Research communications in molecular pathology and pharmacology最新文献

The effectiveness of pirfenidone ointment against thermoplasty-induced acute foreleg lameness in a double-blind study, and against acute and chronic lameness of musculoskeletal origin in an open multi-centered field trial was evaluated in this study. Thermoplasty was performed on both inner forelegs at designated locations of each horse under anesthetics. A 10% pirfenidone or placebo ointment was topically applied starting 24 hours after the thermoplasty three times daily for 7 days. For acute and chronic lameness of musculoskeletal origin, pirfenidone ointment was also applied one to three times daily for 7 to 10 days and continued for an additional 20 to 30 days. A marked swelling around the locations occurred in 24 hours post-thermoplasty. The topical application of pirfenidone ointment not only caused a significant reduction in the circumference measurements at 5, 6 and 7 days, but it also decreased the changes in the circumferences from pre-thermoplasty as an index of edema, at 3, 4, 5, 6, and 7 days when compared to the placebo ointment at the corresponding times. Although treatment for 7 days of acute leg lameness of musculoskeletal origin with topical pirfenidone ointment caused significant decreases in swelling, heat, and pain, and improved the degree of flexion when compared with the pretreatment, it had little effect on chronic lameness except that it improved the flexion at the second-exam interval. It was concluded that topical application of pirfenidone is effective for treatment of acute lameness resulting from thermoplasty or from various types of musculoskeletal disorders, suggesting that pirfenidone offers a promising therapeutic potential to manage acute inflammation, an important component of lameness.

Previous studies have shown that congenic bone marrow transplantation into apolipoprotein E-deficient mice prevented hypercholesterolemia and atherosclerosis. In this study, we examined the effect of intravenous administration of human umbilical cord blood (HUCB) mononuclear cells on the progression of atherosclerosis in male homozygous mice that had mutation in the low-density lipoprotein receptor (Ldlr(tm1Her) mutation mice). In addition, the effect of human breastmilk alone as well as the combination of HUCB cells and breastmilk was studied on the prevention of atherosclerosis in these mice. In all groups of mice, atherosclerosis was predominant in the ascending aorta, but the rest of the aorta had variable evidence of atherosclerosis. Treatment of mice with HUCB cells significantly ameliorated the development of atherosclerosis in the ascending aorta, as compared with untreated mice; whereas breastmilk alone did not have any significant effect. A similar beneficial effect was observed with the combination therapy, which could be attributed only to HUCB cell treatment. There was no beneficial effect of treatment on the thoracic and abdominal aorta. Thus, early administration of HUCB cells prevents the progression of atherosclerosis in the ascending aorta of mice that are prone to the development of atherosclerosis. This beneficial effect occurred without any immunosuppression and graft-vs-host disease.

Background/aims: Matrix metalloproteinases (MMPs) play important roles in cancer invasion and metastasis. Recently, functional gene polymorphisms in the promoter regions of MMP-1 1G/2G, MMP-3 5A/6A and MMP-9 C/T have been found, and have been reported to be associated with the prognosis of various cancers. In this study, we examined the association of the MMP-1, -3, -9 genotypes with the prognosis of hepatocellular carcinoma (HCC).

Methods: Ninety two patients with hepatitis C virus related HCC were enrolled in the study. Genomic DNA samples were extracted from peripheral white blood cells, and gene polymorphisms of MMP-1, MMP-3and MMP-9 were analyzed by PCR-RFLP methods.

Results: There were no significant differences of the survival rate between MMP-1 1G carriers and 2G homozygotes, and between MMP-9 T carriers and C homozygotes. By contrast, HCC patients with MMP-35A allele had a significantly poorer prognosis than 6A homozygotes.

Conclusions: Unlike MMP-1 and MMP-9 genotypes, MMP-3 5A allele, with higher transcriptional activity, may be a risk factor for the poor prognosis of HCC patients.

