Preventive effects of kilohertz frequency electrical stimulation on sepsis-induced muscle atrophy.

OBJECTIVES: The present study sought to evaluate the effect of electrical stimulation (ES) by using kilohertz frequency on muscle atrophy induced by sepsis. METHODS: Seventeen male ICR mice were randomly divided into 3 groups: control, lipopolysaccharide (LPS)-injected for 4 days, LPS plus ES (LPS+ES). Sepsis was induced by 4 days of an intraperitoneal LPS injection (10 µg/g body weight/day). LPS+ES animals received the LPS injections and ES twice a day for 4 days. ELISA and western blot analysis determined the plasma levels of inflammatory cytokines and ubiquitinated proteins, while the tibialis anterior muscles were weighed and muscle fiber cross-sectional area (CSA) were measured to assess muscle atrophy, which were analyzed by Student's t-test and ANOVA. RESULTS: LPS induced increased plasma levels of inflammatory cytokines, significant muscle mass loss (LPS: -29.0%, LPS+ES: -23.1%), decreased fiber cross-sectional area, and an up-regulation of atrogin-1 and ubiquitinated proteins in the tibialis anterior muscle compared with the control. ES attenuated the sepsis-induced loss of muscle mass and decreased fiber CSA, as well as attenuated the atrogin-1 and ubiquitinated protein up-regulation. CONCLUSIONS: Electrical stimulation may prevent sepsis-induced muscle atrophy through ubiquitin-proteasome pathway inhibition.

Abnormal asymmetries in subcortical brain volume in schizophrenia.

Subcortical structures, which include the basal ganglia and parts of the limbic system, have key roles in learning, motor control and emotion, but also contribute to higher-order executive functions. Prior studies have reported volumetric alterations in subcortical regions in schizophrenia. Reported results have sometimes been heterogeneous, and few large-scale investigations have been conducted. Moreover, few large-scale studies have assessed asymmetries of subcortical volumes in schizophrenia. Here, as a work completely independent of a study performed by the ENIGMA consortium, we conducted a large-scale multisite study of subcortical volumetric differences between patients with schizophrenia and controls. We also explored the laterality of subcortical regions to identify characteristic similarities and differences between them. T1-weighted images from 1680 healthy individuals and 884 patients with schizophrenia, obtained with 15 imaging protocols at 11 sites, were processed with FreeSurfer. Group differences were calculated for each protocol and meta-analyzed. Compared with controls, patients with schizophrenia demonstrated smaller bilateral hippocampus, amygdala, thalamus and accumbens volumes as well as intracranial volume, but larger bilateral caudate, putamen, pallidum and lateral ventricle volumes. We replicated the rank order of effect sizes for subcortical volumetric changes in schizophrenia reported by the ENIGMA consortium. Further, we revealed leftward asymmetry for thalamus, lateral ventricle, caudate and putamen volumes, and rightward asymmetry for amygdala and hippocampal volumes in both controls and patients with schizophrenia. Also, we demonstrated a schizophrenia-specific leftward asymmetry for pallidum volume. These findings suggest the possibility of aberrant laterality in neural pathways and connectivity patterns related to the pallidum in schizophrenia.

Two patients treated with simeprevir plus pegylated-interferon and ribavirin triple therapy for recurrent hepatitis C after living donor liver transplantation: case report.

We previously reported our data on telaprevir (TVR) used in combination with pegylated-interferon and ribavirin (PEG-IFN/RBV) for the treatment of recurrent hepatitis C virus (HCV) genotype 1 infection after liver transplantation (LT). TVR substantially increases the blood levels of immunosuppressive agents such as cyclosporine and tacrolimus for drug-drug interactions. On the other hand, the effect of simeprevir (SMV) on the blood levels of these immunosuppressive agents is unclear. We report 2 patients who achieved viral responses with little effect on the blood levels of cyclosporine and tacrolimus using SMV plus PEG-IFN/RBV treatment. The first was a 71-year-old woman with HCV-related liver cirrhosis and hepatocellular carcinoma who failed to respond to PEG-IFN/RBV after living donor LT. She was treated with 40 mg/d of cyclosporine, and received SMV plus PEG-IFN/RBV treatment. The second was a 65-year-old man with HCV-related liver cirrhosis who failed to respond to PEG-IFN/RBV after living donor LT. He was treated with 3 mg/d of tacrolimus, and received SMV plus PEG-IFN/RBV treatment. Serum HCV RNA became undetectable using TaqMan polymerase chain reaction (PCR) test after 4 weeks of treatment in both patients, and no remarkable fluctuation in blood concentration was observed either in cyclosporine or tacrolimus during the 12 weeks of SMV treatment. Completion of 12-week SMV triple therapy was followed by PEG-IFNα2b plus RBV, and both patients achieved sustained virological response 12 weeks after the end of treatment. SMV plus PEG-IFNRBV treatment showed a remarkable viral response with little effect on blood levels of immunosuppressive agents for recurrent HCV genotype 1 infection after LT.

