Medium-term results of neoadjuvant systemic chemotherapy using irinotecan, 5-fluorouracil, and leucovorin in patients with locally advanced rectal cancer.

AIMS: The aim of this study was to evaluate the usefulness of neoadjuvant systemic chemotherapy using irinotecan, 5-FU, and leucovorin (LV) for the treatment of locally advanced rectal cancer, which was a powerful ploychemotherapy in those days in Japan. METHODS: Between 2001 and 2004, 26 patients with T3 or T4 and N0-2 non-metastatic resectable rectal cancer were selectively enrolled in this study. Neoadjuvant chemotherapy consisted of two cycles of irinotecan (80 mg/m²), 5-FU (500 mg/m²), and LV (250 mg/m²) on days 1, 8, and 15 for 4 weeks. Surgical resection was performed in all the patients 2-4 weeks after the completion of chemotherapy. RESULTS: Overall down-staging was observed in 15 patients. T level and N level down-staging were observed in 12 and 13 patients, respectively. A pathological complete response was observed in one patients. The median follow-up period was 75 months (range, 8-97 months). Recurrences occurred in 5 patients including pelvic relapses in 3 and distant metastases in 2. The 5-year relapse-free and overall survival rates were 74% and 84%, respectively. CONCLUSIONS: Neoadjuvant systemic chemotherapy comprised of a combination of multi-drugs as irinotecan, 5-FU, and LV may be beneficial to the prognoses of patients with locally advanced rectal cancer.

Administration of isoferulic acid improved the survival rate of lethal influenza virus pneumonia in mice.

BACKGROUND: Isoferulic acid (IFA) is a main active ingredient of the rhizoma of Cimicifuga beracleifolia, which is used frequently in Japanese traditional medicine as an anti-inflammatory drug. It has been revealed that IFA inhibits the production of macrophage inflammatory protein-2 (MIP-2), which is a murine counterpart of the chemokine family that may contribute to the pathogenesis of inflammatory diseases through the chemotactic activity for inflammatory and immune effector cells. AIM OF THE STUDY: In this study, we investigated the therapeutic effect of IFA on the progression of lethal influenza virus pneumonia in mice by comparison with that of dexamethasone (DX), a potent inhibitor for various inflammatory cytokines including MIP-2. METHODS: Mice were infected by intranasal inoculation of influenza virus under ether anesthesia. The IFA or DX was given by oral administration once daily for 4 days after infection. After infection, the survival rate and the change in body weight were daily monitored. RESULTS: IFA administration markedly improved the survival rate and body weight loss of influenza virus-infected mice in a suitable dose range (0.5 mg/day). However, DX administration did not show a beneficial effect at any dose. CONCLUSION: These data suggested that IFA is a novel tool not only for the intervention therapy, but also for the studies on the pathogenesis of influenza virus-induced pneumonia.

Inhibitory effect of (+)-catechin on the growth of influenza A/PR/8 virus in MDCK cells.

We investigated whether (+)-catechin, a building block of tannins contained in the extract of Ephedrae herba (EHext), exerts an inhibitory effect on the acidification of intracellular compartments such as endosomes and lysosomes (referred to as ELS), and thereby inhibits the growth of influenza A PR/8/34 (PR8) virus (H1N1 subtype) in Madin-Darby canine kidney cells. The vital fluorescence microscopic study with acridine orange showed that 1-h treatment with (+)-catechin inhibited the acidification of ELS in a concentration-dependent manner (1.0-10.0 mM). Moreover, the growth of PR8 virus was inhibited markedly when the cells were treated with (+)-catechin (1.25-10.0 mM) for 1 h immediately after infection, or treated within as little as 5 to 10 min after infection. Conversely, virus growth resumed within 3 h concomitantly with the reappearance of acidified ELS after removal of (+)-catechin. Similar to EHext, (+)-catechin inhibited both the acidification of ELS and the influenza virus growth. It suggests that (+)-catechin is one of the active components in EHext.

Nitric oxide-mediated antitumor activity induced by the extract from Grifola frondosa (Maitake mushroom) in a macrophage cell line, RAW264.7.

