It is clinically important to diagnose diabetic neuropathy at the early stage. In this study, the toneentropy analysis of electrocardiogram was applied to diabetic patients to evaluate its usability in the screening of diabetic neuropathy. Consecutive 102 diabetic patients were invited to the study. Electrocardiogram was obtained and analyzed for the tone and the entropy using an original software developed previously. Nerve conduction velocity (NCV) was examined on the median, the posterior tibial and the sural nerves. Patients were divided into quartile subpopulations according to the NCVs in the analysis. Both the tone and the entropy significantly correlated with NCVs, while coefficient of variation in R-R intervals did not show a significant correlation. The correlation was most significant between the entropy and the NCV on the sural nerve. When a multivariate analysis (ordinary regression) was applied to examine independent effects of the factors influencing the NCV on the sural nerve, the entropy was the most potent independent factor (β = 1.14 ± 0.32, P = 0.0004) along with sex (β = 0.43 ± 0.19, P = 0.02) and BMI (β = 0.11 ± 0.05, P = 0.04). The tone-entropy analysis on electrocardiogram may be a promising non-invasive screening method for diabetic neuropathy.
Evaluation of immunosuppressive tumor-escape mechanisms in tumor-bearing hosts is of great importance for the development of an efficient tumor immunotherapy. We document here the functional characteristics of CD11b+Gr-1+ immature myeloid cells (ImC), which increase abnormally in tumor-bearing mice. Although it has been reported that ImC exhibit a strong immunosuppressive activity against T cell responses, we demonstrate that ImC derived from tumor-bearing mouse spleens (TB-SPL) did not exhibit a strong inhibitory activity against CTL generation in MLR. However, ImC isolated from TB-SPL and induced to differentiate into CD11b+Gr-1+F4/80+ uppressor macrophages (MΦ) under the influence of tumor-derived factors were immunosuppressive. Furthermore, we also demonstrate that ImC isolated from TB-SPL had a capability of differentiating into immunostimulatory dendritic cells (DC1) supportive of the generation of IFN-γ producing CTL if the ImC were cultured with Th1 cytokines plus GM-CSF and IL-3. Thus, our findings indicate that tumor bearing mouse-derived CD11b+Gr-1+ ImC are not committed to development into immunosuppressor cells but have dual differentiation ability into both immunosuppressive myeloid cells and immunos imulatory DC1.
We previously showed that sevoflurane anesthesia affected the expression ratios of 177 of 10,000 genes in multiple organs of rats by microarray analyses. The maximum number of altered genes was detected in the liver, and included several genes characterized as encoding drug-metabolizing enzymes (DMEs). Here, we investigated whether alterations of pharmacokinetic gene expressions after anesthesia differed between inhalation and intravenous anesthesia, and how long the alterations persisted after awakening from anesthesia. Livers were obtained from rats (n = 6 per group) anesthetized with sevoflurane, isoflurane, propofol or dexmedetomidine for 0 or 6 h, and rats awakened for 24 h after anesthesia for 6 h. The mRNA expression ratios of eight genes encoding DMEs that showed the greatest alterations in the previous study, namely Cyp7a1, Cyp2b15, Por, Nr1i2, Ces2, Ugt1a7, Abcb1a and Abcc2, were measured by quantitative real-time reverse transcriptase-polymerase chain reaction. The expression ratios were mostly increased after 6 h of anesthesia and returned to their control levels at 24 h after awakening from anesthesia. However, the expression ratios of some genes remained elevated for 24 h after awakening from anesthesia. There were differences between inhalation and intravenous anesthesia, and interestingly, between sevoflurane and isoflurane and between propofol and dexmedetomidine.
Normal rats were given catechin-rich green tea as drinking fluid and the effects on hepatic gene expression were examined. The results of DNA microarray analysis and quantitative real-time reverse transcription-polymerase chain reaction indicated the down-regulated expression of genes for glucose-6-phosphatase (G6Pase) and fatty acid synthase, and the up-regulated expression of peroxisome proliferator activated receptor α in the rats given green tea for 4 weeks as compared with the water-given animals. One may expect anti-diabetic activity by catechin-rich green tea through its chronic down-regulatory effect on G6Pase expression.
Vascular endothelial growth factor (VEGF) induces osteoclast differentiation as well as neovascularization by binding to the fms-like tyrosine kinase (Flt)-1 and fetal liver kinase (Flk)-1 receptors. The Flt-4 receptor also plays an important role in angiogenesis and lymphangiogenesis. The purpose of this study was to investigate the functions of Flt-4 in the signaling pathway of osteoclast differentiation. We examined the expression of Flt-4 on osteoclast precursor cells (OCPs), and the ability of recombinant human (rh) VEGF-D, one of the ligands of Flt-4, to stimulate the phosphorylation of extracellular-regulated kinase1/2 (ERK1/2) and to activate the nuclear factor-kappa B (NF-κB) pathway in OCPs. The number of osteoclasts induced by injection of rhVEGF-D in osteopetrotic (op/op) mice was also evaluated in the absence or presence of neutralizing antibodies to Flt-4. Flt-4 expression was detected on OCPs at both gene and protein levels and stimulation of Flt-4 by rhVEGF-D might induce activation of mitogen-activated protein kinase (MAPK) and NF-κB pathways for induction of osteoclast differentiation. Moreover, the number of osteoclasts in op/op mice increased after injection of rhVEGF-D, but was significantly reduced by the injection of Flt-4 neutralizing antibodies. We have therefore shown that Flt-4 expressed on OCPs, might activate MAPK and NF-κB pathways and played an important role in osteoclast differentiation.
