Duodenal-jejunal bypass improves hypothalamic oxidative stress and inflammation in diabetic rats via glucagon-like peptide 1-mediated Nrf2/HO-1 signaling.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is often accompanied by impaired glucose utilization in the brain, leading to oxidative stress, neuronal cell injury and infla-mmation. Previous studies have shown that duodenal jejunal bypass (DJB) surgery significantly improves brain glucose metabolism in T2DM rats, the role and the metabolism of DJB in improving brain oxidative stress and inflammation condition in T2DM rats remain unclear. AIM: To investigate the role and metabolism of DJB in improving hypothalamic oxidative stress and inflammation condition in T2DM rats. METHODS: A T2DM rat model was induced via a high-glucose and high-fat diet, combined with a low-dose streptozotocin injection. T2DM rats were divided into DJB operation and Sham operation groups. DJB surgical intervention was carried out on T2DM rats. The differential expression of hypothalamic proteins was analyzed using quantitative proteomics analysis. Proteins related to oxidative stress, inflammation, and neuronal injury in the hypothalamus of T2DM rats were analyzed by flow cytometry, quantitative real-time PCR, Western blotting, and immunofluorescence. RESULTS: Quantitative proteomics analysis showed significant differences in proteins related to oxidative stress, inflammation, and neuronal injury in the hypothalamus of rats with T2DM-DJB after DJB surgery, compared to the T2DM-Sham groups of rats. Oxidative stress-related proteins (glucagon-like peptide 1 receptor, Nrf2, and HO-1) were significantly increased (P < 0.05) in the hypothalamus of rats with T2DM after DJB surgery. DJB surgery significantly reduced (P < 0.05) hypothalamic inflammation in T2DM rats by inhibiting the activation of NF-κB and decreasing the expression of interleukin (IL)-1ß and IL-6. DJB surgery significantly reduced (P < 0.05) the expression of factors related to neuronal injury (glial fibrillary acidic protein and Caspase-3) in the hypothalamus of T2DM rats and upregulated (P < 0.05) the expression of neuroprotective factors (C-fos, Ki67, Bcl-2, and BDNF), thereby reducing hypothalamic injury in T2DM rats. CONCLUSION: DJB surgery improve oxidative stress and inflammation in the hypothalamus of T2DM rats and reduce neuronal cell injury by activating the glucagon-like peptide 1 receptor-mediated Nrf2/HO-1 signaling pathway.

Dietary steroids promote body weight growth and induce gametogenesis by increasing the expressions of genes related to cell proliferation of sea cucumber (Apostichopus japonicus).

Steroids play a vital role in animal survival, promoting growth and development when administered appropriate concentration exogenously. However, it remains unclear whether steroids can induce gonadal development and the underlying mechanism. This study assessed sea cucumber weights post-culturing, employing paraffin sections and RNA sequencing (RNA-seq) to explore gonadal changes and gene expression in response to exogenous steroid addition. Testosterone and cholesterol, dissolved in absolute ethanol, were incorporated into sea cucumber diets. After 30 days, testosterone and cholesterol significantly increased sea cucumber weights, with the total weight of experimental groups surpassing the control. The testosterone-fed group exhibited significantly higher eviscerated weight than the control group. In addition, dietary steroids influenced gonad morphology and upregulated genes related to cell proliferation，such as RPL35, PC, eLF-1, MPC2, ADCY10 and CYP2C18. Thees upregulated differentially expressed genes were significantly enriched in the organic system, metabolism, genetic information and environmental information categories. These findings imply that steroids may contribute to the growth and the process of genetic information translation and protein synthesis essential for gonadal development and gametogenesis.

Ventromedial Thalamus-Projecting DCN Neurons Modulate Associative Sensorimotor Responses in Mice.