We have investigated D-fraction (MDF) extracted from Grifola frondosa (Maitake mushroom) on the inducible nitric oxide synthase (iNOS)-mediated nitric oxide (NO) production in RAW264.7 (RAW) cells, a murine monocyte/macrophage cell line, with special reference to antitumor activity of MDF against human hepatoma-derived huH-1 cells. MDF could induce iNOS mRNA expression in RAW cells in a dose range of more than 30 microg/ml, but the effect of 10 microg/ml of MDF was negligible. The iNOS mRNA expression induced by 100 microg/ml of MDF was 6 hrs later, but lasted for a longer time than that of lipopolysaccharide (LPS), a representative iNOS inducer. Although iNOS mRNA levels in MDF-stimulated cells were almost equal to LPS-stimulated cells at the peak time, the cumulative amount of nitrite was only about 50% compared with that of LPS-treated cells. When huH-I cells were cultured in MDF containing media in a 24-well plate with inserted porous bottom in the presence or absence of RAW cells, the viability of huH-1 cells decreased significantly only in the presence of RAW cells in MDF dose-dependent manner. This antitumor activity of RAW cells in the presence of MDF was abolished or attenuated by the addition of L-NAME, a NOS inhibitor, confirming that this phenomenon is due to iNOS-mediated NO production by RAW cells, but not direct cytotoxic activity of MDF against huH-1 cells. These data suggest that MDF is a novel inducer for iNOS which contributes at least in part to antitumor activity of MDF.

Deformation of the ventrolateral medulla oblongata by subarachnoid hemorrhage from ruptured vertebral artery aneurysms causes neurogenic pulmonary edema.

The occurrence of neurogenic pulmonary edema (NPE) associated with subarachnoid hemorrhage (SAH) due to ruptured aneurysm was analyzed in 48 consecutive patients. Correlations of the location of the aneurysm, clinical grade, amount of subarachnoid clot, and severity of NPE were examined. NPE was observed in 29.4% of all SAH cases, but the incidence was significantly higher in cases of ruptured vertebral artery (VA) aneurysm. Clinical grade, severity of NPE, and deformation of the medulla oblongata were studied in the five cases of ruptured VA aneurysm. Deformation of the ventrolateral medulla oblongata was observed in all patients. Asymmetry index of the medulla oblongata measured on the axial computed tomography scan was correlated with the severity of NPE. Severity of NPE tended to correlate with deformation of the medulla oblongata. NPE associated with ruptured VA aneurysm is caused by deformation of the ventrolateral site of the medulla oblongata by the localized hemorrhage.

Enhanced expression of inducible nitric oxide synthase by Juzen-taiho-to in LPS-activated RAW264.7 cells, a murine macrophage cell line.

We have investigated the effect of Juzen-taiho-to (TJ-48) on inducible NO synthase (iNOS) expression and nitric oxide (NO) production in RAW264.7 cells, a murine macrophage cell line. TJ-48-lipopolysaccharide (LPS) combination induced iNOS mRNA expression earlier, stronger and remained longer that paralleled but with a higher NO production compared to LPS stimulation. TJ-48 itself showed no inducible effect either on NO production or iNOS mRNA expression. This phenomenon could be considered to contribute, at least in part, to the beneficial effects of TJ-48 through the iNOS-mediated activation of biodefense mechanism.

Inhibitory effect of ferulic acid and isoferulic acid on the production of macrophage inflammatory protein-2 in response to respiratory syncytial virus infection in RAW264.7 cells.

We investigated the effect of ferulic acid (FA) and isoferulic acid (IFA), which are the main active components of the rhizoma of Cimicifuga heracleifolia (CH), an anti-inflammatory drug used frequently in Japanese traditional medicine, on the production of macrophage inflammatory protein-2 (MIR-2) in a murine macrophage cell line, RAW264.7, in response to respiratory syncytial virus (RSV) infection. Following the exposure of cells to RSV for 20h, the MIP-2 level in condition medium was increased to about 20 ng/ml, although this level in mock-infected cells was negligible. In the presence of either FA or IFA, RSV-infected cells reduced MIP-2 production in a dose-dependent manner. These data suggest that FA and IFA might be responsible, at least in part, for the anti-inflammatory drug effect of CH extract through the inhibition of MIP-2 production.

Inhibitory effect of Ephedrae herba, an oriental traditional medicine, on the growth of influenza A/PR/8 virus in MDCK cells.

Using several herbal extracts, we investigated whether certain Kampo medicines exert an inhibitory effect on the acidification of intracellular compartments such as endosomes and lysosomes (referred to as ELS), and thereby inhibit the growth of influenza A virus in Madin-Darby canine kidney cells. The vital fluorescence microscopic study showed that the extract of Ephedrae herba (EHext) among five herbal extracts inhibited acidification of endosomes and lysosomes in a concentration-dependent manner (100-400 microg/ml). Moreover the growth of influenza A/PR/8/34 (H1N1) (PR8) virus was inhibited when the cells were treated with EHext for 1 h immediately after infection, or treated as early as 5-10 min after infection. Conversely, virus growth resumed concomitantly with the reappearance of acidified ELS after removal of EHext. The fact that the inhibitory effect of EHext was completely or partially reversed by FeCl3, a tannin-reactive agent, strongly suggests that tannin is one of the active components in the extract.

