Anaplastic Lymphoma Kinase (ALK) and p53 Are Potentially Useful Markers to Distinguish Inflammatory Myofibroblastic Tumor

Aims: Inflammatory myofibroblastic tumor (IMT) of the urinary bladder is a clinically and histologically uncommon benign tumor that can be easily mistaken for a malignant neoplasm. We sought to determine whether immunohistochemical staining would be evaluated IMT of the urinary bladder. We have also shown the literatures that imminohistochemical staining of IMT was investigated to distinguish malignant lesions using PubMed data base. Methods: Immunohistochemical staining, including anaplastic lymphoma kinase (ALK), p53, cytokeratin, vimentin, desmin, alpha-smooth muscle actin, myoglobin, smooth muscle myosin and S100, was carried out on serial sections from archival specimens of three patients who underwent transurethral resection and partial cystectomy. Results: Immunohistchemical staining in all patients was positive for ALK and weak positive for p53 protein. In the literatures, positive rates of ALK and p53 inthe IMT of the urinary bladder were 60.9% and 53.1%, respectively. Sarcoma and carcinosarcoma were shown in the pathological specimens with negative ALK and strongly positive p53 inthe same data base. Conclusions: Both ALK and p53 were potentially useful protein markers to distinguish between IMT and sarcoma. However, this study was small sample size. Further study was warranted an investigation of the availability of these proteins in IMT.

Myristoylated Alanine-Rich C Kinase Substrate Accelerates TNF-<i>α</i>-Induced Apoptosis in SH-SY5Y Cells in a Caspases-6 and/or -7-Dependent Manner

Cell proliferation, differentiation, and the elimination of unnecessary cells by apoptosis occur in the development of the nervous system. It is reported that brain dysplasia appears as the results of myristoylated alanine-rich C kinase substrate (MARCKS) knockout or the mutant mouse. We therefore expect that MARCKS participates in the development of the nervous system. However, the mechanism underlying such participation has not been identified. In this study, we observed the effects of the overexpression of MARCKS or unphosphorylatable MARCKS on cell proliferation and TNF-α-induced apoptosis in neuroblastoma SH-SY5Y cells. Furthermore, we restrained MARCKS expression by the RNAi method. In the results, MARCKS-overexpressing cells and not unphosphorylatable MARCKS-overexpressing cells showed increased cell proliferation rates. On the other hand, the RNAi decreased the proliferation of MARCKS-knocked down SH-SY5Y cells. These results indicated that MARCKS-overexpressing cells were more sensitive to TNF-α than normal SH-SY5Y cells. Moreover, in MARCKS-overexpressing cells TNF-α-induced apoptosis was inhibited by caspase-6 and -7 inhibitors but not by caspase-3 inhibitor. These results suggested that MARCKS participated in TNF-α-induced apoptosis in a caspase-6 and/or -7-dependent manner.

小柴胡汤腹腔注射对小鼠腹腔巨噬细胞的激活

作者发现小柴胡汤作为一个多克隆B细胞活化剂而促进B细胞分裂,对SRBC的抗体反应呈免疫佐剂效果。最近,川喜田等的工作表明小柴胡汤腹腔注射能引起腹腔和脾未成熟的IgM+、IgD-iB细胞明显积聚。然而,小柴胡汤激活巨噬细胞的免疫药理作用还不清楚。本文作者研究了小柴胡汤在体内对小鼠巨噬细胞的活化效果。 实验用9～10周龄的C3H/He鼠、BALB/C×DBA/2的F1（COF1）鼠或20周龄C3H/HeJ鼠,均为雌性。CDF1鼠的实验在无特殊病原体环境进行。

日本汉方药中对免疫和神经系统的活性成分和可能作用机制(英文)