The tibialis anterior and soleus muscles were fixed at the stretched or shortened positions to examine the influence of muscle length on muscle atrophy. Mice were divided into control (C), hindlimb suspension (HS), hindlimb suspension with ankle joint fixation at the maximum dorsiflexion (HSD), and hindlimb suspension with ankle joint fixation at the maximum plantarflexion (HSP). During the hindlimb suspension, the length of these muscles in the HS and HSP groups was very similar. Fourteen days after the hindlimb suspension, the atrophy of the tibialis anterior muscle in the HS and HSP groups was evidently milder than that in the HSD group, and that in the HS and HSP groups was very similar, suggesting that atrophy of the tibialis anterior muscle might largely depend on muscle length. Atrophy of the soleus muscle in the HSD group was milder than that in the HS and HSP groups, indicating that atrophy of the soleus muscle might also depend on muscle length. But atrophy of this muscle in the HSP group was milder than that in the HS group. These results demonstrate that some factors induced by the joint immobilization might be effective in preventing atrophy of the soleus muscle.
We established models of cancer-related anemia in mice from subcutaneous inoculation of two IL-6-producing cancer cell lines, human lung cancer cell line LC-06-JCK and murine colon26 clone 5 colon cancer cells. In both models, elevated levels of IL-6 were detected in sera and hemoglobin levels significantly decreased compared with non-tumor-bearing mice. In the LC-06-JCK model, serum albumin levels also decreased with elevated levels of human IL-6 in sera. On the other hand, serum levels of EPO increased, although anemia developed and did not improve. The development of cancer-related anemia was prevented by the administration of a rat anti-mouse IL-6 receptor antibody, MR16-1, in the LC-06-JCK model. It is therefore suggested that IL-6 causes anemia independent of a reduction in EPO levels. Our preclinical models should be useful for exploring new modalities for the treatment of cancer-related anemia.
Our previous study demonstrated that the pT2 and pT3-4 gallbladder carcinomas can be classified into two groups, i.e. infiltrative growth type (IG type) and destructive growth type (DG type) and that the DG type is associated with poor differentiation, aggressive infiltration, and decreased postoperative survival. The present study focused on the clinicopathologic significance of laminin-5γ2 chain expression as an indicator of local aggressiveness and Ki-67 labeling index (Ki-67 LI) as an indicator of the cell proliferation activity of gallbladder carcinoma. Ki-67 LI was higher in the DG type (26.3%) than in the IG type (21.4%), and the rate of high-grade cell proliferation cases (Ki-67 LI >= 30%) was high in the DG type (P = 0.012). Gallbladder carcinoma cases with high Ki-67 LI were significantly associated with poorly differentiation (P = 0.089) and distant lymph node metastasis (P = 0.079). Laminin-5γ2 expression patterns of gallbladder carcinoma were divided into two distinct types, extracellular staining and cytoplasmic staining. The extracellular staining was subclassified into two groups, basement membrane staining and stromal staining. In the basement membrane staining, laminin-5γ2 was present in the basement membranes surrounding neoplastic glandular structures. The basement membrane staining of laminin-5γ2 was more frequent in the IG type (40%) than in the DG type (12.9%) (P = 0.025). The stromal staining was more frequent in the DG type. Furthermore, the stroma-positive group was more closely associated with decreased overall survival than the stroma-negative group (P = 0.028). The cytoplasmic staining was not significantly correlated with invasion pattern in gallbladder carcinoma (P = 0.545). Univariate analysis demonstrated that laminin-5γ2 stromal staining is a predictor of lymphatic invasion, venous invasion, neural invasion, the mode of subserosal infiltration, and lymph nodal status. Multivariate analysis revealed the mode of subserosal infiltration is the strongest predictor of stromal invasion (P = 0.068). In conclusion, high-grade cell proliferation and stromal laminin-5γ2 staining were significan ly correlated with a wall-invasion pattern of aggressive gallbladder carcinoma indicating destructive growth (DG type).
Little is known regarding the association between the level of hyaluronic acid (HA) in saliva and dry mouth status. The aim of this study was to evaluate the salivary levels of HA in female patients with dry mouth (perceived xerostomia and hyposalivation) and compare them with agematched controls. We studied 46 females, and classified them into two groups based on perceived xerostomia and salivary flow rate, as well as a control group without symptoms. HA concentrations in unstimulated whole saliva were determined and a significant difference was found between the groups. The statistical association was stronger in patients (perceived xerostomia, normosalivation) administrated xerogenic drugs, while the HA levels in that group were significantly lower than those in the controls when converted to absolute amount of saliva per min. Within the limitations of the present study, patients with dry mouth had lower HA levels in saliva, which may serve as a marker of local dryness or oral mucosa lubrication.
Since many top swimmers wearing Speedo LZR Racer swimsuits have broken world records, it is considered that the corset-like grip of suit supports the swimmers to maintain flexibility of movement and reducing water resistance. We propose an alternative mechanism to explain this phenomenon. The suits are so tight that the blood circulation of swimmers is suppressed. This effect accelerates the anaerobic glycolysis system but rather suppresses the aerobic mitochondrial respiration system. Because of the prompt production of ATP in the glycolysis system, the swimmers, especially in short distance competitions, obtain instantaneous force in white fibers of the skeletal muscles.