month later, the platelet count was increased up to 7.6×10(4)/µl and CMV-DNA was reduced to undetectable levels. CMV-induced thrombocytopenia in an immunocompetent adult is rare. We also recognized the association of platelet counts and virus load in the natural course of this patient with CMV-induced thrombocytopenia.

The incidence of iatrogenic parasitic uterine myomas associated with the use of a laparoscopic morcellator has been increasing over the past decade. Recently, we encountered a very rare case with a large parasitic myoma measuring 12 cm in diameter in the abdominal subcutaneous adipose tissue along an abdominal longitudinal surgical scar. The patient had twice undergone abdominal myomectomy for multiple fundal myomas. This is the first report describing a case with as large a parasitic myoma presenting in the suprafascial adipose tissue under the surgical scar after laparotomy. In such a case demonstrating a solid tumor of unknown cause after a gynecologic surgical procedure, a parasitic myoma must be included in the differential diagnosis.

Bone disease is a major symptom of multiple myeloma, which results from excessive osteoclast activation and impaired osteoblast function. Novel antimyeloma agents, such as immunomodulatory drugs and bortezomib, alter bone metabolism in patients with this disease. However, recent studies have shown that bortezomib affects bone formation and healing, but immunomodulatory drugs do not. We report a case of refractory multiple myeloma that was treated with lenalidomide combined with dexamethasone (RD regimen). As a result, the patient’s myeloma protein expression was markedly reduced, and her hematological profile was improved. However, the treatment also induced tetany by reducing her serum calcium and magnesium levels and elevating her alkaline phosphatase levels. In addition, conventional radiography and computed tomography detected osteosclerosis. These findings suggest that the RD regimen affects bone formation, possibly by activating osteoblasts. This is to our knowledge the first report to suggest that lenalidomide, an immunomodulatory drug, affects osteogenesis, at least in a certain subpopulation of patients with multiple myeloma.

The present study investigated the clinical outcome of stage Ia1 squamous cell carcinoma (SCC) of the uterine cervix at a single institute.Subjects were 84 patients with cervical SCC, International Federation of Gynecology and Obstetrics stage Ia1 diagnosed according to conization findings between January 2000 and June 2009. Correlations between clinical outcome and pathological findings on initial conization were investigated.Of 65 hysterectomy (without pelvic lymphadenectomy) patients, two (3.1%) showed metastasis to the vaginal wall, and two (3.1%) to the pelvic lymph nodes. Five patients (6.0%) showed lymphovascular invasion, and three (3.6%) showed confluent patterns of stromal invasion (confluent invasion) on initial conization findings. All of these five patients were included in the hysterectomy group. Of 19 patients with conization alone, two showed vaginal metastasis. Of four patients with vaginal metastasis, two showed lymphovascular invasion and one showed confluent invasion despite negative conization margin. In patients with nodal metastasis, stromal invasion of 3.0 mm and 2.8 mm, and horizontal spread of 6.8 mm and 6.9 mm were observed on conization, respectively. All patients are currently alive due to adequa\e therapy including chemo-radiation for six patients with recurrent disease.In patients showing stromal invasion of nearly 3.0 mm and horizontal spread of nearly 7.0 mm in stage Ia1 cervical SCC, pelvic lymphadenectomy may be considered. Even for patients showing a negative conization margin, careful follow up is necessary, because vaginal metastasis sometimes occurs.

Malignant fibrous histiocytoma (MFH) of the vulva is extremely rare; to date, there have been nine case reports. Almost all of these cases involved middle-aged women. We encountered a 21-year-old woman with a 4.5 × 2.6 cm superficial, localized, exophytic tumor of the right vulva. Microscopic findings on punch biopsy of the tumor initially suggested a vulvar sarcoma. The patient underwent wide local excision of the vulva. The extirpated specimen demonstrated the pleomorphic type of MFH with myxoid stroma of the vulva. The clinical stage was found to be IIB, based on the American Joint Committee on Cancer staging system. Chromosomal analysis of the tumor using the conventional G-band method was normal (46XX). This seems to be a very rare case of MFH of the vulva in a young woman. Physicians should therefore include MFH in the differential diagnosis of vulvar tumors, even though it is a rare disease.