The deep cerebellar nuclei (DCN) integrate various inputs to the cerebellum and form the final cerebellar outputs critical for associative sensorimotor learning. However, the functional relevance of distinct neuronal subpopulations within the DCN remains poorly understood. Here, we examined a subpopulation of mouse DCN neurons whose axons specifically project to the ventromedial (Vm) thalamus (DCNVm neurons), and found that these neurons represent a specific subset of DCN units whose activity varies with trace eyeblink conditioning (tEBC), a classical associative sensorimotor learning task. Upon conditioning, the activity of DCNVm neurons signaled the performance of conditioned eyeblink responses (CRs). Optogenetic activation and inhibition of the DCNVm neurons in well-trained mice amplified and diminished the CRs, respectively. Chemogenetic manipulation of the DCNVm neurons had no effects on non-associative motor coordination. Furthermore, optogenetic activation of the DCNVm neurons caused rapid elevated firing activity in the cingulate cortex, a brain area critical for bridging the time gap between sensory stimuli and motor execution during tEBC. Together, our data highlights DCNVm neurons' function and delineates their kinematic parameters that modulate the strength of associative sensorimotor responses.

Identification of DNA Repair-Related Genes Predicting Clinical Outcome for Thyroid Cancer.

Recent studies have demonstrated the utility and superiority of DNA repair-related genes as novel biomarkers for cancer diagnosis, prognosis, and therapy. Here, we aimed to screen the potential survival-related DNA repair-related genes in thyroid cancer (TC). TCGA datasets were utilized to analyze the differentially expressed DNA repair-related genes between TC and nontumor tissues. The K-M approach and univariate analysis were employed to screen survival-related genes. RT-PCR was employed to examine the expression of DNA repair-related genes in TC samples and matched noncancer samples. CCK-8 analyses were used to determine cellular proliferation. Herein, our team discovered that the expression of four DNA repair-related genes was remarkably upregulated in TC samples in contrast to noncancer samples. Survival assays identified 14 DNA repair-related genes. In our cohort, we observed that the expression of TAF13 and DCTN4 was distinctly elevated in TC specimens in contrast to nontumor specimens. Moreover, knockdown of TAF13 and DCTN4 was observed to inhibit the TC cellular proliferation. Overall, the upregulation of TAF13 and DCTN4 is related to decreased overall survival in TC patients. Therefore, the assessment of TAF13 and DCTN4 expression may be useful for predicting prognosis in these patients.

Enhancement of Copper Uptake of Yeast Through Systematic Optimization of Medium and the Cultivation Process of Saccharomyces cerevisiae.

Copper is an essential trace element for living organisms. Copper enriched by yeast of Saccharomyces cerevisiae is regarded as the biologically available organic copper supplement with great potentiality for application. However, the lower uptake ratio of copper ions makes the production of copper enriched by yeast uneconomically and environmentally unfriendly. In this study, S. cerevisiae Cu-5 with higher copper tolerance and intracellular copper accumulation was obtained by screening of our yeast strains collection. To increase the uptake ratio of copper ions, the medium composition and cultivation conditions for strain Cu-5 were optimized systematically. A medium comprised of glucose, yeast extract, (NH4)2SO4, and inorganic salts was determined, then a novel cultivation strategy including pH control at 5.5 and increasing amounts of yeast extract for a higher concentration of copper ion in the medium was developed. The uptake ratios of copper ions were more than 90% after combining 50 to 100 mg/L copper ions with 3.5 to 5.0 g/L yeast extract, which is the highest until now and is conducive to the cost-effective and environmentally friendly production of bioactive copper in yeast-enriched form.

Neuronal Activity in the Cerebellum During the Sleep-Wakefulness Transition in Mice.