Abatract:Many of the diseases are multifactorial and still difficult to treat by only modern medicines,however Kampo medicines,which contain multiple ingredients due to component herbs,seem to be suitable for recovery of such complicated symptoms because these ingredients may attack multiple target sites to improve the symptoms caused by disturbance of whole body system.In Japan,148 standardized Kampo pharmaceutical preparations have been developed,and covered by national health insurance system,and more than 80% practicing physicians have experience of using the Kampo medicines for the treatment depending on patient′s clinical situation,either separately or to complement modern western medicines.In present paper,I′d like to introduce few examples of our studies on elucidations of pharmacological actions and active ingredients of Kampo medicines as multi-ingredients drugs.Although Kampo medicines have been taken orally as the decoction,their hot water extracts contain not only low molecular ingredients but also high molecular ingredients like polysaccharides.Our recent studies suggest that Hochuekkito(HET:Bu-Zhong-Yi-Qi-Tang)prevents respiratory infection through potentiation of mucosal immune system in upper respiratory tract,and certain pectic polysaccharides were identified to involve in potentiation of the immune response.It was also clarified that synergic action of two stereoisomers are required for the oral adjuvant activity of Shoseiryuto(SST:Xiao-Qing-Long-Tang).Like examples of the studies of Kososan(KS:Xiao-Su-San)and SST,proteomic analysis of disease model animals using agarose 2-DE is also useful another approach to find target protein candidates.These accumulated comprehensive results may contribute for more evidence-based clinical application.

Comparison of the Normal, Preperimetric Glaucoma, and Glaucomatous Eyes with Upper-Hemifield Defects Using SD-OCT

Purpose: We compared the thickness of circumpapillary retinal nerve fiver layer (cpRNFL) and macular ganglion cell layer with inner plexiform layer (GCL + IPL) using Cirrus HD-OCT (Ver.6.0: Carl Zeiss). Materials and Methods: This study included 12 eyes of normal controls, 10 eyes of preperimetric glaucoma (PPG) with loss of RNFL either in superior or in inferior hemisphere without visual field defects, and 22 eyes of glaucoma eyes with visual field defects restricted to upper hemifield (UHFD: early 10 eyes, severe 12 eyes). The cpRNFL thickness analyzed from disk center by dividing into 12 sectors. The GCL + IPL thickness analyzed from central fovea by dividing into six sectors. Both compared between normal eye group and other 3 groups using the average value of each sectors. Result: The cpRNFL and the GCL + IPL thickness were obviously thin as compared with normal eyes. Conclusion: Even if it is in the state where abnormalities are not detected using the Humphrey field Analyzer, it is suggested that the early structural change of glaucoma has already arisen.

Dephosphorylation of myristoylated alanine-rich C kinase substrate accelerates wound-induced migration of SH-SY5Y cells

Inflammation, which is induced after infection of bacteria and tissue injury, is one of the important early stages of wound healing. Bradykinin is increased during acute and chronic inflammation. We previously reported that bradykinin stimulation induces dephosphorylation of myristoylated alanine-rich C kinase substrate (MARCKS) after phosphorylation by ROCK leading neurite outgrowth in neuroblastoma SH-SY5Y cells. In this report we showed that knock-down of MARCKS by RNAi reduced cell migration. Wild-type MARCKS-overexpressed SH-SY5Y cells migrated faster than the control cells. Unphosphorylatable MARCKS-overexpressed cells notably migrated fast. Moreover, chronic MARCKS dephosphorylation by a ROCK inhibitor HA-1077 promoted the cell migration, on the other hand a PKC inhibitor Ro-31-8220 did not. After wounding, MARCKS was transiently phosphorylated and dephospho-rylated by 20 min. Immunocytochemistry showed that the dephosphorylated MARCKS was localized at neurite tips. These findings suggest that MARCKS dephosphorylation is important in wound-induced migration of SH-SY5Y cells. It indicates the possibility that MARCKS is associated with wound repair after inflammation.

