Piperic acid derivative as a molecular modulator to accelerate the IAPP aggregation process and alter its antimicrobial activity.

Piperic acid derivatives were found to affect the islet amyloid polypeptide (IAPP) aggregation process. Structure-activity relationship studies revealed that PAD-13 was an efficient molecular modulator to accelerate IAPP fibril formation by promoting primary and secondary nucleation and reducing its antimicrobial activity.

[Temporal and spatial variations of soil moisture in North China]. / 华北地区土壤水分的时空变化特征.

Soil moisture is an important hydrological parameter and a basic element for research in water cycle and climate change. Using hourly recorded soil moisture data of 374 stations from the new generation of automatic soil moisture encryption observation network constructed by China Meteo-rological Administration, we analyzed the spatial and temporal variations of soil moisture in North China from 2013 to 2019 and the relationship with precipitation and temperature. The results showed that soil moisture in 10-100 cm layer decreased fluctuatly as a whole, with the decreasing at the 100 cm being serious. The spatial distribution of soil moisture at different depths was characterized by high in the southeast and low in the northwest. About 63% of the surface layer was short of moisture. Soil moisture at different depths changed significantly with the seasons. In summer, soil moisture of each layer reached the highest and soil entropy of each layer was suitable, while it reached a low point in spring. Soil moisture was closely correlated with precipitation and temperature, but the correlation got weaker gradually with the increase of soil depth. Soil moisture was more sensitive to precipitation than to temperature.

Advances with Long Non-Coding RNAs in Diabetic Peripheral Neuropathy.

Long non-coding RNAs (lncRNAs) are a group of non-coding RNAs longer than 200 nucleotides, which are defined as transcripts. The lncRNAs are involved in regulating gene expression at epigenetic, transcriptional, and post-transcriptional levels. Recent studies have found that lncRNA is closely related to many diseases like neurological diseases, endocrine and metabolic disorders. Diabetic peripheral neuropathy (DPN) is one of the most common chronic complications of diabetes mellitus. In this review, we highlight the latest research related to lncRNAs in DPN.

Long Non­Coding RNAs Regulate Inflammation in Diabetic Peripheral Neuropathy by Acting as ceRNAs Targeting miR-146a-5p.

BACKGROUND: Long non-coding RNAs (lncRNAs), as competing endogenous RNAs (ceRNAs), can regulate various pathophysiological processes by binding competitively to microRNAs at the post-transcription level. Our previous work demonstrated that miR-146a-5p was lowly expressed in diabetic peripheral neuropathy (DPN) rats. However, the ceRNA network in DPN mediated by lncRNAs and miR-146a-5p remains to be explored. METHODS: Two groups of rats (n=4 per group), a type 2 diabetes (T2DM) group and a DPN group, were used in this study. Sciatic nerve conduction velocity (NCV) of each rat was determined at the 6th and the 12th week. LncRNA microarray analysis was performed in the sciatic nerve of DPN and T2DM rats. Based on the TargetScan algorithm and the miRanda database, we determined the differentially expressed (DE) lncRNAs bound to miR-146a-5p. Furthermore, we verified the DE lncRNAs potentially bound to miR-146a-5p by qRT-PCR. The genes targeted by miR-146a-5p were identified by bioinformatics prediction and experimental techniques. RESULTS: We found 413 DE lncRNAs between DPN and T2DM rats (|log2FC| ≥ 2 and adjust P ≤ 0.05). Eight DE lncRNAs were predicted to bind to miR-146a-5p by both algorithms, of which four were verified by qRT-PCR. TRAF6, IRAK1, and SMAD4 were identified as miR-146a-5p targeted genes and were predominantly enriched in the inflammatory signaling pathway. CONCLUSION: LncRNAs may contribute to the pathogenesis of DPN by regulating inflammation through functioning as ceRNAs of miR-146a-5p.

MicroRNA-146a-5p Downregulates the Expression of P-Glycoprotein in Rats with Lithium-Pilocarpine-Induced Status Epilepticus.

