A Potential Serum N-glycan Biomarker for Hepatitis C Virus-Related Early-Stage Hepatocellular Carcinoma with Liver Cirrhosis.

Detection of early-stage hepatocellular carcinoma (HCC) is beneficial for prolonging patient survival. However, the serum markers currently used show limited ability to identify early-stage HCC. In this study, we explored human serum N-glycans as sensitive markers to diagnose HCC in patients with cirrhosis. Using a simplified fluorescence-labeled N-glycan preparation method, we examined non-sialylated and sialylated N-glycan profiles from 71 healthy controls and 111 patients with hepatitis and/or liver cirrhosis (LC) with or without HCC. We found that the level of serum N-glycan A2G1(6)FB, a biantennary N-glycan containing core fucose and bisecting GlcNAc residues, was significantly higher in hepatitis C virus (HCV)-infected cirrhotic patients with HCC than in those without HCC. In addition, A2G1(6)FB was detectable in HCV-infected patients with early-stage HCC and could be a more accurate marker than alpha-fetoprotein (AFP) or protein induced by vitamin K absence or antagonists-II (PIVKA-II). Moreover, there was no apparent correlation between the levels of A2G1(6)FB and those of AFP or PIVKA-II. Thus, simultaneous use of A2G1(6)FB and traditional biomarkers could improve the accuracy of HCC diagnosis in HCV-infected patients with LC, suggesting that A2G1(6)FB may be a reliable biomarker for early-stage HCC patients.

Insulin resistance derived from zinc deficiency in non-diabetic patients with chronic hepatitis C.

Chronic hepatitis C virus (HCV) infection frequently evokes metabolic abnormalities including insulin resistance. A decrease in serum zinc (Zn) levels is often observed in association with hepatic fibrosis. Zn also plays important roles in insulin secretion. However, little is known about the relationship between Zn deficiency and insulin resistance in patients with HCV-related chronic liver disease. The main purpose of this study was to examine the contribution of Zn deficiency to insulin resistance in patients with chronic hepatitis C (CH-C). Forty-eight non-diabetic patients with CH-C were enrolled. Serum alanine aminotransferase (ALT), ferritin and Zn levels were examined in the enrolled patients with CH-C. Insulin resistance was determined by the Homeostasis model for assessment of insulin resistance (HOMA-IR). Zn deficiency was defined as serum Zn levels <65 µg/dl. Seven out of the 48 (15%) patients with CH-C fulfilled the criteria for Zn deficiency. Serum Zn levels were inversely correlated with serum ferritin levels (r=-0.364, p=0.0140). The values of HOMA-IR were positively linked to serum ferritin levels (r=0.299, p=0.0484). The mean value of HOMA-IR in the Zn deficiency group was significantly higher than that in the normal-range Zn group (3.76±0.66 vs. 2.08±1.35, p=0.0019). Serum ALT levels were also closely associated with serum ferritin levels (r=0.727, p<0.001). These findings were independent of HCV genotypes or loads of HCV-RNA. Our data suggest that iron overload in patients with CH-C derives from Zn deficiency and thereby causes insulin resistance.

Paraneoplastic necrotizing myelopathy in a patient with advanced esophageal cancer: An autopsied case report.

We report the first autopsied case of paraneoplastic necrotizing myelopathy associated with esophageal cancer in the literature. The patient had acute flaccid paraplegia and urinary retention, and had a good recovery of strength of both legs in response to corticosteroids. MRI showed a characteristic lesion with post-gadolinium enhancement of the cervical to mid-thoracic spinal cord at the onset, which has never been reported. Taken together, these results suggest that there is an underlying autoimmune mechanism in paraneoplastic necrotizing myelopathy.

Clinical characteristics of patients with hepatitis C virus-related chronic liver disease seropositive for anticentromere antibody.

