Loss of SOCS3 in the liver promotes fibrosis by enhancing STAT3-mediated TGF-beta1 production.

Recently, DNA methylation and reduced expression of the suppressor of the cytokine signaling-3 (SOCS3) gene in human hepatocellular carcinoma (HCC) patients have been reported. However, the roles of SOCS3 in HCC development in vivo have not been clarified. Using RT-PCR analysis and Western blotting, we confirmed that SOCS3 expression was reduced in HCC patients. However, reduced expression of SOCS3 occurred not only in HCC but also in nontumor regions, and this reduction was stronger as the fibrosis grade increased. Furthermore, SOCS3 levels were inversely correlated with signal transducers and activators of transcription-3 (STAT3) activation as well as transforming growth factor (TGF)-beta1 levels in the non-HCC region. To define the molecular consequences of SOCS3 silencing/STAT3 hyperactivation and liver fibrosis, we examined liver-specific SOCS3-deficient mice. We demonstrated that SOCS3 deletion in the liver resulted in hyperactivation of STAT3 and promoted ConA- and chemical-induced liver fibrosis. The expression of TGF-beta1, a mediator of fibrosis, was enhanced by SOCS3 gene deletion, but suppressed by the overexpression of a dominant-negative STAT3 or SOCS3 both in vivo and in vitro. These data suggest that TGF-beta1 is a target gene of STAT3 and could be one of the mechanisms for enhanced fibrosis in SOCS3-deficient mice. Thus, our present study provides a novel role of SOCS3 and STAT3 in HCC development: in addition to the previously characterized oncogenic potentials, STAT3 enhances hepatic fibrosis through the upregulation of TGF-beta1 expression, and SOCS3 prevents this process.

Two cases of unilateral pigmented purpuric dermatosis.

Two cases of pigmented purpuric dermatosis showing unusual distribution are reported. Case 1: 35-year-old female. A group of three asymptomatic purpuric macules developed on the ulnar side of the elbow flexure of the left forear 3-6 months before presentation. Histopathological analysis revealed a band-like inflammatory infiltrate in the Uppermost dermis and extravasation of red blood cells just beneath the epidermis, but there was no evidence of lichefaction degeneration in the basal layer of the epidermis. Case 2: 26-year-old female. Asymptomatic purpuric pigmentations developed on the flexure side of the right lower extremity in a linear arrangement 2-3 months before presentation. Histopathological analysis revealed focally perivascular inflammatatory infiltrates consisting mainly of lymphocytes and conspicuous extravasation of red blood cells into the papillary dermis. There was no histological evidence of vasculitis in either case. Hemosiderin deposits were more limited than expected in both patients. Both cases are compatible with diagnoses of pigmented purpuric dermatosis. Case 1 on the left arm was compatible with lichen purpuricus. Case 2 occurred in a linear arrangement on the right leg and had a history of aspirin use for chronic headache. We therefore suspected that medication had been an etiological or contributing factor.

FTS reduces bleomycin-induced cytokine and chemokine production and inhibits pulmonary fibrosis in mice.

Bleomycin (BLM), an antitumour drug, is known to cause interstitial pneumonia followed by pulmonary fibrosis, and has often been used to produce an animal model of pulmonary fibrosis. In the present study, we examined the effect of a nonapeptide thymic hormone, facteur thymique serique (FTS), on the murine lung fibrosis induced by intratracheal instillation of BLM. Treatment with FTS ameliorated BLM-induced fibrotic changes in a dose-dependent manner, as indicated by the reduced accumulation of hydroxyproline (HP). In addition, FTS suppressed BLM-induced cellular inflammatory response in the lungs, as evidenced by inhibition of increased lung weight, reduced accumulation of inflammatory leucocytes, including lymphocytes and neutrophils, but not macrophages, and less pronounced histopathological changes. Finally, BLM challenge increased the local synthesis of proinflammatory cytokines, TNF-alpha and IL-1beta and chemokines, MCP-1, MIP-1alpha RANTES, MIP-2 and KC, while administration of FTS suppressed the production of these cytokines, except for MCP-1. These effects of FTS were observed only when mice received intratracheal instillation with BLM. Considered collectively, our results indicated that FTS treatment ameliorated the cellular inflammatory responses and fibrotic changes in the lungs caused by BLM and such inhibition was well correlated with reduced synthesis of several fibrosis-related cytokines, and suggested that FTS may be potentially useful for the treatment of pulmonary fibrosis.