Cerebellar malfunction can lead to sleep disturbance such as excessive daytime sleepiness, suggesting that the cerebellum may be involved in regulating sleep and/or wakefulness. However, understanding the features of cerebellar regulation in sleep and wakefulness states requires a detailed characterization of neuronal activity within this area. By performing multiple-unit recordings in mice, we showed that Purkinje cells (PCs) in the cerebellar cortex exhibited increased firing activity prior to the transition from sleep to wakefulness. Notably, the increased PC activity resulted from the inputs of low-frequency non-PC units in the cerebellar cortex. Moreover, the increased PC activity was accompanied by decreased activity in neurons of the deep cerebellar nuclei at the non-rapid eye-movement sleep-wakefulness transition. Our results provide in vivo electrophysiological evidence that the cerebellum has the potential to actively regulate the sleep-wakefulness transition.

A method for combining multiple-units readout of optogenetic control with natural stimulation-evoked eyeblink conditioning in freely-moving mice.

A growing pool of transgenic mice expressing Cre-recombinases, together with Cre-dependent opsin viruses, provide good tools to manipulate specific neural circuits related to eyeblink conditioning (EBC). However, currently available methods do not enable to get fast and precise readout of optogenetic control when the freely-moving mice are receiving EBC training. In the current study, we describe a laser diode (LD)-optical fiber (OF)-Tetrode assembly that allows for simultaneous multiple units recording and optical stimulation. Since the numbers of various cables that require to be connected are minimized, the LD-OF-Tetrode assembly can be combined with CS-US delivery apparatus for revealing the effects of optical stimulation on EBC in freely- moving mice. Moreover, this combination of techniques can be utilized to optogenetically intervene in hippocampal neuronal activities during the post-conditioning sleep in a closed-loop manner. This novel device thus enhances our ability to explore how specific neuronal assembly contributes to associative motor memory in vivo.

Spontaneous recovery of conditioned eyeblink responses is associated with transiently decreased cerebellar theta activity in guinea pigs.

Behavioral studies have demonstrated that extinguished conditioned eyeblink responses (CR) can spontaneously recover after extinction. However, the neural mechanisms underlying this process are still unclear. We have shown that spontaneous cerebellar theta activity was predictive of subsequent CR extinction. Here, we sought to further evaluate the association between spontaneous recovery and cerebellar theta activity in behaving guinea pigs. It was found that trace conditioning training significantly diminished the degree of spontaneous recovery during extinction sessions as compared to delay training. Moreover, by recording local field potential in the cerebellum of guinea pigs undergoing an eyeblink conditioning extinction task, we found that spontaneous recovery of delay-paradigm CRs was associated with transiently decreased CS-evoked theta activity in the cerebellum. These findings suggest that decreased CS-evoked cerebellar theta activity may contribute to the neural process that is important for the spontaneous recovery of extinguished motor memory. Future studies are needed to clarify the neural mechanism underlying changed cerebellar theta activity during altered behavioral contingencies.

Nanochannels Photoelectrochemical Biosensor.

Nanochannels have brought new opportunities for biosensor development. Herein, we present the novel concept of a nanochannels photoelectrochemical (PEC) biosensor based on the integration of a unique CuxO-nanopyramid-islands (NPIs) photocathode, an anodic aluminum oxide (AAO) membrane, and alkaline phosphatase (ALP) catalytic chemistry. The CuxO-NPIs photocathode possesses good performance, and further assembly with AAO yields a designed architecture composed of vertically aligned, highly ordered nanoarrays on top of the CuxO-NPIs film. After biocatalytic precipitation (BCP) was stimulated within the channels, the biosensor was used for the successful detection of ALP activity. This study has not only provided a novel paradigm for an unconventional nanochannels PEC biosensor, which can be used for general bioanalytical purposes, but also indicated that the new concept of nanochannel-semiconductor heterostructures is a step toward innovative biomedical applications.

Plasmon-enhanced Raman spectroscopic metrics for in situ quantitative and dynamic assays of cell apoptosis and necrosis.