Background The HIV-1 regulatory protein Rev, which is essential for viral replication, mediates the nuclear export of unspliced viral transcripts. Rev nuclear function requires active nucleocytoplasmic shuttling, and Rev nuclear import is mediated by the recognition of its Nuclear Localisation Signal (NLS) by multiple import factors, which include transportin and importin β. However, it remains unclear which nuclear import pathway(s) predominate in vivo, and the cellular environment that modulates Rev nucleocytoplasmic shuttling remains to be characterised. Results In our study, we have identified the cellular protein HIC (Human I-mfa domain-Containing protein) as a novel interactor of HIV-1 Rev. We demonstrate that HIC selectively interferes with Rev NLS interaction with importin β and impedes its nuclear import and function, but does not affect Rev nuclear import mediated by transportin. Hence, the molecular determinants mediating Rev-NLS recognition by importin β and transportin appear to be distinct. Furthermore, we have employed HIC and M9 M, a peptide specifically designed to inhibit the transportin-mediated nuclear import pathway, to characterise Rev nuclear import pathways within different cellular environments. Remarkably, we could show that in 293T, HeLa, COS7, Jurkat, U937, THP-1 and CEM cells, Rev nuclear import is cell type specific and alternatively mediated by transportin or importin β, in a mutually exclusive fashion. Conclusions Rev cytoplasmic sequestration by HIC may represent a novel mechanism for the control of Rev function. These studies highlight that the multivalent nature of the Rev NLS for different import receptors enables Rev to adapt its nuclear trafficking strategy.

To identify genes required for brain development, we previously performed in vivo RNA interference (RNAi) screening in Drosophila embryos. We identified pebble as a gene that disrupts development of the Drosophila nervous system. Although pebble has been shown to be involved in neuronal development of Drosophila in several screens, the involvement of Ect2, a mammalian ortholog of pebble, in mammalian neuronal development has not been addressed. To examine the role of Ect2 in neuronal differentiation, we performed Ect2 RNAi in the mouse neuroblastoma × rat glioma NG108-15 cell line. Depletion of Ect2 resulted in an increased proportion of binucleate cells and morphological differentiation of NG108-15 cells characterized by the outgrowth of neurites. These morphological changes were correlated with an increased level of acetylcholine esterase mRNA. In addition, expression of Ect2 was decreased in differentiated NG108-15 cells induced by dibutyryl cyclic AMP. These findings indicate that Ect2 negatively regulates the differentiation of NG108-15 cells and suggest that Ect2 may play a role in neuronal differentiation and brain development in vivo.

To investigate the relative contributions of weight-bearing and non-weight-bearing effects of adipose tissue to bone mineral density (BMD) in postmenopausal women. The subjects were 228 postmenopausal women aged 50-75 years. Age, years since menopause (YSM), height, body weight, and body mass index were recorded. Trunk fat mass, body fat mass, bilateral leg BMD and lean (muscle) mass were measured by whole body scanning with dual-energy X-ray absorptiometry. The relationships of BMD to trunk and body fat mass were investigated using uni- and multivariable analyses. The amount of trunk fat mass and body fat mass were 8.7 ± 3.6 kg and 19.0 ± 5.9 kg, respectively. On Pearson's correlation test, right leg BMD was positively correlated with trunk fat mass (r=0.268, P0.001) and body fat mass (0.299, P0.001). On multiple linear regression analysis, trunk fat mass (t-value = 3.500, P0.001), age (-2.431, P0.05), and YSM (-2.564, P0.01) were independent significant predictors of right leg BMD. However, body fat mass was not a predictor of BMD (-0.465, P=0.642). These relationships remained significant after further adjusting for right leg muscle mass. Trunk fat mass rather than body fat mass is a significant predictor of leg BMD at the most weight-bearing site, despite being less than half the amount of body fat mass. Thus, adipose tissue contributes more to BMD through non-weight-bearing effect rather than weight-bearing effect.