The association between anticentromere antibody (ACA) and hepatitis C virus (HCV) infection remains unclear. We subjected eight patients with HCV-related chronic liver disease (CLD) seropositive for ACA to a battery of clinical and laboratory tests. The patient cohort was dominated by females, and four of the eight (50%) patients had a concomitant autoimmune disease. All of the patients had high titers of ACA (>or=1:320). The histological activity index scores in chronic hepatitis C (CH-C) patients with ACA were significantly higher than those in CH-C patients without antinuclear antibody (ANA) (12.8 +/- 1.8 vs. 8.3 +/- 4.5, P = 0.0372). The frequency of human leukocyte antigen (HLA) DR-8 in patients with HCV-related CLD seropositive for ACA was significantly higher than that in patients with CH-C seronegative for ANA (71 vs. 18%, P = 0.0108). These findings suggest that ACA is induced by chronic HCV infection in association with HLA DR-8, and that CH-C patients with ACA exhibit more severe hepatic fibrosis and inflammation than CH-C patients without ANA.

Outer arm fucosylation of N-glycans increases in sera of hepatocellular carcinoma patients.

Serum glycans are promising markers for early-stage cancer detection, but the research remains challenging because low concentrations of serum glycoproteins are secreted from early-stage tumors. We have established an N-glycan profiling method using liquid chromatography electrospray ionization-mass spectrometry with high sensitive derivative, trimethyl(4-aminophenyl)ammonium chloride (TMAPA). The mass sensitivity of TMAPA-labeled oligosaccharides was enhanced more than 50 times compared with 2-aminopyridine (PA) labeled oligosaccharides, and the analytical period was significantly shortened compared with traditional HPLC 2D-mapping. Using this method, we found about 28 major N-linked oligosaccharides in human sera, and we investigated their alterations in patients who developed hepatocellular carcinoma (HCC). We found that outer arm fucosylation (attached GlcNAc via an alpha 1-3/4 linkage) in highly branched oligosaccharides increased significantly in sera of HCC patients. Normalizing the level of outer arm fucosylation by taking into account platelet concentration allowed us to distinguish more clearly between HCC and LC patients.

The efficacy of ursodeoxycholic acid and bezafibrate combination therapy for primary biliary cirrhosis: A prospective, multicenter study.

AIM: Treatment with ursodeoxycholic acid (UDCA) improves the survival of stage I and II primary biliary cirrhosis (PBC) patients. However, new therapeutic options are needed for patients who are refractory to UDCA and for those whose disease is at an advanced stage. Bezafibrate could be useful in PBC treatment, since it increases phospholipid output into the bile and reduces the cytotoxicity of hydrophobic bile acids, which are increased with cholestasis. METHODS: We conducted two prospective, multicenter randomized open studies in non-cirrhotic patients with PBC to evaluate the efficacy of bezafibrate. One study compared UDCA and bezafibrate monotherapy (study 1: 45 patients [37 females], mean age 55.9 years), and the other evaluated the addition of bezafibrate to patients who were refractory to UDCA (study 2: 21 patients [18 females], mean age 54.1 years). RESULTS: Study 1 demonstrated that bezafibrate monotherapy was as effective as UDCA and study 2 revealed that bezafibrate combined with UDCA was effective in improving and maintaining biliary enzymes where the ineffectiveness of long-term treatment with UDCA was confirmed. CONCLUSION: This multicenter, randomized, open study revealed that combination therapy of bezafibrate and UDCA improved biliary enzymes in non-cirrhotic Japanese patients with PBC refractory to UDCA. Further studies are needed to evaluate whether combination therapy improves histological staging and prognosis.

A combination therapy of ethanol injection and radiofrequency ablation under general anesthesia for the treatment of hepatocellular carcinoma.