TRPM8 has a key role in experimental colitis-induced visceral hyperalgesia in mice.

BACKGROUND: Transient receptor potential channel melastatin 8 (TRPM8) is activated by cold temperatures and cooling agents (menthol and icilin). Recent studies showed TRPM8 is expressed in visceral organs and peripheral sensory pathways. However, the role of TRPM8 in visceral hyperalgesia is poorly understood in pathological states such as inflammatory bowel disease. Hence, we investigated the distribution of TRPM8 and its involvement in visceral hyperalgesia in experimental colitis mice. METHODS: TRPM8 immunoreactivity was detected using immunohistochemical staining with fluorescein-conjugated tyramide amplification. Visceral hyperalgesia was measured by the intracolonic administration of TRPM8 agonist, WS-12, in control and dextran sodium sulfate (DSS)-induced colitis mice. KEY RESULTS: TRPM8 immunoreactivity in the distal colon was much higher than in the transverse and proximal colon under physiological conditions. TRPM8 immunoreactivity markedly increased in the distal colon mucosa of DSS-induced colitis mice compared with control mice. The number of TRPM8 nerve fibers in mucosa of DSS- or 2,4,6-trinitrobenzene sulfonic acid-induced colitis model mice drastically increased compared with control mice. TRPM8 immunoreactivities colocalized with the calcitonin gene-related peptide- and substance P-immunoreactive nerve fibers in the mucosa. Intracolonic administration of WS-12 induced behavioral visceral pain-like responses. The numbers of these responses in the colitis model mice were 3 times higher than in control mice, and were decreased by pretreatment with the TRPM8 channel blocker AMTB. CONCLUSIONS & INFERENCES: Increased expression of TRPM8 may contribute to the visceral hyperalgesia of experimental colitis.

Different expression patterns of hepatic cytochrome P450 s during anaphylactic or lipopolysaccharide-induced inflammation.

Certain physiological states and diseases can alter the expression and activity of cytochrome P450 s (CYPs), which have the potential to cause unexpected adverse effects. We previously demonstrated that lipopolysaccharide (LPS)-induced inflammation attenuates the induction of CYPs by xenobiotics in mouse liver. In this study, to investigate whether anaphylaxis-induced inflammation affects the hepatic CYPs' expression, we examined the effects of ovalbumin (OVA)-induced anaphylaxis on constitutive CYP mRNA and protein expressions. We also compared these effects with those obtained with LPS treatment. In addition, we examined the tumor necrosis factor (TNF) alpha and interleukin (IL)-113 mRNA levels, because these cytokines are known to be induced by LPS treatment and anaphylactic reactions. LPS treatment decreased the constitutively expressed Cyp1a2, Cyp2c29, and Cyp3al 1 mRNAs, and increased the TNFalpha and IL-1beta mRNAs. LPS treatment also decreased the CYP1A2 and CYP3A protein levels. Anaphylaxis, on the other hand, did not change the levels of the constitutively expressed Cyp1a2, Cyp2c29, or Cyp3a1 1 mRNAs, although it increased the TNFalpha and IL-1beta mRNAs, as observed in the LPS-treated mice. These results suggest that anaphylaxis-induced inflammation had less effect than LPS-induced inflammation on these CYPs in the liver. In contrast, we observed that the expressions of Cyp2b10 mRNA and its protein were quite different from those of the other CYPs in both the anaphylactic and LPS-treated mice. Our findings strongly suggest that the alteration of the constitutive CYPs' expression levels during inflammation varies according to the immunostimulation pathway.

Preventive effect of nucleoprotein on hindlimb unloading-induced capillary regression in rat soleus muscle.