Fos protein induction in the medullary dorsal horn and first segment of the spinal cord by tooth-pulp stimulation in cats.

Electrophysiological studies using the single neuron recording technique have led to the hypothesis that nociceptive neurons in the medullary dorsal horn (MDH) and the first segment of the spinal cord (C1) encode the stimulus intensity of noxious stimuli applied to the tooth pulp. The present study utilized the Fos protein technique in combination with electrical and chemical stimulation of the tooth pulp to test this hypothesis. Upper canine tooth-pulp stimulation with intensities just above the threshold stimulus intensity for evoking the jaw-opening reflex (JOR) did not produce a clear expression of Fos protein-like immunoreactive (LI) cells in the MDH and C1 of cats. Fos protein-LI cells were mainly found in the superficial laminae (laminae I-II) of the MDH and C1 after tooth-pulp stimulation of 200% of the JOR threshold intensity. When higher intensities (400-600% of the JOR threshold intensity) or mustard oil were applied, Fos protein-LI cells were also found in laminae III-IV as well as in laminae I-II. The number of Fos protein-LI cells significantly increased when 600% of the JOR threshold intensity or mustard oil was applied. Furthermore, the rostro-caudal distribution of Fos protein-LI cells was greater following increases in stimulus intensities and the greatest after mustard oil application. These data suggest that the change in number and spatial arrangement of nociceptive neurons in the MDH and C1 reflect changes in the encoding of the stimulus intensity applied to the tooth pulp.

Inhibitory effect of ferulic acid and isoferulic acid on murine interleukin-8 production in response to influenza virus infections in vitro and in vivo.

We investigated the effect of ferulic acid (FA) and isoferulic acid (IFA), which are active components of the rhizoma of Cimicifuga species used frequently as anti-inflammatory drugs in Japanese Oriental medicines, on murine interleukin-8 (IL-8) production in response to influenza virus infections in vitro and in vivo by antibody-sandwich enzyme-linked immunosorbent assay. In the in vitro study, the murine macrophage cell line RAW 264.7 was infected with influenza virus at a dose of 10 plaque forming units (PFU)/cell and cultured in the presence or absence of drugs. Both FA and IFA reduced the IL-8 levels in the 20-h conditioned medium in comparison with control in a dose-dependent manner. The effect of IFA was greater than that of FA: IL-8 levels were reduced to 43% and 56% of the control in the presence of 100 micrograms/ml of IFA and FA, respectively. In the in vivo study, mice were infected with 1,000 PFU of virus and received daily oral administrations of Cimicifuga heracleifolia extract (5 mg/mouse/day), FA (0.5 mg/mouse/day), IFA (0.125 mg/mouse/day), or phosphate buffered saline. The three drugs showed a tendency to reduce IL-8 levels in bronchoalveolar lavage (BAL) obtained 2 days after infection. Moreover, both FA and IFA also significantly reduced the number of exuded neutrophils into BAL. However, the drug administrations did not affect the virus yields in BAL. These data suggest that FA and IFA are novel and potent inhibitors of murine IL-8 production and might act as one of the main components of anti-inflammatory rhizoma of Cimicifuga species.

Antiviral traditional medicines against herpes simplex virus (HSV-1), poliovirus, and measles virus in vitro and their therapeutic efficacies for HSV-1 infection in mice.

One hundred forty-two kinds of traditional medicines, which have been historically used in China, Indonesia, and Japan, were examined for the antiviral activity of their hot water (HW) extracts against herpes simplex virus type 1 (HSV-1), poliovirus type 1, and measles virus by plaque reduction assay. Thirty-two, 55, and 30 HW-extracts of them showed anti-HSV-1, antipoliovirus, and anti-measles virus activities, respectively. Among the 32 HW-extracts with anti-HSV-1 activity, 3 HW-extracts had anti-HSV-1 activity alone and the others showed anti-HSV-1 activity with anti-poliovirus and/or anti-measles virus activities. The 32 HW-extracts were further examined for their therapeutic efficacies of HSV-1 infection in mice. The mice were infected cutaneously with HSV-1 and HW-extracts were orally administered three times daily. Twelve HW-extracts, currently used for the treatment of various diseases other than viral infection, were found to be significantly effective in limiting the development of skin lesions and/or in prolonging the mean survival times of HSV-1-infected mice. These results suggested that 12 of 142 HW-extracts that exhibited therapeutic efficacy in an animal infection model were possible candidates for anti-HSV-1 traditional medicine.