AIM: To summarize the effects of laparoscopic ethanol injection and radiofrequency ablation (L-EI-RFA), thoracoscopic (T-EI-RFA) and open-surgery assisted EI-RFA (O-EI-RFA) under general anesthesia for the treatment of hepatocellular carcinoma (HCC). METHODS: Time-lag performance of RFA after ethanol injection (Time-lag PEI-RFA) was performed in all cases. The volume of coagulated necrosis and the applied energy for total and per unit volume coagulated necrosis were examined in the groups treated under general (group G) or local anesthesia (group L). RESULTS: The results showed that the total applied energy and the applied energy per unit volume of whole and marginal, coagulated necrosis were significantly larger in group G than those in the group L, resulting in a larger volume of coagulated necrosis in the group G. The rate of local tumor recurrence within one year was extremely low in group G. CONCLUSION: These results suggest that EI-RFA, under general anesthesia, may be effective for the treatment of HCC because a larger quantity of ethanol and energy could be applied during treatment under painfree condition for the patients.

The interaction between somatosensory and auditory cognitive processing assessed with event-related potentials.

Event-related potentials induced by somatosensory and after auditory stimulation or bilateral somatosensory stimulation in young adults were studied, aiming to evaluate interactions of cognitive processing of somatosensory and auditory information. In experiment 1, event-related potentials were elicited by four stimulus conditions which were randomly presented with an interstimulus interval of 20 +/- 3 milliseconds: electric stimuli delivered to the right median nerve (Con 1), auditory stimuli delivered to both ears (Con 2), electric and subsequent auditory stimuli (Con 3), and electric stimuli delivered to the bilateral median nerve (Con 4). In experiment 2, Con 2 was changed to electric stimulation of the left median nerve. The subtraction of grand averaged event-related potentials of Con 1 and Con 2 from those of Con 3 demonstrated a positive-negative peak complex with a positive peak at approximately 165 to 170 milliseconds after stimuli and a negative peak with a mean latency of 372 milliseconds in experiment 1. In experiment 2, the subtraction of event-related potentials in Con 1 and 2 from those in Con 4 revealed positive and negative peaks resembling those in experiment 1, but the distribution or latencies of the peaks differed from those in experiment 1. We speculate that the subtracted positive and negative peaks reflect different cognitive processing of bimodal and bifocal sensory information.

How many cytological examinations should be performed for the diagnosis of malignant biliary stricture via an endoscopic nasobiliary drainage tube?

BACKGROUND AND AIM: The sensitivity of bile cytology is recognized as being low. Repeating cytological sampling is likely to improve the sensitivity. The aim of this study is to determine the optimal number of repeated cytological sampling of bile obtained via an endoscopic nasobiliary drainage (ENBD) tube for the diagnosis of malignant biliary stricture. METHODS: Ninety-eight patients with malignant biliary stricture who underwent ENBD were enrolled. Diagnoses included bile duct carcinoma (n = 53), pancreatic carcinoma (n = 28), carcinoma of the major papilla (n = 8), gallbladder carcinoma (n = 6), and hepatocellular carcinoma (n = 3). Bile was aspirated via an ENBD tube once a day and immediately evaluated cytologically. RESULTS: The median number of cytological samplings via an ENBD tube was 2.8 times (range, 1-10). In 40 of 98 patients with malignant biliary stricture, cytology was positive at the first cytological sampling (sensitivity 40.8%). Cytology was cumulatively positive in 71 of 98 patients (sensitivity 72.4%) from which repeated samples were taken. In 71 patients with positive cytology, correlation of the positive rate and the number of cytological samplings performed was investigated. In 68 of 71 (95.8%) patients with positive cytology, positive results were obtained by or at the sixth examination. CONCLUSIONS: Bile cytology via an ENBD tube is an easy method, and has been shown to have relatively high sensitivity. The optimal number of repeated cytological samplings using bile obtained via an ENBD tube for the diagnosis of malignant biliary stricture was concluded to be six.

[A case of intrahepatic splenosis].