Candida albicans suppresses nitric oxide (NO) production by interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS)-stimulated murine peritoneal macrophages.

We examined the in vitro effect of Candida albicans on NO production by macrophages. Candida albicans suppressed not only NO production but also expression of inducible NO synthase (iNOS) mRNA by murine IFN-gamma and bacterial LPS-stimulated peritoneal macrophages. The suppression was not associated with inhibition but rather stimulation of IL-1 beta production. This effect was observed when more than 1 x 10(3)/ml of Candida albicans were added to macrophage cultures (1 x 10(6) cells/ml) and reached a maximal level at 1 x 10(6)/ml. The NO inhibitory effect of Candida albicans was mediated predominantly by as yet unidentified soluble factor(s) and to a lesser extent by direct contact. In addition, heat- or paraformaldehyde-killed Candida albicans did not show this inhibitory activity. Culture supernatant of Candida albicans also inhibited NO production by activated macrophages in a dose-dependent manner, and increased IL-1 beta production. Finally, the inhibitory effect was not mediated by IL-10 and transforming growth factor-beta (TGF-beta), since neutralizing antibodies to these cytokines did not influence Candida albicans-induced reduction in macrophage NO production. Our results suggest that Candida albicans may evade host defence mechanism(s) through a soluble factor-mediated suppression of NO production by stimulated macrophages, and that the effect is independent of production of immunosuppressive cytokines such as IL-10 and TGF-beta.

[Phase I-II study of 7-N-(P-hydroxyphenyl)-mitomycin C (KW2083, M83)].

A phase I-II study of KW2083, an analog of mitomycin C (MMC) was performed in a total of 22 patients. KW2083 was escalated by single intravenous administration of 40, 50, 60, 70, and 80 mg/m2 doses. Treatments were repeated every 4-6 weeks unless unacceptable toxicities occurred. The median times taken to reach nadir for each dose level were 9-12 days for leukocytes and 7-13 days for platelets respectively. The median times taken for recovery were 8-22 days for leukopenia and 10-37 days for thrombocytopenia. Variable and non-predictable hematological toxicity was observed in some cases. Biphasic hematological toxicity was observed in 4 courses. Acute toxicity occurred in 17 courses for 11 patients and consisted of nausea (44%), vomiting (13%), diarrhea (2.7%) and stomatitis (2.7%). Nephrotoxicity (elevation of BUN, 8.1% and proteinuria, 5.4%) occurred in 3 patients who had no previous impairment of renal function. Alopecia (8.1%) was also observed. Marked elevations of hepatic enzymes were noted in one patient who developed acute fulminant hepatitis with the second dose of KW2083. Objective response was observed in one of 20 evaluable patients. From this preliminary study, the maximum tolerable dose is 70 mg/m2 and the optimal dose of KW2083 was determined to be 50 mg/m2 at 6-week intervals. KW2083 has been introduced as an MMC analog of potential interest. However, hematological and non of hematological toxicities are very similar to those of MMC and does not appear that KW2083 will supersede MMC.

Phase I study of 7-N-(p-hydroxyphenyl)-mitomycin C.

The Phase I study of N-7-(p-hydroxyphenyl)-mitomycin C (KW 2083, M 83) was performed. The dose-limiting toxicity was leukopenia and thrombocytopenia and a maximum tolerable dose was 70 mg/m2. Nonhematologic toxicities included nausea (44%), vomiting (13%), diarrhea (2.7%), azotemia (8.1%), proteinuria (5.4%), alopecia (8.1%) and elevated hepatic enzymes (2.7%). This Phase I study indicates that the recommended starting dose for Phase II studies for patients without significant myelosuppression would be 50 mg/m2 at 6 week intervals in an intravenous push. KW 2083 should be avoided in patients with impaired renal functions and proteinuria because of permanent renal damages caused by the drug.

[The combination chemotherapy of vincristine, methotrexate, ACNU, and adriamycin for anaplastic carcinoma of the lung].