We investigated the preventive effects of nucleoprotein on capillary regression and mitochondrial dysfunction induced by unloading in the soleus muscle of rats. Nucleoprotein is a supplement made from soft roe of salmon, and its major components are nucleotides and protamine. Adult male Sprague-Dawley rats were divided randomly into control, hindlimb unloading (HU), and hindlimb unloading plus nucleoprotein administration (HU+ NP) groups. Hindlimb unloading was carried out for 2 weeks in the rats belonging to the HU and the HU+ NP groups. The rats of the HU+ NP group were administered nucleoprotein (500 mg/kg) using a feeding needle twice a day for 2 weeks. Hindlimb unloading resulted in capillary regression, decreased succinate dehydrogenase activity of the muscle fiber, and decreased PGC-1α expression in the soleus muscle. These effects were prevented by administration of nucleoprotein. Nucleoprotein appears to prevent capillary regression and mitochondrial dysfunction caused by unloading of the skeletal muscle. Therefore, nucleoprotein supplementation may be an effective therapy for maintaining capillary network and mitochondrial metabolism of the muscle fiber during an unloading period.

Protective effects of astaxanthin on capillary regression in atrophied soleus muscle of rats.

AIM: The capillary regression in skeletal muscles associated with a chronic decrease in activity is related to a dysfunction of endocapillary cells induced by over-expression of oxidative stress. We hypothesized that treatment with astaxanthin, an antioxidant, would attenuate the oxidative stress induced by decreased skeletal muscle use, and that this attenuation would prevent the associated capillary regression. The purpose of the present study was to investigate the antioxidant and preventive effects of astaxanthin on capillary regression in the soleus muscle during hindlimb unloading. METHODS: Twenty-four adult male Wistar rats were assigned randomly either to a control, control plus astaxanthin treatment, hindlimb unloaded or hindlimb unloaded plus astaxanthin treatment group for 7 days. RESULTS: Hindlimb unloading resulted in a decrease in mean soleus absolute weight, capillary number, volume and luminal diameter. The accumulation of reactive oxygen species and the over-expression of superoxide dismutase (SOD-1), a decrease in the levels of vascular endothelial growth factor (VEGF) and its receptors, an inhibition of the angiopoietin pathway and an increase of thrombospondin-1 (TSP-1), as an anti-angiogenic factor were showed. Administration of astaxanthin attenuated the changes in SOD-1 and VEGF, up-regulated the angiogenic factors and reduced the capillary regression in the soleus of hindlimb unloaded rats. In addition, the VEGF-to-TSP1 ratio was higher in the astaxanthin treated groups than in the control and HU groups. CONCLUSION: These results suggest that astaxanthin may be an effective treatment to counter the detrimental effects of a chronic decrease in skeletal muscle use on the capillary network and associated angiogenic pathways.

Effects of exposure to hyperbaric oxygen on oxidative stress in rats with type II collagen-induced arthritis.

Arthritis was induced in 9-week-old female Dark Agouti rats by injecting type II collagen. Serum levels of the derivatives of reactive oxygen metabolites (dROMs), which are oxidative stress markers, and C-reactive protein (CRP) in arthritic rats that were exposed to a pressure of 1.25 atmospheres absolute and an oxygen concentration of 36% for 3 weeks (arthritis + HBO group) were compared to those of control rats (control group) and arthritic rats that were not exposed to hyperbaric oxygen (arthritis group). The body weights of the arthritis and arthritis + HBO groups were lower than that of the control group, whereas no difference in the body weight was observed between the arthritis and arthritis + HBO groups. The serum levels of dROMs and CRP in the arthritis group were higher than those in the control and arthritis + HBO groups. No difference in the serum level of CRP was observed between the control and arthritis + HBO groups. These results indicate that the conditions of hyperbaric oxygen exposure used in this study are effective for reducing the levels of reactive oxygen species, which are overproduced during arthritis.

The key role of stem cell factor/KIT signaling in the proliferation of blast cells from Down syndrome-related leukemia.