Increasing evidence supports that the efflux transporters, especially P-glycoprotein (P-gp), have vital roles on drug resistance in epilepsy. Overexpression of P-gp in the brain could reduce the anti-epileptic drugs (AEDs) concentration in the epileptogenic zone, resulting in drug resistance. Studies have demonstrated that recurrent seizures induce the expression of P-gp and status epilepticus (SE) could upregulate the expression of P-gp, resulting in drug resistance. MicroRNAs (miRNAs), as endogenous regulators, represent small regulatory RNA molecules that have been shown to act as negative regulators of gene expression in different biological processes. We investigated the impact of miR-146a-5p on the expression of P-gp in status epilepticus rat model. The expression of miR-146a-5p in rat cortex and hippocampus was measured by quantitative RT-PCR at 2 weeks after induction of SE. Meanwhile, we detected the expression of P-gp in the brain of SE rats using Western blotting and immunohistochemistry. Upregulation of miR-146a-5p and overexpression of P-gp were evident at 2 weeks after SE. Moreover, the expression of P-gp was downregulated by injection of miR-146a mimic into the hippocampus. We also detected the expression of interleukin-1 receptor-associated protein kinases-1 (IRAK1) and tumor necrosis factor receptor-associated factor 6 (TRAF6) and nuclear factor-kappaB (NF-κB) p65 using Western blotting and immunohistochemistry, which indicated the expression of IRAK1, TRAF6 and NF-κB p-p65/p65 increased in the brain of SE rats, and overexpression of miR-146a-5p could downregulate the expression of IRAK1, TRAF6, NF-κB p-p65/p65 and P-gp. Our study indicated that miR-146a-5p may decrease the expression of P-gp in status epilepticus rats via NF-κB signaling pathway.

The Neuroprotective Effect of Astaxanthin on Pilocarpine-Induced Status Epilepticus in Rats.

Cognitive dysfunction is one of the serious complications induced by status epilepticus (SE), which has a significant negative impact on patients' quality of life. Previous studies demonstrated that the pathophysiological changes after SE such as oxidative stress, inflammatory reaction contribute to neuronal damage. A recent study indicated that preventive astaxanthin (AST) alleviated epilepsy-induced oxidative stress and neuronal apoptosis in the brain. In the present study, rats were treated with vehicle or AST 1 h after SE onset and were injected once every other day for 2 weeks (total of seven times). The results showed that the cognitive function in SE rats was significantly impaired, and AST treatment improved cognitive function in the Morris water maze (MWM). Magnetic resonance imaging (MRI), hematoxylin-eosin (HE) staining and TdT-mediated dUTP Nick-End Labeling (TUNEL) staining showed obvious damage in the hippocampus of SE rats, and AST alleviated the damage. Subsequently, we evaluated the effect of AST on relative pathophysiology to elucidate the possible mechanisms. To evaluate the oxidative stress, the expression of malondialdehyde (MDA) and superoxide dismutase (SOD) in plasma were detected using commercially available kits. NADPH oxidase-4 (Nox-4), p22phox, NF-E2-related factor 2 (Nrf-2), heme oxygenase 1 (Ho-1) and sod1 in the parahippocampal cortex and hippocampus were detected using western blot and real-time polymerase chain reaction (RT-PCR). The levels of MDA in plasma and Nox-4 and p22phox in the brain increased in SE rats, and the levels of SOD in plasma and Nrf-2, Ho-1 and sod1 in the brain decreased. Treatment with AST alleviated these changes. We also detected the levels of inflammatory mediators like cyclooxygenase-2 (cox-2), interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α) and NF-κB phosphorylation p65 (p-p65)/p65 in the brain. The inflammatory reaction was significantly activated in the brain of SE rats, and AST alleviated neuroinflammation. We detected the levels of p-Akt, Akt, B-cell lymphoma-2 (Bcl-2), Bax, cleaved caspase-3, and caspase-3 in the parahippocampal cortex and hippocampus using western blot. The levels of p-Akt/Akt and Bcl-2 decreased in SE rats, Bax and cleaved caspase-3/caspase-3 increased, while AST alleviated these changes. The present study indicated that AST exerted an reobvious neuroprotective effect in pilocarpine-induced SE rats.

Nanoparticle-microRNA-146a-5p polyplexes ameliorate diabetic peripheral neuropathy by modulating inflammation and apoptosis.