A 65-year-old female was admitted to our hospital for evaluation of transfusion-dependent progressive anemia. She had a history of rheumatoid arthritis for twenty-four years. Two years earlier, MDS (refractory anemia) was diagnosed based on laboratory findings and bone marrow examination. After diagnosis, the patient received anabolic steroid, Vitamin D3 and Vitamin K2. Although her hemoglobin level was maintained at 8.0 g/dl approximately 9.0 g/dl until January 2009, anemia gradually progressed thereafter. In April, we recognized marked anemia (Hb 6.0 g/dl) and reticulocytopenia (2.0 per thousand), but this was not accompanied by any other significant changes in laboratory findings. Bone marrow examination demonstrated a low percentage of erythroid precursors without an increase of blast cells. Rheumatoid arthritis remained stable by low dose steroid and NSAID administration. We did not recognize evidence suggesting any other cause of acquired PRCA, such as thymoma, human parvovirus B19 infection or drugs. A diagnosis of MDS with erythroid hypoplasia was made. The patient was successfully treated by an immunosuppressive regimen using cyclosporine. MDS with erythroid hypoplasia, coexisting with rheumatoid arthritis, is rare. To our knowledge, this is the third reported case.

Only limited data are available regarding mucin antigen expression in adenocarcinoma of the uterine cervix.Fifty-two patients with mucinous adenocarcinoma of the endocervical type were enrolled for making clear implication of the mucin antigens. The expression of mucin antigens in hysterectomized specimens were determined immunohistochemically. Clinicopathological characteristics and prognosis included International Federation of Gynecology and Obstetrics (FIGO) stage, histological grade, nodal and ovarian metastases, overall and disease-free survivals. The relationships of the expression of mucin antigens to various variables were investigated using uni- and multivariate analyses.The majority of mucinous adenocarcinoma showed overexpression of MUC1 and MUC16. Overexpression of both MUC1 and MUC16 was associated with lower survival rates. Particularly, overexpression of MUC1 was associated with a lower disease-free survival rate and lymph node metastasis. However, absence of expression of MUC1 and/or MUC16 was associated with longer overall and disease-free survival. In cases classified as FIGO stage Ib (n = 35), after adjusting for patient age at diagnosis and stratifying by histological grade, MUC1 and/or MUC 16 overexpression was an independent prognostic factor for overall survival (hazard ratio [HR] = 6.16, 95% confidence interval [CI] = 1.01-118.5, P0.05). After stratifying by ovarian metastasis, MUC1 and/or MUC 16 overexpression was also an independent prognostic factor for disease-free survival (HR = 5.50, 95% CI = 0.82-87.4, P0.05).The expressions of MUC1 and/or MUC16 may be available as independent prognostic factors for the endocervical type of mucinous adenocarcinoma.

Bacteroids are defined as the symbiotic forms of Rhizobium or Bradyrhizobium cells in the root nodules of their legume host. The differentiation to bacteroids involves various physiological changes and may be associated with some genetical changes. Single-colony isolates from a nodule formed by a Bradyrhizobium strain often differed in their effectiveness and intrinsic antibiotic resistance (Weaver and Wright 1987; Ozawa unpublished data). Detailed analysis of the changes in the gene structure requires the isolation of individual bacteroids from a nodule. However it has been reported that the viability of bacteroids in culture is very low and only a small fraction of a bacteroid population could produce colonies on yeast extract-mannitol (YEM) agar (Sutton et al. 1977). Bergersen (1974) concluded that the reversion of bacteroids to the vegetative, growing form is very rare.

Objective: The aim of this study was to assess fluorodeoxyglucose (FDG) uptake by leiomyomas in relation to histopathologic subtype and the menstrual state. Methods: We performed a retrospective study of 92 patients referred for uterine leiomyomas confirmed by histopathologic findings. On FDG-positron emission tomography, quantitative evaluation was performed using the mean values of the standardized uptake value (SUV) with reference to magnetic resonance imaging findings. Of the 92 patients, 56 had typical leiomyomas (41 premenopausal and 15 postmenopausal women), whereas 36 patients with subtype leiomyomas consisted of 21 premenopausal and 15 postmenopausal women. Results: Although no significant differences in SUV were observed between the typical and subtype leiomyomas, tumoral SUV was significantly higher in the premenopausal patients than in the postmenopausal patients. Uterine tumoral FDG uptake during the proliferative phase was also slightly lower than in the other menstrual phases. Conclusions: The FDG uptake by leiomyomas in premenopausal women was greater than that in postmenopausal women.