Transient leukemia (TL) has been observed in approximately 10% of newborn infants with Down syndrome (DS). Although treatment with cytarabine is effective in high-risk TL cases, approximately 20% of severe patients still suffer early death. In this study, we demonstrate abundant KIT expression in all 13 patients with GATA1 mutations, although no significant difference in expression levels was observed between TL and acute myeloid leukemia. Stem cell factor (SCF) stimulated the proliferation of the TL cells from five patients and treatment with the tyrosine kinase inhibitor imatinib suppressed the proliferation effectively in vitro. To investigate the signal cascade, we established the first SCF-dependent, DS-related acute megakaryoblastic leukemia cell line, KPAM1. Withdrawal of SCF or treatment with imatinib induced apoptosis of KPAM1 cells. SCF activated the RAS/MAPK and PI3K/AKT pathways, followed by downregulation of the pro-apoptotic factor BIM and upregulation of the anti-apoptotic factor MCL1. Although we found novel missense mutations of KIT in 2 of 14 TL patients, neither mutation led to KIT activation and neither reduced the cytotoxic effects of imatinib. These results suggest the essential role of SCF/KIT signaling in the proliferation of DS-related leukemia and the possibility of therapeutic benefits of imatinib for TL patients.

Microsurgery for vestibular schwannoma after gamma knife radiosurgery.

BACKGROUND: We evaluated the clinical characteristics of microsurgery for vestibular schwannoma (VS) after failed gamma knife radiosurgery (GKS). METHOD: Twelve patients, 5 men and 7 women aged 19 to 70 years (mean 54.5 years), who underwent microsurgery after failed GKS for VS were studied retrospectively. FINDINGS: The median interval between GKS and microsurgery was 28.8 months (range, 6.6-120 months) and 4 patients had undergone previous microsurgery. The mean volume of tumour at GKS was 6.9 cm(3) (range, 0.5-19.7 cm(3)) and the mean prescription dose to the tumour margin was 12.3 Gy. Microsurgery involved the lateral suboccipital approach in all patients. Tumour expansion involved solid enlargement in 7 patients, cystic enlargement in 3, and central necrosis in 2. Bleeding was slight in all patients except in one, probably because of the previous irradiation. Adhesion to the brain stem was severe in 7 patients. Identification of the facial nerve was easy in 5 operations and difficult in 7. Dissection of the tumour from the facial nerve was difficult in most interventions because of severe adhesions or colour change. Severe adhesions between the trigeminal nerve and the tumour was observed in 2 patients. The tumour was subtotally removed except around the internal auditory canal in most patients. Only one residual tumour increased in size and needed second GKS. The function of the facial nerve deteriorated in 3 patients, was unchanged in 7, and improved in 2. All patients had lost hearing on the affected side at the time of microsurgery. CONCLUSIONS: Microsurgery for VS after failed GKS presents some technical difficulties. Dissection of the tumour from the facial nerve or brain stem is likely to be difficult. We recommend subtotal resection without dissection of the facial nerve and tumour, because growth of the residual tumour was rare in our series.

Efficacy of gamma knife surgery for control of peritumoral oedema associated with metastatic brain tumours.

OBJECTIVES: To evaluate the efficacy of gamma knife surgery (GKS) for the control of peritumoral oedema associated with metastatic brain tumours. METHODS: A retrospective study of 280 consecutive metastatic brain tumours-100 from lung cancers, 100 from breast cancers and 80 from renal-cell carcinomas, associated with peritumoral oedema. The peritumoral oedema index was measured as A*B*C, where A (cm) was the maximum diameter of peritumoral oedema on the axial image, B (cm) was the maximum diameter perpendicular to A, and C (cm) was the maximum diameter on the coronal image. RESULTS: The oedema index of the renal cancer metastases was significantly larger than those of lung and breast cancer metastases (p<0.01). The oedema index of the renal cancer metastases at final imaging was also larger than those of lung (p<0.05) and breast (p<0.01) cancer metastases. The delivered marginal dose (22 Gy or more) was significantly correlated with tumour growth control by multivariate analysis (p = 0.03). Primary site (renal or not renal: p<0.01) and delivered marginal dose (25Gy or more: p = 0.04) were significantly correlated with control of peritumoral oedema by multivariate analysis. CONCLUSIONS: Brain oedema around metastatic brain tumours from renal-cell carcinomas was more extensive at the time of GKS and at final imaging compared with lung and breast cancer metastases. This paper suggests that the optimal doses for tumour growth control and brain oedema control may differ for metastatic brain tumours from renal-cell carcinomas.