When a 32-year-old man was 18 a traffic accident caused splenic rupture, for which he underwent splenectomy, complicated postoperatively by hepatitis. As part of routine follow-up with his family doctor for chronic hepatitis C, a hepatic tumor was detected on plain abdominal computed tomography (CT), and he was admitted to our hospital for further investigation and treatment. Based on the results of abdominal dynamic CT, abdominal magnetic resonance imaging(MRI), abdominal angiography, CT during hepatic arteriography (CTHA) and CT during arterial portography (CTAP), we performed a laparotomy for suspected hepatocellular carcinoma. The operative findings were a 40 mm dark purple-brown nodule on the surface of the S7 segment of the liver, as well as multiple nodules, less than 20 mm in size but otherwise of similar appearance, scattered over the abdominal viscera and peritoneum, in particular the greater omentum. A hepatectomy was not performed, as we considered this to be a case of intrahepatic splenosis, so the abdomen was closed after a liver biopsy was taken. Histopathological examination of this specimen confirmed the diagnosis. As there have been only 19 previous reported cases of intrahepatic splenosis, we hereby present this valuable case with a review of the literature.

Comparative study of the effects of percutaneous ethanol injection and radiofrequency ablation in cases treated with a straight or expandable electrode.

Radiofrequency ablation (RFA) has become mainstream among non-surgical treatment modalities in clinical settings for the treatment of hepatocellular carcinoma. We have previously described the novel combination therapy of percutaneous ethanol injection and RFA (PEI-RFA) and reported that this combination therapy was more effective than RFA alone in terms of the induced volume of coagulated necrosis and the energy requirement for the treatment. RFA instruments are mainly divided into two types according to the electrode used, either the straight or expandable type electrode. Although PEI-RFA can be performed by either of the electrodes, there may be some important differences in PEI-RFA according to the type of electrode used. In the present study, the effect of using the straight or expandable electrode in PEI-RFA was evaluated by analyzing the ablation time, volume of coagulated necrosis, the energy requirement for ablation and the amount of injected ethanol into HCC. The comparative study showed that ablation time, total energy requirement and per unit volume of energy requirement for whole and marginal coagulated necrosis were significantly smaller in the group treated with the expandable electrode (E group) than those in the group treated with the straight electrode (S group). The volume of coagulated necrosis was similar between these groups. In group E, the amount of injected ethanol showed a positive correlation with the volume of coagulated necrosis and the size of the tumors. These results suggest that prior injection of ethanol works mainly by shortening the time and energy requirement for ablation in the time-lag PEI-RFA using the expandable electrode. Thus, prior injection of ethanol before RFA may make RFA treatment less invasive in the time-lag PEI-RFA using the expandable electrode as previously shown HCC cases treated with straight electrode.

Assessment of autonomic dysfunction of multiple system atrophy with laryngeal abductor paralysis as an early manifestation.

Laryngeal abductor palsy (LAP) is common in the advanced stages of multiple system atrophy (MSA). However, occurrence of LAP in the early stages might make a diagnosis of MSA difficult. To search for a clue to diagnosis of MSA with LAP as an early manifestation, we assessed the clinical features of autonomic dysfunction and the central cardiovascular control circuits in two MSA patients who had LAP as a cardinal symptom in the early stages. Development of autonomic dysfunction was preceded or followed by LAP. The autonomic symptom occurring predominantly in the earliest stages was urinary disturbance rather than orthostatic hypotension. Although screening cardiovascular autonomic function tests did not conclusively indicate a diagnosis of MSA, vasopressin release in response to head-up tilt and growth hormone response to clonidine administration demonstrated inappropriate responses, suggesting that the noradrenergic neurons of the caudal ventrolateral medulla were impaired. Diagnosis of atypical MSA with LAP in the early stages might be accelerated by a detailed investigation focused on urinary symptoms and neuroendocrine approaches.

Efficacy of zinc administration in patients with hepatitis C virus-related chronic liver disease.