Developing a Scale for Measuring Uncertainty in Patients with Unruptured Intracranial Aneurysms Undergoing Endovascular Coiling

A measure called the Uncertainty in Unruptured Intracranial Aneurysm Patients Undergoing Endovascular Coiling Scale (UUIACS) was developed and its validity and reliability were examined. The 49 questions that comprised the original draft of the UUIACS were created based on interview data. Based on data from 172 participants, exploratory and confirmatory factor analyses were conducted. As a result of exploratory factor analysis, the UUIACS e retained 17 items and extracted four factors (“Lack of decision-making cues”, “Lack of information and complexity of information interpretation”, “The ambiguous nature of the disease”, and “The unpredictable living with UIA”). All of the UUIACS items showed adequate internal consistency. Between the UUIAC scale and the Universal Uncertainty in Illness Scale (UUIS), the Health Locus of Control (HLC) scale, and the SF-36v2&reg (Japanese version), positive correlations were found between the UUIACS and UUIS, and the HLC scale at a 1% significance level indicating concurrent validity. According to confirmatory factor analysis, the UUIACS had an acceptable goodness of fit. Given these findings, the UUIACS was judged to have satisfied the criteria for use in a clinical setting, although further investigation was required.

Comparison of glycemic variability between basal-bolus and premixed insulin therapy

Background: Several studies have shown that twice-daily injections of premixed insulin analogs (MIX) could achieve comparable HbA1c levels to basal-bolus (BB) therapy. However, HbA1c does not necessarily reflect short-term glycemic fluctuations that may contribute to the onset or progression of diabetic complications. Therefore, in this study, we compared MIX and BB therapies in terms of their effects on glycemic variability. Methods: We performed a crosssectional observational study of patients attending our outpatient clinics to compare the effects of two insulin regimens on glycemic variability. We recruited patients treated with MIX or BB with HbA1c < 8.4%. A total of 27 patients (11 treated with BB and 16 treated with MIX) were enrolled and wore a continuous glucose monitor (CGM) for 72 h, while continuing their usual lifestyle and insulin doses. Results: No significant differences in CGM-determined glycemic markers were observed between the two groups. However, the post-lunch duration of glucose levels > 180 mg/dL (t > 180) was significantly shorter with BB therapy (88 ± 76 min) than with MIX therapy (145 ± 54 min;p < 0.05). After classification according to HbA1c levels, markers of glycemic variability were better in patients treated with BB than in those treated with MIX in better control group. Conclusion: These results suggest that BB therapy achieves better glucose profiles than MIX therapy in patients with type 2 diabetes, particularly after lunch.

Rodent Macrophage Select Vin Blank Together with Vin Rouge According to SO Level <i>in Situ</i>

Phagocytic cells are known as multi potent activities for its ontogenical and phylogenetical aspect. One of the strike actions is super oxide anion known as critical role of the cell when microorganisms invade inside the cytoplasm. This agent sometimes triggered as a serious factor in the site of inflammation. There are many papers which concerned newly accessed anti-oxidative substances. However, many reports only focused on the molecular basis in vitro, suggesting vin rouge for their rich content of polyphonol rather than vin blank with a French paradox. We had been suggested that the key factor for oxidative stress needs to be discussed first for immunological standpoint, such as in phagocytic cell in situ. With this assay system in rodent macrophage, SO level was about the same for both groups administered vin blank as well as vin rouge.

Overall Estimation of Anti-Oxidant Activity by Mammal Macrophage

Super oxide anion had been known as a critical role in a phagocytic cell when microorganisms invade inside. However, this radical seldom need for a clean environment such in a developed country. Moreover, this agent sometimes triggered as serious factor in the site of inflammation. There are many papers which concerned newly found anti-oxidative substances. However, many reports only focused on the molecular basis in vitro, how oxidative stress damaged the human tissue. The key factor for oxidative stress needs to be discussed first for immunological standpoint, such as armed reduction treaty. In other words, minimum armed force is necessary for an invasion by outer or inner irregular cells. On the basis of phagocytic function of the cell in vivo, we tried to access anti-oxidative activity by employing peritoneal murine macrophage and human monocyte. So, the purpose of this study is to establish a conventional but tailor made system for which is useful for evaluating the anti-oxidative activity to different constition. Especially in human peripheral monocyte, the sample was tested each constitution such in granulocyte rich type and lymphocyte rich typ. In order to establish the direct estimation by phagocytic cell, a murine peritoneal exudates cell suspension was obtained after one week of oral administration with anti-oxidative sample. The samples we selected were traditional Chinese medicine, Shi-Quan-Da-Bu-Tang (SDT), Bu-Zong-Ye-Qi-Tang (BYT) and Shao-Chin-Rong-Tang sCRT), Agaricus, Chlorella, and Propolis. The amount of generated super oxide anion was calculated through measuring the optical density change of this supernatant using cytochrome-C which was reduced by generated super oxide anion in the phagocytic cell. The generated super oxide anion after administration of Agaricus, Chlorella, SDT, BYT, sCRT and Propolis were 2.64, 1.95, 1.24, 1.25, 2.88, 2.55 × 10﹣5 mmol/ml, respectively, whereas 2.85 × 10﹣5 mmol/ml in control group. The fermented sample from SDT, BYT, sCRT and microfied Propolis were also tested and the value of super oxide anion were 0.62, 0.84, 1.50 and 2.55 × 10﹣5 mmol/ml, respectively. In this report, we discussed the direct method for measuring the generated super oxide anion of macrophages, especially from different constitution.