Nontoxic and nonimmunogenic nanoparticles play an increasingly important role in the application of pharmaceutical nanocarriers. The pathogenesis of diabetic peripheral neuropathy (DPN) has been extensively studied. However, the role of microRNAs in DPN remains to be clarified. We verified in vitro that miR-146a-5p mimics inhibited the expression of proinflammatory cytokines and apoptosis. Then, we explored the protective effect of nanoparticle-miRNA-146a-5p polyplexes (nano-miR-146a-5p) on DPN rats. We demonstrated that nano-miR-146a-5p improved nerve conduction velocity and alleviated the morphological damage and demyelination of the sciatic nerve of DPN rats. The expression of the inflammatory cytokines, caspase-3, and cleaved caspase-3 in the sciatic nerve was inhibited by nano-miR-146a-5p. Additionally, nano-miR-146a-5p increased the expression of myelin basic protein. These results all indicated that nano-miR-146a-5p had a protective effect on peripheral nerves in the DPN rat model, which may occur through the regulation of the inflammatory response and apoptosis.

The Protective Effect of Astaxanthin on Cognitive Function via Inhibition of Oxidative Stress and Inflammation in the Brains of Chronic T2DM Rats.

Currently, there are no effective treatments for diabetes-related cognitive dysfunction. Astaxanthin (AST), the most powerful antioxidant in nature, exhibits diverse biological functions. In this study, we tried to explore whether AST would ameliorate cognitive dysfunction in chronic type 2 diabetes mellitus (T2DM) rats. The T2DM rat model was induced via intraperitoneal injection of streptozotocin. Forty Wistar rats were divided into a normal control group, an acute T2DM group, a chronic T2DM group, and an AST group (treated with AST at a dose of 25 mg/kg three times a week). The Morris water maze test showed that the percentage of time spent in the target quadrant of the AST group was identical to that of the chronic T2DM group, while the escape latency of the AST group was decreased in comparison to that of the chronic T2DM group. Histology of the hippocampus revealed that AST ameliorated the impairment in the neurons of diabetic rats. Western blot showed that AST could upregulate nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) expression and inhibit nuclear transcription factor kappa B (NF-κB) p65 activation in the hippocampus. We found that AST increased the level of superoxide dismutase (SOD) and decreased the level of malondialdehyde (MDA) in the hippocampus. In addition, the levels of interleukin 1 beta (IL-1ß) and interleukin 6 (IL-6) were reduced in the AST group compared with those in the chronic T2DM group. The findings of this research imply that AST might inhibit oxidative stress and inflammatory responses by activating the Nrf2-ARE signaling pathway.

MicroRNA-146a: A Comprehensive Indicator of Inflammation and Oxidative Stress Status Induced in the Brain of Chronic T2DM Rats.

Objective: It was demonstrated that inflammation and oxidative stress induced by hyperglycemia were closely associated with alteration of miR-146a. Here, we investigated the role of miR-146a in mediating inflammation and oxidative stress in the brain of chronic T2DM rats. Methods: The chronic T2DM (cT2DM) models were induced by intraperitoneal administration of STZ (35 mg/kg) after being fed a high-fat, high-sugar diet for 6 weeks. H&E staining was conducted to observe the morphological impairment of the rat hippocampus. The expressions of inflammatory mediators (COX-2, TNF-α, IL-1ß) and antioxidant proteins (Nrf2, HO-1) were measured by western blot. The levels of MDA and SOD were detected by the respective activity assay kit. The levels of p22phox and miR-146a were examined by quantitative real-time PCR (qRT-PCR). The expressions of IRAK1, TRAF6 and NF-κB p65 were measured by western blot and qRT-PCR. Pearson correlation analysis was performed to investigate the correlations between miR-146a and inflammatory mediators as well as oxidative stress indicators. Results: The expression of miR-146a was negatively correlated with inflammation and oxidative stress status. In the brain tissues of cT2DM rats, it was observed that the expressions of inflammatory mediators (COX-2, TNF-α, IL-1ß) and oxidative stress indicators including MDA and p22phox were elevated, which were negatively correlated with the expression of miR-146a. While, the antioxidant proteins (Nrf2, HO-1, SOD) levels decreased in the brain of cT2DM rats, which were positively correlated with the miR-146a level. The expressions of NF-κB p65 and its specific modulators (IRAK1&TRAF6) were elevated in the brain of cT2DM rats, which might be inhibited by miR-146a. Conclusion: Our results implied that increased inflammation and oxidative stress status were associated with brain impairment in cT2DM rats, which were negatively correlated with miR-146a expression. Thus, miR-146a may serve as a negative comprehensive indicator of inflammation and oxidative stress status in the brain of chronic T2DM rats.

Involvement of microRNA-146a in diabetic peripheral neuropathy through the regulation of inflammation.