Lack of exercise, via hindlimb suspension, impedes endogenous neurogenesis.

Bedridden patients who receive good physical rehabilitation are able to exhibit clinical improvement. Accumulating evidence demonstrates that exercise increases endogenous neurogenesis and may even protect against central nervous system (CNS) disorders. Here, we explored the effects of lack of exercise on neurogenesis in rats by employing a routine hindlimb suspension (HS) model over a 2-week period, which consists of elevating their tails, thereby raising their hindlimbs above the ground and unloading the weights in these extremities. In addition, the effects of exercise and recovery time with normal caging after HS were also explored. BrdU (50 mg/kg, i.p.) was injected every 8 h over the last 4 days of each paradigm to label proliferative cells. Immunohistochemical results revealed that HS significantly reduced the number of BrdU/Doublecortin double-positive cells in the subventricular zone and dentate gyrus. Exercise and recovery time significantly improved atrophy of the soleus muscle, but did not attenuate the HS-induced decrement in BrdU/Dcx-positive cells. A separate cohort of animals was exposed to the same HS paradigm and enzyme-linked immunosorbent assay (ELISA) of neurotrophic factors was performed on brain tissue samples harvested at the end of the HS period, as well as plasma samples from all animals. ELISA results revealed that HS reduced the levels of brain-derived neurotrophic factor in the hippocampus and vascular endothelial growth factor plasma levels. This study revealed that lack of exercise reduced neurogenesis with downregulation of neurotrophic factors. The use of the HS model in conjunction with CNS disease models should further elucidate the role of exercise in neurogenesis and neurotrophic factors in neurologic disorders.

Gamma knife radiosurgery for intracranial haemangioblastomas.

BACKGROUND: The results of gamma knife radiosurgery for haemangioblastomas were retrospectively studied to assess the efficacy for tumour growth control and clarify the clinical indications for gamma knife radiosurgery in these tumours. METHODS: The medical records of 22 patients with 67 tumours, 12 men and 10 women aged 20-73 years (mean 51.9 years), who underwent gamma knife radiosurgery for haemangioblastomas between January 1993 and January 2006, were retrospectively reviewed. Ten patients with 54 lesions had von Hippel-Lindau disease. The mean tumour volume was 1.69 cm(3) (range 0.0097-16.4 cm(3)). Nineteen patients had undergone 1-4 open surgery procedures (mean 1.5) before gamma knife radiosurgery. Tumours without a cystic component, (the solid type), were found in 54 lesions and tumours associated with cyst, (the mural nodule with cyst type), in 13 lesions. The marginal dose was 8-30 Gy (mean 14.0 Gy). FINDINGS: Follow-up magnetic resonance (MR) imaging was performed at 9-146 months (mean 63 months). The control rate for tumour growth was 83.6%. The only factor affecting tumour growth control was the presence of a cystic component at the time of gamma knife radiosurgery in both univariate and multivariate analysis. No complication such as radiation-induced peritumoural oedema or radiation necrosis occurred. CONCLUSION: The presence of cystic components at the time of gamma knife radiosurgery was the only factor significantly correlated with unfavourable tumour growth control by gamma knife radiosurgery for haemangioblastomas. Gamma knife radiosurgery is effective for solid type tumours, even if the marginal dose is relatively low. Surgical removal is recommended for mural nodule with cyst type tumours, when possible.

SLCO1B1 (OATP1B1, an uptake transporter) and ABCG2 (BCRP, an efflux transporter) variant alleles and pharmacokinetics of pitavastatin in healthy volunteers.

To investigate the contribution of genetic polymorphisms of SLCO1B1 and ABCG2 to the pharmacokinetics of a dual substrate, pitavastatin, 2 mg of pitavastatin was administered to 38 healthy volunteers and pharmacokinetic parameters were compared among the following groups: 421C/C(*)1b/(*)1b (group 1), 421C/C(*)1b/(*)15 (group 2), 421C/C(*)15/(*)15 and 421C/A(*)15/(*)15 (group 3), 421C/A(*)1b/(*)1b (group 4), 421A/A(*)1b/(*)1b (group 5), and 421C/A(*)1b/(*)15 (group 6). In SLCO1B1, pitavastatin area under plasma concentration-time curve from 0 to 24 h (AUC(0-24)) for groups 1, 2, and 3 was 81.1+/-18.1, 144+/-32, and 250+/-57 ng h/ml, respectively, with significant differences among all three groups. In contrast to SLCO1B1, AUC(0-24) in groups 1, 4, and 5 was 81.1+/-18.1, 96.7+/-35.4, and 78.2+/-8.2 ng h/ml, respectively. Although the SLCO1B1 polymorphism was found to have a significant effect on the pharmacokinetics of pitavastatin, a nonsynonymous ABCG2 variant, 421C>A, did not appear to be associated with the altered pharmacokinetics of pitavastatin.