Regulative effect for natural killer cell by hot spring hydrotherapy—Quantitative and qualitative discussion

Along with maintaining immune competent cells, one of the purposes of cancer patient is regulating the first line of defense for survival. Moreover, the factors that influence the acquired immune activity are systemic metabolic disorder in diabetes, malnutrition, extreme exhaustion, stresses, aging and medical side effect such as chemotherapy. So we have to select appropriate menu to regulate immune function through leukocyte storage. Especially, NK cell is first line of defense against virus infected cell and/or tumor cell. In order to recommend the effect of hot spring hydrotherapy on the peripheral white blood cells as a parameter for complementary and alternative medicine (CAM) within a short period, the granulocytes, lymphocytes, lymphocyte subsets in their peripheral blood were monitored. In the young group (35 or less), the number of granulocytes clearly decreased after hydrotherapy. In the older group (over 35 years old), the total number of leukocytes and lymphocytes significantly increased after the therapy. Using a FACScan, we investigated CD2, CD4, CD8, CD16, CD19 and CD56 positive cell as well as the cytokine containing cell, as direct assessment for immunological preparation (IFN-γ, IL-4 and IL-1β). After hydrotherapy, the CD16+ cells also increased outstandingly. However, the CD19+ cells clearly decreased. Moreover, there was a decrease in the cytokine-producing cell count of subjects whose blood cells had a higher number of the cell level before. All of the results indicated that hot spring hydrotherapy could regulate the immune system even in normal human. Like this, blood hormonal level was measured in order to know the mechanism of the quantitative and qualitative, adjustment of the leucocytes becomes possible within a short term. As a result of measuring, 7 kinds of hormone including mainly adrenocortical hormones, adrenalin level reduced in constitution dependent manner. From these results, it was suggested that the quantitative and qualitative fluctuation of the leukocyte by the hydrotherapy is closely related to dynamics of blood hormonal level.

Clinical Effects of a Plant Extract Mixture Containing Rhus verniciflua and Other Herbs in Tumor Bearing Dogs

Dog owners are increasingly seeking treatment when their pets develop cancers. As in human cancer patients, dogs with cancer are commonly treated with complementary and alternative therapies, including herbal medicines and nutritional supplements. A novel antitumor agent was developed from six different herbs including Rhus verniciflua (Rv-PEM01). The components were established from traditional herbal medicine and designed to affect antitumor activity and maintain host immune function. Previous studies identified anti-proliferative activity in human, murine and canine cancer cell lines. In this clinical study the safety and tolerability of Rv-PEM01 were evaluated in pet dogs with spontaneously occurring cancers. Twelve dogs were treated orally daily for 30 days in escalating dose (4 - 10 mg/kg orally once daily) cohorts. Rv-PEM01 was well tolerated;only transient mild elevations in BUN were observed in 2 dogs. Although tumor response was not a primary endpoint for this study, stable disease was maintained for 30 days in 5 (42%) of the dogs. In conclusion, Rv-PEM01 was found to be safe and well tolerated in the dosage range tested. Future studies should evaluate higher dosages of Rv-PEM01 in dogs with cancer, and specifically address other potential benefits of Rv-PEM01 in canine cancer patients, including correlative assessments of immune function, quality of life and owner satisfaction.