Apoptosis and necrosis are distinct cell death processes related to many cellular pathways. In situ, quantitatively and dynamically monitoring such processes may provide vitally important information for cell studies. However, such a method still remains elusive, even though current immunochemical methodologies have developed extremely valuable tools. Herein, we demonstrate Raman spectroscopic metrics for validating and quantifying apoptotic and necrotic cells based on their distinct molecular vibrational fingerprints. It not only allows us to quantify apoptotic and necrotic cell populations in situ in adherent cell samples, but also to be capable of continuously monitoring the dynamical processes of apoptosis and necrosis at the same time in one sample. This method provides comparable results with the "gold standard" of flow cytometry, moreover, with several incomparable advantages. Our work offers a powerful new tool for cell apoptosis and necrosis assays and is expected to become a benchmark technology in biological and medical studies.

[Effects of wind speed on drying processes of fuelbeds composed of Mongolian oak broad-leaves.] / é£Žé€Ÿå¯¹è’™å¤æ Žé˜”å¶åºŠå±‚å¤±æ°´è¿‡ç¨‹çš„å½±å“.

Water desorption processes of fuel beds with Mongolian oak broad-leaves were observed under conditions with various wind speeds but nearly constant air temperature and humidity. The effects of wind speed on drying coefficients of fuel beds with various moisture contents were analyzed. Three phases of drying process, namely high initial moisture content (>75%) of phase 1, transition state of phase 2, and equilibrium phase III could be identified. During phase 1, water loss rate under higher wind speed was higher than that under lower wind speed. Water loss rate under higher wind speed was lower than that under lower wind speed during phase 2. During phase 3, water loss rates under different wind speeds were similar. The wind effects decreased with the decrease of fuel moisture. The drying coefficient of the Mongolian oak broad-leaves fuel beds was affected by wind speed and fuel bed compactness, and the interaction between these two factors. The coefficient increased with wind speed roughly in a monotonic cubic polynomial form.

[Effects of polydatin on learning and memory and Cdk5 kinase activity in the hippocampus of rats with chronic alcoholism].

OBJECTIVE: To observe the effects of polydatin on learning and memory and cyclin-dependent kinase 5 (Cdk5) kinase activity in the hippocampus of rats with chronic alcoholism. METHODS: Forty rats were randomly divided into 4 groups: control group, chronic alcoholism group, low and high polydatin group. The rat chronic alcoholism model was established by ethanol 3.0 g/(kg · d) (intragastric administration). The abstinence scoring was used to evaluate the rats withdrawal symptoms; cognitive function was measured by Morris water maze experiment; Cdk5 protein expression in the hippocampus was detected by immunofluorescence; Cdk5 kinase activity in the hippocampus was detected by liquid scintillation counting method. RESULTS: The abstinence score, escape latency, Cdk5 kinase activity in chronic alcoholism group rats were significantly higher than those of control group (P < 0.05). The abstinence score, escape latency in high polydatin group rats were significantly lower than those of chronic alcoholism group (P < 0.05); Cdk5 kinase activity in high and low polydatin group rats was significantly lower than that of chronic alcoholism group( P < 0.05); immunofluorescence showed that the Cdk5 positive cells of chronic alcoholism group were significantly increased compared with control group (P < 0.05), and the Cdk5 positive cells of polydatin groups were significantly decreased compared with chronic alcoholism group ( P < 0.05). CONCLUSION: Polydatin-reduced the chronic alcoholism damage may interrelate with regulation of Cdk5 kinase activity.

Neurosubstrates and mechanisms underlying the extinction of associative motor memory.