OBJECTIVE: Zinc supplementation has been shown to contribute to inhibition of liver fibrosis and improvement in hepatic encephalopathy. However, little is known about the anti-inflammatory effect of zinc on hepatitis C virus (HCV)-related chronic liver disease (CLD). We therefore examined the effects of zinc administration on inflammatory activity and fibrosis in the liver of patients with HCV-related CLD. MATERIAL AND METHODS: Polaprezinc, a complex of zinc and l-carnosine, was administrated at 225 mg/day for 6 months to 14 patients with HCV-related CLD, in addition to their ongoing prescriptions. Peripheral blood cell counts, liver-related biochemical parameters, serological markers for liver fibrosis, HCV-RNA loads, and serum levels of zinc and ferritin were evaluated before and after zinc administration. RESULTS: Serum zinc concentrations were positively correlated with hepatic reserve before zinc supplementation. A significant increase in serum zinc level was observed after zinc supplementation (64+/-15 versus 78+/-26 mg/dl, p=0.0156). Treatment with polaprezinc significantly decreased serum aminotransferase levels (aspartate aminotransferase (AST): 92+/-33 versus 63+/-23 IU/l, p=0.0004; alanine aminotransferase (ALT): 106+/-43 versus 65+/-32 IU/l, p=0.0002), whereas alkaline phosphatase levels were significantly increased (305+/-117 versus 337+/-118 U/l, p=0.0020). Serum ferritin levels were significantly decreased by treatment with polaprezinc (158+/-141 versus 101+/-80 ng/ml, p=0.0117). The reduction rate of ALT levels by polaprezinc was positively correlated with that of ferritin (r(2)=0.536, p=0.0389). There was a tendency toward a decrease in serum type IV collagen 7S levels after treatment with polaprezinc. However, administration of polaprezinc did not affect peripheral blood cell counts, other liver function tests, or HCV-RNA loads. CONCLUSIONS: These findings suggest that polaprezinc exerts an anti-inflammatory effect on the liver in patients with HCV-related CLD by reducing iron overload.

Successful treatment of hypovascular advanced hepatocellular carcinoma with lipiodol-targetting intervention radiology.

We report a case of hypovascular advanced hepa-tocellular carcinoma (HCC) successfully treated with a novel combination therapy of percutaneous ethanol-lipiodol injection (PELI) and intervention radiology (IVR), lipiodol-targetting IVR (Lipi-IVR). The present case had a hypovascular HCC (3 cm in diameter) located in the S6 region of the liver. Although the tumor was not detectable at all by both of early and late phase of helical dynamic computed tomography (CT), it could be detected by ultrasonography (US) as a low echoic space occupying lesion (SOL) beside the gallbladder and right kidney. Serum levels of alpha fetoprotein (AFP) and AFP-L3 were extremely high. Combination therapy of PELI, firstly reported in our department, and IVR (PELI and IVR, lipiodol-targetting IVR) was performed twice for the treatment. PELI could effectively visualize the location of the tumor for IVR treatment and show the presence of a thin blood vessel branching from the right hepatic artery flowing into the lipiodol deposit. After treatment, the serum levels of AFP and AFP-L3 were rapidly decreased to normal and maintained for more than eight months. Thus, this case expressing the tremendous effect might give us insight into the effectiveness of the novel combination therapy of PELI and IVR for the treatment of hypovascular HCC.

Blockade of renin-angiotensin system in antifibrotic therapy.