Antitumor Effects and Acute Oral Toxicity Studies of a Plant Extract Mixture Containing <i>Rhus verniciflua</i>and Some Other Herbs

A novel antitumor agent was developed from six kinds of herbs containing Rhus verniciflua (Rv-PEM01). The components were traditionally established for each formula for traditional medicine. The formula was designed to affect antitumor effect as well as maintain host immune functions. First, we investigated the antiproliferative activities of Rv-PEM01 on human and canine tumor cell lines in vitro, and on antitumor effects using BALB/cAJcl-nu/nu mice in vivo. Acute oral toxicity of Rv-PEM01 was also investigated in vivo in ddY mice. Rv-PEM01 exhibited antiproliferative activities against PC-3 (IC50: 0.328 ± 0.081 mg/ml), A549 (IC50: 0.520 ± 0.070 mg/ml), D-17 (IC50: 0.124 ± 0.037 mg/ml) and MRC-5 (IC50: 0.505 ± 0.058 mg/ml) cells. Luteolin 7-β-D-glucopyranoside and apigenin 7-β-D-glucopyranoside were identified as the main active compounds in Rv-PEM01 by HPLC analysis. The single dose toxicity study of Rv-PEM01 did not result in any deaths or abnor-malities in daily behavior, body weight gain, or anatomical observations at necropsy. Thus, so we could not calculate the 50% lethal dose (LD50) in mice, but it would be higher than 5.0 g/kg. Treat- ment with Rv-PEM01 at a dose of 2.5 g/kg tended to show antitumor activities on mice bearing Colon26 tumors compared with the control group. It was concluded that the formula was a safe antitumor agent with no side effects on mouse physiological function as judged by survival and organ weight.

结核感染若干年后特异性γ-干扰素应答减弱

假设:随着时间的流逝,结核感染者记忆性T细胞对特异性抗原的应答减弱。背景:很多证据表明特异性全血γ-干扰素(IFN-γ)检测试验Quanti-FERON-TB Gold (QFT-G)能够检测出近期结核感染者,并具有较高的灵敏度和特异性。目的:我们应用此项技术检测日本一个社区的成人群体以确定其流行病学应用价值。方法:1559例参加定期体检的健康志愿者入选本研究。结果:40～49岁人群QFT-G的阳性率为3.1%,50～59岁人群的阳性率为5.9%,60～69岁人群的阳性率为9.8%。作者根据一系列研究估计40～49岁、50～59岁、60～69岁的预期结核感染率分别为11.1%、29.6%、53.1%。预期阳性率与实际阳性率之间存在的巨大差异提示结核感染后随着时间的延长IFN-γ应答减弱。那些经胸部X线证实存在陈旧性结核病灶者阳性率大大低于100%。结论:特异性IFN-γ应答可能在结核感染后随着时间的推移大大地减弱。需要在感染者中长期地纵向研究细胞介导的免疫动力学。

Recent evidence for subcutaneous drains to prevent surgical site infections after abdominal surgery:A systematic review and metaanalysis

BACKGROUND Surgical site infections(SSIs)increase mortality,hospital stays,additional medical treatment,and medical costs.Subcutaneous drains prevent SSIs in gynecological and breast surgeries;however,their clinical impact in abdominal surgery remains unclear.AIM To investigate whether subcutaneous drains were beneficial in abdominal surgery using a systematic review and meta-analysis.METHODS The database search used PubMed,MEDLINE,and the Cochrane Library.The following inclusion criteria were set for the systematic review:(1)Randomized controlled trial studies comparing SSIs after abdominal surgery with or without subcutaneous drains;and(2)Studies that described clinical outcomes,such as SSIs,seroma formation,the length of hospital stays,and mortality.RESULTS Eight studies were included in this meta-analysis.The rate of total SSIs was significantly lower in the drained group(54/771,7.0%)than in the control group(89/759,11.7%),particularly in gastrointestinal surgery.Furthermore,the rate of superficial SSIs was slightly lower in the drained group(31/517,6.0%)than in the control group(49/521,9.4%).No significant differences were observed in seroma formation between the groups.Hospital stays were shorter in the drained group than in the control group.CONCLUSION Subcutaneous drains after abdominal surgery prevented SSIs and reduced hospital stays but did not significantly affect seroma formation.The timing of drain removal needs to be reconsidered in future studies.