Eyeblink conditioning is one of the most commonly used model systems to investigate the neural mechanisms underlying associative motor learning. It is well established that the acquisition and retention of conditioned eyeblink responses (CRs) involve neural plasticity in both the cerebellar cortex and deep cerebellar nuclei (DCN). Nevertheless, how learned CRs are extinguished remains relatively unclear. It has been suggested that extinguished CRs can recur spontaneously, can reappear by exposure to certain stimuli, and can be reacquired in fewer training trials than originally needed, indicating that associative motor memory is not merely erased by extinction training. Instead, the motor memory is preserved to some degree. Herein, we reviewed recent experimental findings demonstrating that the cerebellum subserves the preservation of learned CRs. In addition, several lines of evidence have suggested that forebrain structures (i.e., the medial prefrontal cortex and hippocampus) are involved in the CR extinction. We proposed possible mechanisms related to how preserved motor memory in the cerebellum is inhibited by the forebrain structures via the amygdalar complex.

Synthesis and application of enantioenriched functionalized α-tetrasubstituted α-amino acids from biocatalytic desymmetrization of prochiral α-aminomalonamides.

Catalyzed by Rhodococcus erythropolis AJ270, an amidase-containing microbial whole cell catalyst in neutral phosphate buffer at 30 °C, a number of prochiral α-substituted α-aminomalonamides underwent highly efficient and enantioselective hydrolytic desymmetrization to afford functionalized α-tetrasubstituted α-amino acids in 74-98% chemical yields and 94.0 to >99.5% ee. The presence of a free α-amino (NH(2)) substituent in the substrates was deemed important to ensure high biocatalytic efficiency and enantioselectivity. The synthetic application of biocatalytic desymmetrization was demonstrated by practical chemical transformations of (R)-2-amino-2-carbamoylpent-4-enoic acid to α-substituted serine analogues and a bioactive diamino alcohol derivative.

Excess nicotinamide inhibits methylation-mediated degradation of catecholamines in normotensives and hypertensives.

Nicotinamide and catecholamines are both degraded by S-adenosylmethionine-dependent methylation. Whether excess nicotinamide affects the degradation of catecholamines is unknown. The aim of this study was to investigate the effect of nicotinamide on the methylation status of the body and methylation-mediated catecholamine degradation in both normotensives and hypertensives. The study was conducted in 19 normotensives and 27 hypertensives, using a nicotinamide-loading test (100 mg orally). Plasma nicotinamide, N(1)-methylnicotinamide, homocysteine (Hcy), betaine, norepinephrine, epinephrine, normetanephrine and metanephrine levels before and 5 h after nicotinamide loading were measured. Compared with normotensives, hypertensives had higher baseline (fasting) levels of plasma nicotinamide, Hcy and norepinephrine, but lower levels of plasma normetanephrine, a methylated norepinephrine derivative. Nicotinamide loading induced a significant increase in the levels of plasma N(1)-methylnicotinamide and norepinephrine, and a significant decrease in the levels of O-methylated epinephrine (metanephrine) and betaine, a major methyl donor, in both hypertensives and normotensives. Moreover, nicotinamide-loading significantly increased plasma Hcy levels, but decreased plasma normetanephrine levels in normotensives. The baseline levels of plasma epinephrine in hypertensives were similar to those of normotensives, but the post-nicotinamide-loading levels of plasma epinephrine in hypertensives were higher than those of normotensives. This study demonstrated that excess nicotinamide might deplete the labile methyl pool, increase Hcy generation and inhibit catecholamine degradation. It also revealed that hypertensives had an abnormal methylation pattern, characterized by elevated fasting plasma levels of unmethylated substrates, nicotinamide, Hcy and norepinephrine. Therefore, it seems likely that high nicotinamide intake may be involved in the pathogenesis of Hcy-related cardiovascular disease.

[Expression of LYVE-1 and Prox-1 in non-small cell lung cancer and the relationship with lymph node metastasis].