PURPOSE: Recent evidence has shown the involvement of inflammation in the development of diabetic peripheral neuropathy (DPN). MicroRNA-146a (miR-146a) is closely involved in the inflammatory response. However, the role of miR-146a in the inflammatory reaction in DPN has not been clarified. This study was designed to explore the role of miR-146a in the regulation of inflammatory responses in DPN. METHODS: Rats were randomly divided into three groups (n=6 per group): control group, type 2 diabetes mellitus (T2DM) group and DPN group. T2DM and DPN rats were intraperitoneally injected with streptozotocin. Sciatic nerve conduction velocity (NCV) was determined at the 6th week and the 12th week in each group. The expression of microRNAs was detected by quantitative real-time polymerase chain reaction in three sciatic nerves for each group of rats. Expression of inflammatory cytokines in nerve tissues and plasma was measured by Western blot and Bio-Plex Pro™ assays. RESULTS: The NCV and expression levels of miR-146a in the DPN group were significantly decreased (P<0.01) compared to the other two groups. Expression of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1ß) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in the DPN group was significantly increased compared with the control and T2DM groups (P<0.01). Pearson's correlation analysis showed that the expression level of miR-146a was negatively correlated with the levels of IL-1ß, TNF-α and NF-κB. CONCLUSION: miR-146a is involved in the pathogenesis of DPN, and its expression level is closely related to the inflammatory responses that aggravate sciatic nerve injuries.

Aliskiren add-on therapy effectively reduces proteinuria in chronic kidney disease: an open-label prospective trial.

INTRODUCTION: The combination therapy of aliskiren and renin-angiotensin-aldosterone system (RAAS) blocker in chronic kidney disease (CKD) is controversial. Whether such dual blockade can effectively apply to patients with CKD irrespective of stage and amount of proteinuria remains uncertain. METHODS: We added aliskiren at a dosage of 150 mg/day for six months in 103 Chinese CKD patients who had been treated with angiotensin converting enzyme inhibitors (ACEis) or angiotensin receptor blockers (ARBs) and still had significant proteinuria or uncontrolled hypertension. Blood pressure, serum creatinine, estimated glomerular filtration rate (eGFR), potassium, and spot urine protein-to-creatinine ratio (UPCR) were measured at three and six months after aliskiren add-on therapy and compared with baseline. RESULTS: The combination of aliskiren and ACEi or ARB significantly reduced UPCR by 23% (p=0.001) and mean arterial pressure by 7.9 ± 13.8 mmHg (p<0.001) at six months. Twenty-five percent of subjects had a greater than 50% reduction in UPCR. No significant changes in eGFR and serum potassium level were noted at six months. CONCLUSIONS: Adding aliskiren on ACEi or ARB in CKD patients, both in diabetes and non-diabetes, has a favorable effect on reducing residual proteinuria and inadequately controlled blood pressure.

Ultrasound localization of the femoral vein facilitates successful cannulation for hemodialysis.

BACKGROUND: Real-time ultrasound-guided techniques allow for improved cannulation of the internal jugular vein and femoral vein for hemodialysis; however, these techniques require extra sterilization procedures, specialized probes, or needle guides. A simpler ultrasound vessel localization method was performed to investigate whether this alternative approach would aid in the cannulation of the femoral vein for patients in whom temporary angioaccess was required for hemodialysis. METHODS: Patients requiring temporary femoral vein catheters for hemodialysis were divided into 2 groups on alternating days of the week during a 6-month period. One group underwent ultrasound localization of the femoral vein before cannulation and the second group received conventional landmark localization. Data regarding the strength of the femoral arterial pulse, number of attempts, failures, and complications were recorded. RESULTS: Ultrasound localization resulted in significantly improved first-attempt success rates, reduced attempts, and reduced failure and complication rates overall (p < 0.001, p < 0.001, p = 0.002 and p = 0.004 respectively) as well as in the group of patients with a clearly discernible arterial pulse (p < 0.001, p = 0.001, p = 0.004 and p = 0.011 respectively). The same trend was observed among patients with faintly palpable or non-palpable femoral arterial pulses, although the differences were not statistically significant. CONCLUSION: Cannulation of the femoral vein for hemodialysis should be performed with the aid of ultrasound. If real-time ultrasound-guided cannulation is not available, the vessel localization method is a good alternative, given its known limitations and the fact that it is simpler. It remains to be determined whether 1 - dimensional localization or localization including vessel depth information can improve outcomes in patients with faintly palpable or non-palpable femoral arterial pulses.