[Solubilizing effect and inclusion reaction of cyclic bisdesmosides from tubers of Bolbostemma paniculatum].

Tubeimosides I, II and III (cyclic bisdesmosides) have been isolated from Chinese cucurbitaceous crude drug Tu-bei-mu, a tuber of Bolbostemma paniculatum (Maxim.) Franquet. Solubilizing effects of these cyclic bisdesmosides on water insoluble or less-soluble compounds were examined. It was revealed that cyclic bisdesmosides were effective on increasing the solubility of Yellow OB, dl-alpha-tocopherol and saponin A from Sapindus mukurossi. The critical micell concentration (cmc) and association number as well as diameter of micell of tubeimoside I in water were also measured. The interaction of tubeimoside I with 1-anilino-8-naphthalene-sulfonate (ANS) in aqueous solution was investigated photometrically. It was observed that tubeimoside I strongly enhanced the intensity of fluorescence of ANS, suggesting the significant formation of inclusion complex.

Cytotoxic and antiherpetic activity of phloroglucinol derivatives from Mallotus japonicus (Euphorbiaceae).

The phloroglucinol derivatives isolated from Mallotus japonicus Muell. Arg. (Euphorbiaceae) and their derivatives were evaluated for their capacity to produce cytotoxicity in HeLa cells and to inhibit the replication of herpes simplex virus type 1 (HSV-1). The characterizations of an isolated new acetophenone, mallophenone (16), and cyclization products of mallotojaponin (1), isomallotochromene (17), mallotochroman (18) and isomallotochroman (19), were also described. All tested derivatives inhibited the replication of HSV-1 with ED50 in the range of 88 ng--48 micrograms/ml. The derivatives 12 and 19 were found in vitro therapeutic index with 10.9 and 9.1, respectively, and they were considered to be active antivirals.

Cytological evidence for different types of cerebrospinal fluid-contacting subependymal cells in the preoptic and infundibular recesses of the frog.

Blue-green fluorescent subependymal cells with intraventricular processes were shown by the fluorescent histochemical method to be distributed from the preoptic recess to the infundibular recess of the frog hypothalamus. Electron microscopy revealed at least two types of CSF-contacting subependymal cells, type 1 containing large dense granules (about 100-200 nm in diameter) and type 2 containing small dense core vesicles (about 60-100 nm in diameter). Subsequent to fixation in permanganate solution, the small dense core vesicles in type 2 cells reacted with the fixative and consistently showed a dense content. However, the large granules in type 1 cells were mostly pale or less dense after this fixation. Two hours after intraventricular injection of 3H-dopamine, a large number of silver grains appeared only in the cytoplasm of intraventricular processes possessing dense core vesicles (type 2 cells). A few grains were also found in the perikarya. It is concluded that type 2 cells are catecholamine-storing cells. It is suggested that type 1 cells in the infundibular recess are peptidergic neurons which may secrete some hypothalamic regulating hormones of the anterior pituitary. Most of these cells in the preoptic recess belong to the neurosecretory cells of the preoptic nucleus, while some cells probably function similarly to those in the infundibular recess.

Fine-structural and functional aspects of the cerebrospinal fluid-contacting subependymal cells in the anuran hypothalamus.

Cytological characteristics of the cerebrospinal fluid (CSF)-contacting subependymal cells in the frog hypothalamus were investigated to decide whether they should be classified as neurons, sensory cells or secretory (endocrine) cells. The CSF-contacting subependymal cells exhibited morphological characteristics similar to neurons or sensory cells, which are usually supplied with axo-somatic and -dendoritic synapses from other neurons and possess somato-dendritic synapses to other neurons. Furthermore, they were provided with so-called sensory cilia. The CSF-contacting subependymal cells also exhibited several cytological secretory cell characteristics: the presence of numerous secretory granules, well-developed granular endoplasmic reticulum and Golgi apparatus in the perikaryon as well as intraventricular processes, and some figures which may be regarded as microapocrine or diacrine secretions of the secretory granules. Furthermore, the CSF-contacting subependymal cells probably sent basal cell processes to the area around the blood vessels in the neurohypophysis and, perhaps, in the infundibulum. It seemed likely that CSF-contacting subependymal cells had two or three functions of a paraneuronic nature. They were sensory to chemical or physical stimuli frmones just as endocrine cells do.