OBJECTIVE: To investigate the expression of lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), the homeobox gene (Prox-1), in patients with non-small cell lung cancer (NSCLC), the relationship with microlymphatic vessel density, lymph node metastasis and their clinic pathological value. METHODS: Forty specimens of the NSCLC as experimental group and eleven pulmonary benign diseases as control group were studied. The expressions of LYVE-1, Prox-1 and CD31 protein in specimens of NSCLC and normal pulmonary tissue were studied with immunohistochemical (IHC) technique. Microlymphatic vessel density (MLVD) and microvessel density (MVD) were counted. Meanwhile, all specimens were also examined by conventional pathological method. Clinicopathological data of each patient were recorded and analyzed. RESULTS: (1) Among 40 cases of the center of NSCLC cancerous tissues, the MLVDs marked by LYVE-1 and Prox-1 were 4.22 +/- 1.25 and 1. 99 +/- 1.49 respectively, which were significantly lower than those of the pulmonary benign diseases tissues (P = 0.00). (2) The MLVDs marked by LYVE-1 and Prox-1 in NSCLC cancerous invasive edge were 10.89 +/- 2.06 and 6.63 +/- 1.99 respectively, which were significantly higher than those in the center of cancerous tissues and those of the pulmonary benign diseases tissues (P = 0.000). (3) The MLVDs marked by LYVE-1 and Prox-1 in cancerous invasive edge were not correlated with age, gender, site and dimension of lesion, types of histological and degree of differentiation, but correlated significantly with lymph node metastasis (P = 0.000) and PTNM stage (P = 0.000). Meanwhile, along with lymph node metastasis and increasing of PTNM stage, the expressions of LYVE-1 and Prox-1 protein and MLVDs have significantly increased, but the microvessel density marked by CD31 in cancerous invasive edge was not correlated significantly with lymph node metastasis (P = 0.450) and PTNM stage (P = 0.377). (4) Significant correlation between LYVE-1 and Prox-1 (r = 0.529, P = 0.000) expression was observed in NSCLC; moreover, no correlations between LYVE-1 and CD31, Prox-1 and CD31 (r = 0.034, P = 0.837; r = -0.075, P = 0.647) were The functional microlymphatic vessels correlated with lymphatic metastasis are mainly observed. CONCLUSION: located in the cancerous invasive edge rather than the center of cancerous tissues. LYVE-1 and Prox-1 might be acted as molecular phenotypes of lymphangioghesis in NSCLC and as important markers for evaluating lymphatic metastasis and prognosis in patients with NSCLC.

Endogenous reactive oxygen species modulates voltage-gated sodium channels in dorsal root ganglia of rats.

We recently reported that reactive oxygen species (ROS) plays an excitatory role in modulation of the exercise pressor reflex (EPR) in normal rats. In this study, we further tested two independent hypotheses: 1) ROS interacts with EPR-related ionotropic receptors such as the purinergic receptors (P(2)) and transient receptor potential vanilloid 1 receptors (TRPV1) to indirectly modulate the EPR function; 2) ROS directly affects excitability of muscle afferents by modulating the voltage-gated sodium (Na(v)) channels. To test the first hypothesis, we performed animal experiments to investigate the effect of the SOD mimetic 4-hydroxy-2,2,6,6-tetramethyl piperidine 1-oxyl (Tempol) on the pressor response to hindlimb intra-arterial (IA) injection of either α,ß-methylene ATP (a P(2X) agonist) or capsaicin (a TRPV1 agonist) in decerebrate rats. To test the second hypothesis, we used the patch-clamp technique to determine the effect of ROS on Na(v) channels on the soma of muscle afferents. We also performed local microinjection of a sodium channel blocker, tetrodotoxin (TTX), into ipsilateral L4/L5 dorsal root ganglia (DRGs) to investigate whether the blockade of Na(v) channels by TTX affects the EPR function. We found that Tempol did not affect the pressor response to injection of either capsaicin or α,ß-methylene ATP but significantly decreased the Na(v) current in small and medium-sized 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI)-labeled DRG neurons. A membrane-permeant superoxide dismutase, polyethylene glycol (PEG)-SOD, had an effect on the Na(v) current in these neurons similar to that of Tempol. Microinjection of TTX into L4/L5 DRGs dramatically attenuated the pressor response to static contraction induced by electrical stimulation of L4/L5 ventral roots. These data suggest that ROS modulates the EPR by affecting the activity of the Na(v) channels on muscle afferents.