Administration of antitumor drugs such as 5-fluorouracil and methotrexate to rats causes intestinal injury. The intestinal injury was reported to be prevented by coadministration of prostaglandin E1 analogues to rats. IEC-6 cell is a useful in vitro model to study the cytoprotective effect in enterocytes, For its purpose, it is important to know whether or not prostaglandins are taken up into IEC-6 cells. The transport of prostaglandin E1 into IEC-6 cells was investigated, using [3H]-prostaglandin E1. Prostaglandin E1 was found to be transported into IEC-6 cells by the passive diffusion mechanism.

Unlabelled: Interferon (IFN) and especially IFN-alpha exhibit clinical anti-tumor activity against various types of malignant diseases. Natural inhibitors to various cytokines and IFNs have been documented in vitro as well as in vivo. IFN inhibitors have been implicated for the ineffectiveness of IFN treatment in malignant neoplasias. The aim of this study was to investigate the incidence of the IFN inhibiting activity in serum from patients with haematological malignancies versus patients with solid tumours, as an effort to explain, just in part, the different response of these patients to IFN treatment.

Patients and methods: Ninety patients with a clinically evident solid tumour and forty-six patients with haematological malignancies were included in the study. Serum samples from all patients were collected before any treatment and stored at -70 degrees until use. Controls sera were selected from 50 apparently healthy blood donors. Interferon-inhibiting activity as well as endogenous IFN-like activity were determined in all serum samples in a cell line highly sensitive to IFN.

Results: There was no endogenous IFN-like activity in any of the patients' group or controls' group. Sera from patients with haematological malignancies exhibited IFN-blocking activity at a lower percentage (21.7%) in comparison to sera from patients with solid tumours (56.6%, P<0.001), but at a significantly higher percentage in comparison to sera from controls (P<0.01).

Conclusions: The fact that IFN inhibitors were detected at a significantly lower percentage in sera from patients with haematological malignancies versus patients with solid tumours, could explain in part the better response of the haematological malignancies to IFN treatment.

This study aims to understand the effect of ten-week passive repetitive plyometric (PRP) training on human skeletal muscle and the application of PRP training for performance. Vastus lateralis of nine candidates were aspirated before (pre) and after (post) PRP training. Histochemical approaches with regular hematoxylene-eosin (HE) and Mallory's phosphotungstic acid hematoxylin (PTAH) stains were used to demonstrate the changes of muscle fibers. Immunohistochemical studies with heat shock protein (anti-hsp72, Stressgen, Canada) were employed to display cellular activities. Each set of slides was quantitatively analyzed by using a modified morphometric method (Russ and Dehoff, 1999) on a Nikon ECLIPSE 80i microscope, equipped with an Evolution VF COOLED color video camera, and the Image-Pro Plus software (5.0 for Win; Media Cybernetics, USA). Finally, hsp72 mRNAs of both pre-PRP and post-PRP specimens were amplified through RT-PCR. Signal intensities were read by a densitometer and analyzed through the SPSS (11.0 for Win) statistically. Post-PRP muscle cells demonstrated hypertrophic change with increased cellular content and a narrowed inter-cellular space according to both HE and PTAH profiles. Post-PRP cellular hsp72 proteins were higher by up to five percent, as measured by a gray-scale reading. Further, after a training period of 10 weeks, hsp72 mRNA expression was several times higher.

We have investigated the role of cyclooxygenase-2 (COX-2) in myocardial infarction (MI) and ischaemia in rabbits subjected to isoprenaline (ISP) a potent beta-adrenergic agonist. The acute phases of MI and repair mimicked those which occurred in humans. MI after induction with ISP was monitored by following increases seen in the levels of serum enzymes, Troponin I and Creatinine phosphokinase (CPK) in rabbits before and after ISP induced MI. Electrocardiographic (ECG) changes showed typical ST elevation and q-wave development. Histochemical studies of the rabbit heart on 2nd day after ISP injection showed changes of coagulation necrosis. Day 4 total coagulation with the loss of nuclear and striation associated with heavy interstitial infiltrate of neutrophils was found. Day 8 after infarction showed collagen deposition with capillary channels in between the remaining islands of myocytes in the infarcted area on the 16th day scarring was complete. Coronary perfusion rates (CPR) of the infarcted and nimesulide (a COX-2 inhibitor) treated rabbits displayed significant improvement on each corresponding day after infarction as compared to the infarcted and saline treated rabbits (P<0.01). These results suggest that nimesulide, a COX-2 inhibitor exhibit cardioprotective effects in MI.