Prediction of gait outcome with the knee-ankle-foot orthosis with medial hip joint in patients with spinal cord injuries: a study using recursive partitioning analysis.

STUDY DESIGN: Retrospective study of the degree of gait independence achieved by persons with spinal cord injury (SCI) using knee-ankle-foot orthosis with a medial single hip joint (MSH-KAFO). OBJECTIVE: To examine the effects of the neurological level, degree of paresis, age, and inhibitory physical/other factors on the gait with a MSH-KAFO in patients with SCIs. SETTING: Three university hospitals and two rehabilitation hospitals in Japan. METHODS: The 45 patients (36 men, nine women) examined included 10 with injuries in the cervical cord between C6 and C8 (group C), 20 with injuries in the upper-middle thoracic cord between T4 and T10 (group UT), and 15 with injuries in the lower thoracic-lumbar cord between T12 and L1 (group TL). Mean age was 34.0 years (range 16-68 years). Of these patients, 13 used the Walkabout, four used the gear joint, and 28 used the Primewalk as the medial hip joint. Recursive partitioning, which predicted the final status of gait from the level, degree of paresis, age, and inhibitory factors, was performed, and a decision tree for gait was constructed. Inhibitory factors were spasticity, involuntary spasms or muscle contractions, pain, contracture, weakness of the upper extremities, and decreased motivation to perform gait exercise. The degree of gait independence was rated on the following five-point scale: outdoor independent gait (5 points), indoor independent gait (4 points), indoor supervised gait (3 points), indoor assisted gait (2 points), and gait within parallel bars (1 point). New branches were added to the decision tree for gait based on the clinical experience, thereby constructing a new decision tree. RESULTS: The coincident ratio between the value predicted on the basis of the decision tree of gait and the value actually observed was 53.3%. The coincident ratio between the value predicted on the basis of the modified decision tree of gait and the actually observed value was 68.9%. CONCLUSION: The results provide valuable information to medical teams that may assist prescription of gait orthoses.

CYP2C76-mediated species difference in drug metabolism: a comparison of pitavastatin metabolism between monkeys and humans.

The monkey is often used to predict metabolism of drugs in humans since it generally shows a metabolic pattern similar to humans. However, metabolic profiles different from humans are occasionally seen in monkeys for some drugs including pitavastatin. Recently, we have successfully identified a monkey-specific cytochrome P450 (CYP) 2C76, which possibly accounts for a species difference between monkeys and humans because of its sequence and functional uniqueness. The present study on the role of CYP2C76 and other monkey CYP2Cs in pitavastatin metabolism, as an example, has revealed that CYP2C76 is important for the metabolism of the lactone form, indicating a major role of CYP2C76 for the difference in the metabolism of pitavastatin and possibly other drugs between monkeys and humans. The current investigation on the involvement of CYP2C76 in the metabolism of other drugs is expected to reveal further the further importance of this monkey-specific drug-metabolizing enzyme.

The balance of VEGF-C and VEGFR-3 mRNA is a predictor of lymph node metastasis in non-small cell lung cancer.