Free p-cresol sulfate is associated with survival and function of vascular access in chronic hemodialysis patients.

BACKGROUND: Protein-bound uremic toxins indoxyl sulfate (IS) and p-cresol sulfate (p-CS) have been implicated as an important factor in uremic syndrome. Recent evidence indicates that both IS and p-CS are predictors of cardiovascular as well as all-cause mortality among chronic dialysis patients. We conducted a study to analyze the relationship between IS and p-CS and vascular access (VA) outcome in chronic hemodialysis (HD) patients. METHODS: A total of 91 chronic stable HD patients were divided into groups according to survival of VA and frequency of VA dysfunction. Demographic and biochemical data were reviewed and recorded. Serum levels of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1, and the total and free forms of IS and p-CS were determined. RESULTS: Patients with a history of frequent VA failure and dysfunction had lower albumin and higher levels of ICAM-1, free IS, free and total p-CS. Diabetes was associated with higher IS and p-CS. Logistic regression revealed that diabetes and free p-CS were independent factors associated with poor outcome of VA. CONCLUSION: Endothelial dysfunction and uremic toxins were associated with survival and function of VA. Diabetes and free p-CS were significantly related to the outcome of VA among chronic HD patients.

Albuminuria, proteinuria, and urinary albumin to protein ratio in chronic kidney disease.

BACKGROUND: Both albuminuria and proteinuria are important disease markers of chronic kidney disease (CKD). Their relationship and the ratio between urinary albumin and protein in patients with CKD have not been investigated. Whether clinical features can affect these measurements is not clear. METHODS: We conducted a cross-sectional study in 602 CKD patients. Demographic data, including age, gender, and co-morbidity such as diabetes, hypertension, hyperuricemia, and hyperlipidemia, were reviewed and recorded. Their urinary albumin, total protein, and creatinine were determined and urinary albumin to creatinine ratio (UACR), total protein to creatinine ratio (UPCR), and albumin to total protein ratio (UAPR) were calculated. Their estimated glomerular filtration rate (eGFR) was calculated according to serum creatinine. The correlation between UACR and UPCR was thus analyzed. We also investigated factors associated with these urinary measurements. RESULTS: UACR and UPCR increased progressively as renal function deteriorated, while UAPR increased to a plateau in CKD stage 4. There was direct relationship between UACR and UPCR. UAPR rose exponentially with the increase of both UACR and UPCR when UACR <500 mg/g or UPCR <1,000 mg/g. Multivariate regression analysis revealed diabetes and hyperuricemia were associated with increased UACR and UPCR, while both urinary parameters were inversely related to male gender and eGFR. Diabetes and hyperuricemia were associated with increased UAPR and UAPR was negatively correlated with age and eGFR. CONCLUSION: There was a significant association between UACR and UPCR in patients with CKD. Characteristics of patients, renal function, and co-morbidities all affected UACR, UPCR, and UAPR.

Development of cephradine-induced acquired factor V inhibitors: a case report.

OBJECTIVE: To report a case of cephalosporin-induced factor V inhibitor development, an uncommon but potentially fatal condition characterized by severe hemorrhage. CASE SUMMARY: A 71-year-old Chinese man presented with factor V inhibitors after a 7-day cephradine course for a urinary tract infection, characterized by abnormal prothrombin time (PT) and activated partial thromboplastin time (aPTT), gross hematuria, upper gastrointestinal bleeding, and left groin hematoma. Systemic corticosteroid administration restored his factor V activity levels, PT, and aPTT to within normal limits, and hemorrhagic symptoms resolved. Three weeks after successful treatment of bleeding diathesis, he received another 8-day cephradine course for cellulitis. After another 4 weeks, he suffered from recurrent factor V inhibitors presented with abnormal PT, aPTT, and upper gastrointestinal bleeding. The patient eventually died due to hemorrhagic shock despite a second course of corticosteroids. DISCUSSION: Cephalosporins are known to cause coagulopathy via hypoprothrombinemia. Another pathway seldom mentioned in the literature is factor V inhibitor induction, which may result in factor V deficiency. In our patient, factor V deficiency due to inhibitors developed each time that the patient received repeated cephradine treatment. According to the Naranjo probability scale, the relation between the formation of factor V inhibitors and cephradine treatment was probable. CONCLUSIONS: Because cephalosporins are commonly used for their wide therapeutic index and few adverse effects, iatrogenic complications associated with these drugs may be neglected or underdiagnosed. On the basis of our patient's report, careful review of medical records to avoid reexposure to the offending drug cannot be overemphasized.