N3-(2',5 '-Dimethoxyphenacyl)arabinofuranosyluracil (N3-(2',5 '-DiMeOPhAc)AraU) is a pyrimidine nucleoside derivative which possesses antinociceptive effect by intracerebroventricular (i.c.v.) administration to mice. The compound (100 microM) significantly decreased the specific binding of [3H]D-Ala2, N-Me-Phe4, Gly5-ol-enkephalin (DAMGO) and [3H]D-Pen2, D-Pen5-enkephalin (DPDPE) at the mu- and delta-opioid receptor, respectively, but has no effect on the binding of [3H]U-69,593 at the kappa-opioid receptor of rat brain slices. The autoradiographic studies also demonstrated that [3H]DAMGO and [3H]DPDPE bindings on the rat brain slice were displaced by N3-(2',5'-DiMeOPhAc)AraU. These results indicate that N3-(2',5'-DiMeOPhAc)AraU interacts with mu- and delta-opioid receptors.

Because isoniazid is a selective inducer of CYP2E1 and isoniazid-induced hepatotoxicity is believed to be due to activation of its metabolites by CYP450, this study was undertaken to determine the effect of isoniazid containing regimen on CYP2E1 in TB-patients. The activity of CYP2E1 in 11 newly diagnosed TB-patients (5 F, 6 M) was investigated before (day 0) and during (day 14) treatment for tuberculosis. CYP2E1 activity was measured using the plasma metabolic ratio (MR) of 6-hydroxy-chlorzoxazone to chlorzoxazone, while CYP2E1 quantity in the peripheral lymphocytes was measured using SDS-PAGE. By day 14 of anti-tuberculosis treatment, the activity of CYP2E1 was inhibited by 72% in 8 patients, but increased in 3 patients. The MR for the 8 patients was reduced from (Median & Range) 2.78 (1.1-21.5) on day 0, to 0.75 (0.4-1.22) on day 14, (P = 0.0006). Renal function was normal before and during the investigation. The detection of CYP2E1 by in peripheral lymphocytes was so variable that it could not be correlated with enzyme activity. Nevertheless, its detection in peripheral lymphocytes where normally is not resident indicates that CYP2E1 was induced by isoniazid. These results indicate that during treatment for tuberculosis with isoniazid containing regimen, CYP2E1 is induced but its activity is inhibited by isoniazid.

The FHIT gene, located at human chromosome 3p14.2 spanning the FRA3B common fragile region, is frequently altered in several types of human cancers. To study the potential role of FHIT gene in colorectal cancer, expression of the FHIT gene were examined from 20 colorectal cancers for by reverse transcription-polymerase chain reaction (RT-PCR) and all of aberrant transcripts were cloned and sequenced. In addition, the effect of exogeneous rat FHIT overexpression on cell cycle was investigated by introducing the gene into normal rat kidney cells (NRK-52E). In RT-PCR, 7 cases of 25 patients with colorectal cancer showed 16 transcripts of abnormal sizes. Sequence analysis of the abnormal transcripts revealed these transcripts due to the deletion of multiple entire exons or part of exon sequences by errors in the splicing of pre-mRNA. The inserts of 59-bp and 138-bp sizes occurred in combination with in-frame deletions and was identified as part of the FHIT intron 5 sequence. In cell cycle analysis, over-expression of the FHIT gene in the FHIT-pTARGET-transformed NRK-52E cells did not affect cell proliferation and cell cycle distribution. Taken together, although alternative splicing of human FHIT is not directly associated with carcinogenicity, FHIT is frequently inactivated by exon skipping, intron retention, and activation of cryptic splice site within exon 6 in colorectal cancer.