Recent advances in the chemotherapy of malignant diseases, particularly, in hematopoietic malignancies, has opened oncologists' eyes in wonder, whereas the chemotherapy of solid malignant diseases including the carcinoma of the lung is not satisfactory compared with the results of other modalities such as radiotherapy and surgery. The chemotherapy, however, gradually becomes a great importance because the majority of the cases of lung cancer is that of advanced one. Between June, 1974 and December 1980 we experienced 54 inoperable cases of lung cancers among which there were 11 cases diagnosed as an anaplastic carcinoma. The combination chemotherapy of vincristine (1 mg/body, iv, day 1), methotrexate (30 mg/body, iv, day 1 and 5), ACNU (100mg/body, iv, day 2) and adriamycin (40mg/m2, iv, day 2) was employed. Vincristine and methotrexate were given every 3 weeks and ACNU and adriamycin were repeated every 9 weeks. If the moderate degree of neuropathy due to vincristine occurred it was suspended and methotrexate was stopped if WBC was less than 2000/mm or if patients were suffered from stomatitis which disturbed their swallowing. According to the response criteria of Koyama-Saito 4, cases were responded and one of them survived 17 months after the initiation of above 4-drug combination chemotherapy, although she received another combination chemotherapy because of the relapse of disease. The combination chemotherapy of ACNU and adriamycin was tried to utilize the advantage of their time different effects on the bone marrow suppression and to cover heterogenous histopathological diagnosis of anaplastic carcinoma. The heterogeneity of anaplastic carcinoma included undifferentiated squamous cell carcinoma, adenocarcinoma, large cell carcinoma and even small cell carcinoma. In taking consideration of these points, the drug-combination was designed. Clinically, however, the long resting period made the tumor regrow in some cases due to severe delayed myelosuppression by the combination of ACNU and adriamycin. Thus, more cautiously-designed combination should be considered.

Phlebitis associated with the intravenous use of cephapirin and cephalothin in the combination therapy of antibiotics.

Phlebitis related to antibiotic infusion is one of the most frequent causes of morbidity in the debilitated patients with severe infection. There are a number of causes of infusion-induced phlebitis such as pH of intravenous fluid, needle used, and contamination of venipuncture site. Vein used to play an important role, particularly in patients with granulocytopenia receiving intravenous infusion. Cephalothin is an effective antibiotic in the treatment of granulocytopenic infection and is widely used currently. When cephalothin was introduced commercially, the frequency of phlebitis was as high as 50%. The main reason was thought to be acidity of the antibiotic solution. The cephalothin solution used currently is neutral in pH, but prevention of phlebitis is still not perfect. In contrast, cephapirin recently developed cephalosporin antibiotic, which resembles cephalothin in the antimicrobial activity and pharmacological properties caused less phlebitis than cephalothin in initial clinical studies. The patients receiving chemotherapy for malignant diseases frequently die of infections. A cephalosporin antibiotic is administered intravenously for a prolonged time in the presence of thrombocytopenia, and under such circumstances, other antibiotics such as carbenicillin (CBPC) and aminoglycoside are usually used in combination. The influence of these antibiotics injected through the same vein must be considered, but the possibility of phlebitis due to CBPC and aminoglycoside is negligible. In the present clinical study, 24 granulocytopenic patients were treated with the combination of antibiotics, cephapirin-carbenicillin-amikacin and cephalothin-carbenicillin-amikacin. Besides the clinical effect of the antibiotics, the incidence and severity of phlebitis were studied.

Clinical study of aclacinomycin A.

A phase I study of a new anthracycline antibiotic, aclacinomycin A, was performed in a total of 15 patients with advanced malignancy to determine the maximum-tolerated dose. The gastrointestinal toxicity which occurred was not dose-related and was not severe. Epilation and stomatitis were extremely minimal. Both hepatic dysfunction and hematologic toxicity were dose-limiting. A recommended dose for phase II study was determined to be 2.5--3.0 mg/kg (approximately 100--120 mg/m2) given in 3-week intervals. Objective response was observed in two patients with malignant lymphomas.

[Adrenal venography; improvement of the shape of catheter and its clinical application (author's transl)].

The preoperative identification of adrenal tumor is essential to correct diagnosis and appropriate treatment. Adrenal venography is one of the most useful adjuncts for this purpose, but catheters of various types introduced in the past are not necessarily well configured to catheterize adrenal veins. The catheter newly designed by us made catheterization of the left adrenal vein much easier even for unskilled hands. Our new catheter has been used in 20 of 55 venographical studies which have been carried out in our service. Determination of cortisol concentrations in adrenal venous blood which was performed in 6 cases failed to give any clue to determine the laterality of the existing adrenal tumor.