A classification of NOergic neurons in the inferior colliculus of rat according to co-existence with classical amino acid transmitters.

Since the localization of nitric oxide synthase (NOS) can be identified by enzyme histochemistry for NADPH-diaphorse (NADPH-d), this method has been used widely for mapping NOS-containing (presumably NOergic) neurons in the central nervous system. So far several studies suggest that NADPH-d is present in distinct neuronal populations in the inferior colliculus (IC), a major processing center for both the ascending and descending auditory pathway, and NO may play an important role in audition. On one hand, there is evidence from several lines of research that the IC makes extensive use of the neuroactive amino acids, in particular the inhibitory transmitter g-aminobutyric acid (GABA) and the excitatory amino acid glutamate (GLU). However, lacking is a description of the distribution of NOergic neurons to which traditional neurotransmitters may be linked. The present research utilized NADPH-d enzyme histochemistry in combination with immunocytochemistry to determine if NO may colocalize with either or both GABA and glutamate in distinct subpopulations of IC neurons. The NADPH-d positive neurons were predominantly found in two main subdivisions of the IC: the external cortex (ECIC) and the dorsal cortex (DCIC). The large numbers of these NADPH-d positive neurons appeared immunostained for GLU while only a small number, seemed to belong to the small cells (somatic area < 100 microm2) similarity to stellate cells group was positive for GABA throughout the cortex of the IC. Owing to no coexistence between GABA and GLU in the same NADPH-d positive neuron in the pairs of adjacent sections of the IC by the mirror-image technique, the present results consequently support that NOergic neurons could be subdivided into at least three distinct populations with a large proportion of about 77% being GLUergic, much lower frequency of about 11% being GABAergic and the remaining 12% expressing non-GABA and non-GLU. In summary, the existence of two functionally distinct populations of NO/GABAergic and NO/GLUergic neurons in the NOergic neurons of IC suggest that at least two differential pattern of GLU-mediated excitatory NO transmission and GABA-mediated inhibitory NO transmission are involved in the networking of auditory communication in the cortex of IC.

["Restricted ablation" of elongated uvula mucosa by the injection of the ethanol/steroid mixture: a new treatment for snoring and OSAS].

We developed a "Submucosal Ethanol/Steroid (E/S) Injection Method (SEIM)" using an injection prepared by dissolving steroid with powerful antiinflammatory effect, which has the excellent effect of contractile reduction in oral tissues. In this clinical trial, the ablation effect of SEIM on the abnormally elongated uvula and the soft palate was examined in each one clinical case of obstructive sleep apnea syndrome (OSAS) and simple snoring. In the OSAS, we found that the uvula was reduced from 15 to 10mm, the visual analog scale (VAS) of snoring was reduced from 10 to 4 points, and the respiration disturbance index of the apnea-hypopnea index (AHI) improved from 35.3 to 26.1 after treatment. In simple snoring, the uvula was reduced from 11 to 8.5mm and VAS was relieved from 7 to 2 points after treatment. Our approach will produce a great clinical significance for not only OSAS or simple snoring but also treatment of the allergic rhinitis, etc, because the contractile tissue reduction can be attained safely in these diseases without open surgical wounds and unnecessary deformation or destruction of the mucosal structure.

[Histological study on the effects of ethanol injection on mouth mucosa contraction loss].