To clarify whether mutations in the large T gene encoded by Merkel cell polyomavirus affect the expression and function of large T antigen in Merkel cell carcinoma cases, we investigated the expression of large T antigen in vitro and in vivo. Immunohistochemistry using a rabbit polyclonal antibody revealed that large T antigen was expressed in the nuclei of Merkel cell carcinoma cells with Merkel cell polyomavirus infection. Deletion mutant analyses identified an Arg-Lys-Arg-Lys sequence (amino acids 277-280) as a nuclear localization signal in large T antigen. Sequence analyses revealed that there were no mutations in the nuclear localization signal in any of the eleven Merkel cell polyomavirus strains examined. Furthermore, stop codons were not observed in the upstream of the nuclear localization signal in any of the Merkel cell carcinoma cases examined. These data suggest that the nuclear localization signal is highly conserved and functional in Merkel cell carcinoma cases.

Merkel cell carcinoma is a rare malignancy that sometimes occurs in the skin of elderly people. Recently, a new human polyomavirus, Merkel cell polyomavirus (MCPyV) was identified in Merkel cell carcinoma. In the present study, MCPyV-DNA was detected in 6 of 11 (55%) cases of Merkel cell carcinoma by nested PCR and real-time PCR. Histologically, MCPyV-positive cases showed round and vesicular nuclei with a fine granular chromatin and small nucleoli, whereas MCPyV-negative cases showed polygonal nuclei with diffusely distributed chromatin. Real-time PCR analysis to detect the MCPyV gene revealed that viral copy numbers ranged 0.04–0.43 per cell in cases of Merkel cell carcinoma. MCPyV was also detected in 3 of 49 (6.1%) cases of Kaposi's sarcoma (KS), but not in 192 DNA samples of other diseases including 142 autopsy samples from 20 immunodeficient patients. The MCPyV copy number in KS was lower than that in Merkel cell carcinoma. PCR successfully amplified a full-length MCPyV genome from a case of KS. Sequence analysis revealed that the MCPyV isolated from KS had 98% homology to the previously reported MCPyV genomes. These data suggest that the prevalence of MCPyV is low in Japan, and is at least partly associated with the pathogenesis of Merkel cell carcinoma. J. Med. Virol. 81:1951–1958, 2009. © 2009 Wiley-Liss, Inc.

A 79-year-old man was admitted to our hospital due to generalized petechiae. Four days before onset, the patient had been administered an influenza vaccine. The platelet count, which was 13.6x10(4)/microl before vaccination, decreased to 0.4x10(4)/microl. Although he received various treatments, such as high-dose immunoglobulin therapy, prednisolone, and splenectomy, they were ineffective. Three weeks after CyA administration, the platelet count began to increase. Eighty days after administration, it reached 3.0-5.0x10(4)/microl. Thrombocytopenia after influenza vaccination is rare, and no refractory case has been previously reported.

An 88-year-old female was admitted to our hospital due to a right breast tumor and biclonal gammopathy (IgG-lambda and IgA-lambda). CT scan showed systemic lymphadenopathy. Tumor specimens were comprised of medium-sized atypical lymphocytes. Lymphoepithelial lesions and plasmacytic differentiation were observed. The atypical lymphocytes were positive for CD20, CD79a, Bcl-2, CD5 and negative for CD10, Cyclin D1. Moreover, the cells expressed IgG-lambda or IgA-lambda. We diagnosed the patient's disease as lymphoplasmacytic lymphoma (LPL). Although biclonal gammopathy with IgM elevation has been reported in LPL patients, this is the first case of IgG+IgA type biclonal gammopathy.

A 71-year-old male consulted our hospital complaining of right tonsil enlargement. Tonsillectomy was performed. Histological analysis demonstrated AL amyloid deposits obliterating the tonsillar structures. Foreign body giant cell reaction and morphologically normal plasma cells mainly containing light chain lambda were present around the amyloid. Serum immunoglobulin levels were normal and M-proteins were not detected. Urinary test for Bence Jones proteins was negative. Bone marrow aspirates did not demonstrate any signs of multiple myeloma. Amyloidosis of the stomach, colon, heart, and skin was ruled out. We report an extremely rare case of amyloidosis localized in the tonsil and review the literature.