Recent studies have shown that the renin-angiotensin system (RAS) plays a pivotal role in liver fibrosis. An intrahepatic RAS is expressed in chronically damaged livers, and angiotensin-II (AT-II) reportedly stimulates contraction and proliferation of the activated hepatic stellate cells (Ac-HSC), and increases the transforming growth factor-beta (TGF-beta) expression through angiotensin type-I receptors (AT1-R). Some studies have demonstrated that the clinically used angiotensin-converting enzyme (ACE) inhibitor (ACE-I), and AT1-R blockers (ARB) significantly attenuated experimental liver fibrosis along with suppression of the Ac-HSC and hepatic TGF-beta expression. Angiotensin-II also stimulates the tissue inhibitor of metalloproteinases-1 (TIMP-1) in a dose- and time-dependent manner via protein kinase-C as an intracellular signaling cascade in the Ac-HSC, and these effects are completely suppressed by ARB. Combination treatment with low-dose interferon (IFN) and ACE-I exerts a stronger inhibitory effect than either single agent on its own. In humans it has been reported that ARB markedly improved the liver fibrosis score and TGF-beta expression in patients with chronic hepatitis C and non-alcoholic steatohepatitis. Serum fibrosis markers also significantly improved by treatment with low-dose IFN and ACE-I in patients with chronic hepatitis C, refractory to IFN monotherapy. Collectively, these data suggest that the interaction between AT-II and AT1-R plays a pivotal role in liver fibrosis development. Because both ACE-I and ARB are widely used in clinical practice without serious side-effects, these drugs in combination with IFN may provide a new strategy for antifibrosis therapy.

Utility of pancreatic duct brushing for diagnosis of pancreatic carcinoma.

BACKGROUND: The aim of this study was to evaluate the usefulness of pancreatic duct brushing for diagnosis of pancreatic carcinoma. METHODS: Brush cytology was attempted in 58 patients suspected of having pancreatic malignancy because of stricture of the main pancreatic duct, confirmed by endoscopic retrograde cholangiopancreatography. Thirty-eight patients were finally diagnosed by an operation or the clinical course as having pancreatic carcinoma, and the remaining 20 patients as having chronic pancreatitis. The usefulness of brush cytology for diagnosis of pancreatic carcinoma was estimated. We interpreted failures of pancreatic duct brushing to be false negatives when the lesion was malignant. RESULTS: In 48 of 58 patients (82.8%), brushing was successfully performed and satisfactory specimens were obtained. Brush cytology was positive in 25 of 38 patients with pancreatic carcinoma (sensitivity 65.8%) and negative in all patients without malignancy (specificity 100%). Overall accuracy was 76.4%. During 2001-2005, the number of back-and-forth motions of the brush was increased to more than 30 times. The sensitivity significantly improved from 43.8% in 1997-2000 to 81.8% in 2001-2005 (P < 0.05). The increased success rate of brushing by improvement of skill in manipulating the guidewire and increased number of cells smeared on glass slides by increased back-and-forth motion of the brush may account for this improvement over time. Moreover, the sensitivity in 2001-2005 was 85.7% if failures of brushing with pancreatic carcinoma are excluded. No major complications occurred, except for two patients with a moderate grade of acute pancreatitis. CONCLUSIONS: Although further studies with a large number of patients are needed, our results suggest that with recent improvements of the brushing technique, pancreatic duct brushing is a useful and safe method for the differential diagnosis of malignancy from benign diseases of the pancreas.

Usefulness of single and repetitive percutaneous transhepatic gallbladder aspiration for the treatment of acute cholecystitis.

BACKGROUND: The aim of this study was to evaluate the safety and usefulness of single and repetitive percutaneous transhepatic gallbladder aspiration (PTGBA) for the treatment of acute cholecystitis. METHODS: PTGBA was performed in patients with acute cholecystitis who showed no improvement after treatment with broad-spectrum antibiotics. PTGBA was carried out at bedside. When the bile was too thick to be aspirated through a 21-gauge needle, an 18-gauge needle was used. Aspiration of the gallbladder contents and injection of antibiotics into the gallbladder were performed without the placement of a drainage catheter. When improvement was not observed after the first attempt, PTGBA was repeated. RESULTS: Single PTGBA achieved improvement in 32 of 45 patients. In 11 of the remaining 13 patients, the second PTGBA was effective. In the remaining two patients, repetitive PTGBA was not carried out because of advanced cancer. In two of 45 patients, 18-gauge needles were necessary for PTGBA because of the high viscosity of the bile. PTGBA was carried out in three patients with blockage of the cystic duct by a stent, and it was effective in all three. Two patients whose conditions improved with a single PTGBA experienced a recurrence at 4 and 31 months, respectively, after PTGBA. No other severe complications related to PTGBA were observed in any patients. CONCLUSIONS: For the treatment of acute cholecystitis that does not react to conservative therapies, PTGBA is a safe, simple, and effective treatment modality that can be performed at bedside without any severe complications.