We previously reported that a loss of contraction in the mucosal tissue of the palate arch is effectively induced by ethanol injections of moderate concentration and dosage. The present study was performed to obtain more information on how such ethanol injections induce contraction loss in mucosa tissue. Guinea pigs of both sexes were used in this study. The left arch of the palate mucosa was injected with 2 microliters of 70% ethanol and used as the experimental group. The right arch of the palate mucosa of the same animal was injected with saline and used as a control. One, three, five, eight, 10, 30, 50, and 90 days after injection, the mucosal tissues that received the injection were resected under anesthesia and processed for light microscopy using standard procedures. One day after the ethanol injection, severe coagulative degeneration of the mucosal tissue of the palate was seen. However, the damaged area was strictly restricted to the arch of the palate. Coagulative degeneration of the tissue peaked three days after the injection. Thereafter, the mucosal epithelial and mucosal connective tissues regenerated, and the damaged mucosal tissue quickly began to repair. An apparent cicatricial contraction loss was observed 10 days after the ethanol injection, along with the progression of fibrotic changes in the submucosal connective tissue of the arch of the palate. The regenerative action of the mucosal arch of the palate abated 30 days after the ethanol injection, and the reduced mucosal tissue appeared to have become denser as a result of an increase in dense fibrous connective tissue in the submucosal layer. No cell malignancies were seen throughout the entire 90-day observation period. In conclusion, cicatricial contraction loss of the mucosal arch of the palate resulting from the injection of an appropriate concentration and dosage of ethanol leads to the regeneration of the mucosal epithelium and fibrotic changes in the submucosa. The ethanol injection described here seems to be extremely safe, since it exerted no malignant effects on the cells and tissues either morphologically or functionally.

[Experimental study of contraction loss effects of mouth mucosa by ethanol injection].

We began treating patients with simple snoring and obstructive sleep apnea syndrome (OSAS) with the coblator radiofrequency generator in our outpatient clinic from April 2001. Good clinical results have been obtained, but we noticed a contractile effect on mucosa from ethanol, which possesses marked sclerotic degenerative action on tissue as well as radiofrequency energy. We conducted a series of experiments in a guinea pig model to investigate the efficacy of local ethanol injection in contracting mouth mucosa. To examine the influence on respiration of liquid injection, physiological saline was gradually injected in decrements into the arch of the palate mucosa. We found that the safe dosage that did not bring about edema and subsequent dyspnea was under 10 microliters. Based on this finding, ethanol in concentrations of 50%, 70%, and 100% at volumes of 1, 2, 4, 8, and 10 microliters was injected into the arch of the palate mucosa in guinea pigs and changes in local field mucosa were observed daily. In the 50% ethanol injection, no clear contractile effect on mucosa could be observed at any dosage. In contrast, the 100% ethanol injection led to strong tissue impairment that caused extensive necrotic collapse of the local field mucosa, even when the dosage was down to the minimum of 1 microliter. We found that, injection of 70% ethanol at 1 or 2 microliters, however, resulted in formation of a local field mucosa wound of lesser degree that healed completely within a few days, associated with moderate contraction of mucosal tissue. We concluded that in moderate dosage, 70% ethanol seems to have the potential for the treatment of endermosis, such as uvuloptosia (elongated uvula) and hypertrophy of palate mucosa, as a useful mucosa contractile agent.

[Immunohistochemical study for monoamine neurons in the brain of unilateral inner ear impaired rats].

Patients with inner ear impairment have complaints of vertigo and also occasionally depression. The present study was undertaken in order to evaluate changes in monoamines which have reportedly been closely related to depression, using cisplatin-induced unilateral inner-ear impaired rats. A dose of 0.5 mg/kg of cisplatin was injected into the right tympanic cavity under pentobarbital Na+ anesthesia. One or two weeks later, animals were fixed with paraformaldehyde, and thereafter immunohistochemical stainings for monoamine-containing cells in the brain were carried out. To visualize 5-hydroxytryptamine (5-HT), noradrenaline (NA) and dopamine (DA) neurons, we used mouse antibodies against 5-HT, NA, and DA syntheses, i.e., tryptophan hydroxylase (TRH), tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH). The number of TRH immunoreactive neurons significantly decreased in the lateral dorsal raphe nucleus of the ipsilateral side when compared with the contralateral side. The number of DA neurons, which were immunoreactive to TH, but not to DBH, significantly decreased in the hypothalamus of the ipsilateral side. The number of NA neurons which were immunoreactive to both TH and DBH significantly decreased in the locus coeruleus and ventral lateral pons of the ipsilateral side. An additional control study with saline-injected rats showed a lack of differences in monoamine syntheses between the injected and contralateral sides, the expressions of the synthesis on both sides being similar to that obtained in the contralateral side in cisplatin-injected rats. These results indicated the decreases in monoamine syntheses at the ipsilateral side only in the cisplatin-administered rats. We conclude that inner ear impairment may diminish the ipsilateral amount of monoamines in the brain but not the cotralateral, possibly inducing a vestibular compensation such as an upregulation of monoamine receptors.