Inhibition of tyrosinase activity and melanine pigmentation by 2-hydroxytyrosol

2-Hydroxytyrosol(2-HT),originally reported as a synthetic compound,was isolated for the first time as a fungal metabolite.2-HT was found to inhibit mushroom tyrosinase with an IC_(50) value of 13.0 mmol/L.Furthermore,2-HT dose-dependently inhibited tyrosinase activity(IC_(50),32.5 mmol/L)in the cell-free extract of B16 melanoma cells andα-melanocyte stimulating hormone(α-MSH)-stimulated melanin formation in intact B16 melanoma cells.

New cyslabdans B and C, potentiators of imipenem activity against methicillin-resistant Staphylococcus aureus produced by Streptomyces sp. K04-0144

From a further purification study,two new congeners designated cyslabdans B(1)and C(2)were isolated along with previously reported cyslabdan(cyslabdan A(3)in this study)from the culture broth of Streptomyces sp.K04-0144.The structure was elucidated by various spectroscopy including NMR,revealing that 1 and 2 was 18-hydroxy and 10-methoxy cyslabdan,respectively.Compounds 1 and 2 were found to potentiate imipenem activity against methicillinresistant Staphylococcus aureus by 123 fold and 533 fold,respectively.Comparison with the activity of compound 3 indicated that the introduction of a hydrophilic group at the dimethyl moiety of the decalin ring was unfavorable for its activity.

Beauvericin counteracted multi-drug resistant Candida albicans by blocking ABC transporters

Multi-drug resistance of pathogenic microorganisms is becoming a serious threat,particularly to immunocompromised populations.The high mortality of systematic fungal infections necessitates novel antifungal drugs and therapies.Unfortunately,with traditional drug discovery approaches,only echinocandins was approved by FDA as a new class of antifungals in the past two decades.Drug efflux is one of the major contributors to multi-drug resistance,the modulator of drug efflux pumps is considered as one of the keys to conquer multi-drug resistance.In this study,we combined structure-based virtual screening and whole-cell based mechanism study,identified a natural product,beauvericin(BEA)as a drug efflux pump modulator,which can reverse the multi-drug resistant phenotype of Candida albicans by specifically blocking the ATP-binding cassette(ABC)transporters;meantime,BEA alone has fungicidal activity in vitro by elevating intracellular calcium and reactive oxygen species(ROS).It was further demonstrated by histopathological study that BEA synergizes with a sub-therapeutic dose of ketoconazole(KTC)and could cure the murine model of disseminated candidiasis.Toxicity evaluation of BEA,including acute toxicity test,Ames test,and hERG(human ether-a-go-go-related gene)test promised that BEA can be harnessed for treatment of candidiasis,especially the candidiasis caused by ABC overexpressed multi-drug resistant C.albicans.

Fungal pyrrolidine-containing metabolites inhibit alkaline phosphatase activity in bone morphogenetic protein-stimulated myoblastoma cells

Fibrodysplasia ossificans progressiva(FOP)is a rare autosomal dominant congenital disorder characterized by progressive heterotopic ossification in muscle tissues.A constitutively activated mutation of a bone morphogenetic protein(BMP)receptor,ALK2,has been identified in patients with FOP.We report here that four structurally related compounds,lucilactaene,hydroxylucilactaene,NG-391 and NG-393,produced by fungal strain Fusarium sp.B88,inhibit BMP signaling in vitro.Alkaline phosphatase activity,a marker enzyme of osteoblastic differentiation,was decreased in C2C12 myoblasts stably expressing mutant ALK2 by treatment with those compounds with IC_(50) values of 5.7,6.8,6.9 and 6.1 mM,respectively.Furthermore,NG-391 and NG-393 inhibited BMP-specific luciferase reporter activity,which is directly regulated by transcription factor Smads,with IC50 values of 1.4 and 2.1 mM,respectively.These findings suggest that these fungal metabolites may provide a new direction in the development of FOP therapeutics.