A positive association between vascular endothelial growth factor-C (VEGF-C) expression and lymph node metastasis has been reported in several cancers. However, the relationship of VEGF-C and lymph node metastasis in some cancers, including non-small cell lung cancer (NSCLC), is controversial. We evaluated the VEGF-C and vascular endothelial growth factor receptor-3 (VEGFR-3) expression in NSCLC samples from patients who had undergone surgery between 1998 and 2002 using real-time quantitative RT-PCR and immunohistochemical staining. We failed to find a positive association between VEGF-C and VEGFR-3 mRNA expression and lymph node metastasis in NSCLC. An immunohistological study demonstrated that VEGF-C was expressed not only in cancer cells, but also in macrophages in NSCLC, and that VEGFR-3 was expressed in cancer cells, macrophages, type II pneumocytes and lymph vessels. The VEGF-C/VEGFR-3 ratio of the node-positive group was significantly higher than that of the node-negative group. Immunohistochemical staining showed that VEGFR-3 was mainly expressed in cancer cells. The immunoreactivity of VEGF-C and VEGFR-3 was roughly correlated to the mRNA levels of VEGF-C and VEGFR-3 in real-time PCR. VEGF-C mRNA alone has no positive association with lymph node metastasis in NSCLC. The VEGF-C/VEGFR-3 ratio was positively associated with lymph node metastasis in NSCLC. This suggests that VEGF-C promotes lymph node metastasis while being influenced by the strength of the VEGF-C autocrine loop, and the VEGF-C/VEGFR-3 ratio can be a useful predictor of lymph node metastasis in NSCLC.

Serious cardiac and pulmonary calcification in a young peritoneal dialysis patient: potential role of continuous correction of acidosis.

We describe a 40-month-old male infant with renal failure, treated with peritoneal dialysis, who developed massive calcification of soft tissues including the heart and lungs with subsequent cardiopulmonary insufficiency. A diagnosis of Jeune syndrome was made. After starting peritoneal dialysis, the patient exhibited an intractable metabolic acidosis of unknown etiology necessitating treatment with intravenous or oral sodium bicarbonate. Myocardial calcification was first detected by 2-dimensional echocardiography performed 3 months after starting dialysis. The patient was not suitable for renal transplantation because of his cardiac dysfunction and died of cardiac and respiratory failure at the age of 6 years. Although the patient exhibited a variety of risk factors for ectopic calcification including hyperphosphatemia, hyperparathyroidism, high calcium-phosphate product and treatment with vitamin D, the early and massive soft tissue calcification may have been accelerated by correction of the metabolic acidosis. Therefore, the use of sodium bicarbonate may be involved in the etiology of the myocardial calcification.

Repeated gamma knife radiosurgery for multiple metastatic brain tumours.

BACKGROUND: The effectiveness of repeated gamma knife radiosurgery (GKS) for the treatment of multiple metastatic brain tumours was evaluated. METHODS: This study included 16 patients with 242 tumours, 10 men and 6 women with a mean age of 60.3 years at initial GKS, who underwent GKS four times or more for newly developed metastatic tumours. FINDINGS: Sixteen patients underwent a total of 83 GKS procedures (range 4 to 8, mean 5.2). The mean number of metastases at each GKS procedure was 2.9 and the number of tumours tended to increase at the 5th GKS procedure compared with the 1st, but not significantly. The mean interval between each procedure was 4.8 months and was not significantly different. Median survival was 22.4 months (range 9.4-78.9 months) and the primary site was not correlated with survival time. The total number of treated tumours tended to correlate to survival time, but not significantly. Use of adjuvant whole brain radiation also had no significant effect on survival time. The Karnofsky performance status was maintained at more than 70 in most patients, but decreased significantly between initial and final GKS. Death due to progression of brain lesions occurred in only about 30% of patients regardless of the multiple newly developed brain metastases. CONCLUSIONS: Repeated radiosurgery for brain metastases is effective and relatively long survival can be expected in some patients associated with a low risk of radiation-induced injury.

Betulinic acid augments the inhibitory effects of vincristine on growth and lung metastasis of B16F10 melanoma cells in mice.

We examined the antitumour effect of a combination of betulinic acid (BA) and vincristine (VCR) on murine melanoma B16F10 cells in vitro and in vivo. Betulinic acid, a pentacyclic triterpene, showed a synergistic cytotoxic effect on melanoma cells by combinational use of VCR. Betulinic acid and VCR induced cell cycle arrest at different points (BA at G1 phase and VCR at G2/M phase) and caused apoptosis in B16F10 melanoma cells. In the in vivo study, VCR inhibited metastasis of tumour cells to the lung. The addition of BA to VCR augmented suppression of the experimental lung metastasis of melanoma cells in C57BL/6 mice. The number of lung nodules of more than 1 mm in diameter in mice treated with BA and VCR was less than that in mice treated with VCR alone. These results suggest that BA is an effective supplement for enhancing the chemotherapeutic effect on malignant melanoma.