Vitamins K2, K3 and K5 exert antitumor effects on established colorectal cancer in mice by inducing apoptotic death of tumor cells.

Although a number of studies have shown that vitamin K possesses antitumor activities on various neoplastic cell lines, there are few reports demonstrating in vivo antitumor effects of vitamin K, and the antitumor effect on colorectal cancer (CRC) remains to be examined. Therefore, antitumor effects of vitamin K on CRC were examined both in vitro and in vivo. Vitamins K2, K3 and K5 suppressed the proliferation of colon 26 cells in a dose-dependent manner, while vitamin K1 did not. On flow cytometry, induction of apoptosis by vitamins K2, K3 and K5 was suggested by population in sub-G1 phase of the cell cycle. Hoechst 33342 staining and a two-color flow cytometric assay using fluorescein isothiocyanate-conjugated annexin V and propidium iodide confirmed that vitamins K2, K3 and K5 induced apoptotic death of colon 26 cells. Enzymatic activity of caspase-3 in colon 26 cells was significantly up-regulated by vitamins K2, K3 and K5. The pan-caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone, substantially prevented vitamin K-mediated apoptosis. In vivo study using syngeneic mice with subcutaneously established colon 26 tumors demonstrated that intravenous administration of vitamins K2, K3 and K5 significantly suppressed the tumor growth. The number of apoptotic tumor cells was significantly larger in the vitamin K-treated groups than in the control group. These results suggest that vitamins K2, K3 and K5 exerted effective antitumor effects on CRC in vitro and in vivo by inducing caspase-dependent apoptotic death of tumor cells, suggesting that these K vitamins may be promising agents for the treatment of patients with CRC.

[Gastroesophageal reflux disease in diabetic patients].

Esophageal dysfunctions occur frequently in patients with diabetic neuropathy, and the complication of gastroesophageal reflux disease(GERD) has also been reported. However, the characteristics of GERD complicated with diabetes mellitus are obscure because no detailed assessment was performed. We performed long-term ambulatory esophageal pH and motility monitoring in diabetic patients, and analyzed the data in detail. Esophageal dysfunctions including GERD were significantly related to diabetic neuropathy and duration. Although GERD was frequently complicated with diabetes mellitus, the symptoms were not apparent in diabetic patients. The average score of a questionnaire was extremely low, and no significant difference in score was observed between patients with and without gastroesophageal reflux. We should keep in mind that diabetic patients have the abnormal gastroesophageal reflux even when they do not complain of the related symptoms.

Abundance of microRNA target motifs in the 3'-UTRs of 20527 human genes.

A mechanism, selective avoidance, proposes that microRNA (miRNA) target sites are selectively depleted in the 3'-UTRs of genes expressed at the same time and place as a miRNA. If this mechanism is ubiquitous, the target motif occurrences in the 3'-UTRs would be decreased. To test this hypothesis, we examined miRNA target motif occurrences in the 3'- and 5'-UTRs of 20527 human protein-coding genes. The results revealed that miRNA target motifs appeared more frequently than non-target motifs and were enriched in the 3'-UTRs. This enrichment was relatively reduced in a set of 2525 genes coexpressed with miR-124a in the prefrontal cortex, but still remained at a high level, suggesting that miRNA target motifs are fostered by some other factors that surpass the influence of selective avoidance.