Nicotinamide overload may play a role in the development of type 2 diabetes.

AIM: To investigate whether nicotinamide overload plays a role in type 2 diabetes. METHODS: Nicotinamide metabolic patterns of 14 diabetic and 14 non-diabetic subjects were compared using HPLC. Cumulative effects of nicotinamide and N(1)-methylnicotinamide on glucose metabolism, plasma H(2)O(2) levels and tissue nicotinamide adenine dinucleotide (NAD) contents of adult Sprague-Dawley rats were observed. The role of human sweat glands and rat skin in nicotinamide metabolism was investigated using sauna and burn injury, respectively. RESULTS: Diabetic subjects had significantly higher plasma N(1)-methylnicotinamide levels 5 h after a 100-mg nicotinamide load than the non-diabetic subjects (0.89 +/- 0.13 micromol/L vs 0.6 +/- 0.13 micromol/L, P < 0.001). Cumulative doses of nicotinamide (2 g/kg) significantly increased rat plasma N(1)-methylnicotinamide concentrations associated with severe insulin resistance, which was mimicked by N(1)-methylnicotinamide. Moreover, cumulative exposure to N(1)-methylnicotinamide (2 g/kg) markedly reduced rat muscle and liver NAD contents and erythrocyte NAD/NADH ratio, and increased plasma H(2)O(2) levels. Decrease in NAD/NADH ratio and increase in H(2)O(2) generation were also observed in human erythrocytes after exposure to N(1)-methylnicotinamide in vitro. Sweating eliminated excessive nicotinamide (5.3-fold increase in sweat nicotinamide concentration 1 h after a 100-mg nicotinamide load). Skin damage or aldehyde oxidase inhibition with tamoxifen or olanzapine, both being notorious for impairing glucose tolerance, delayed N(1)-methylnicotinamide clearance. CONCLUSION: These findings suggest that nicotinamide overload, which induced an increase in plasma N(1)-methylnicotinamide, associated with oxidative stress and insulin resistance, plays a role in type 2 diabetes.

[Expression of neuronal nitric oxide synthase in corpus cavernosum with diabetic erectile dysfunction: experiment with rabbits].

OBJECTIVE: To investigate the expression characteristics of neuronal nitric oxide synthase (nNOS) in the corpus cavernosum of diabetic erectile dysfunction (ED). METHODS: Fifty rabbits underwent single intravenous injection of alloxan (100 mg/kg). Blood glucose, system arterial pressure (SAP), and intracavemosal pressure (ICP) were measured 15, 30, and 45 days later. The rabbits with the blood glucose >10 mmol/L were used as diabetic models and those with the ICP lower than the normal control group by more than 25% were used as diabetic ED models. RT-PCR, immunohistochemistry, and Western-blotting were used to detect the expression of nNOS in the corpus cavemosum. RESULTS: Seven days after the alloxan injection the blood glucose levels of 35 rabbits (70%) were > 10 mmol/L. 40% rabbits (14/35) were were used as diabetic ED models. Compared with control group, the ICP 15, 30, and 45 days after alloxan injection were considerately decreased in diabetic ED rabbits (all P < 0.01), and there was no significant difference in SAP between the two groups (all P > 0.05). The relative values of nNOS mRNA expression 15, 30, and 45 days after alloxan injection of diabetic ED rabbits were 0.670 +/- 0.030, 0.451 +/- 0.012, and 0.206 +/- 0.023 respectively,all significantly lower than those of the control group (0.817 +/- 0.010, 0.814 +/- 0.020, and 0.802 +/- 0.007 respectively, all P < 0.05). Western blotting showed that the relative values of nNOS 15, 30, and 45 days after alloxan injection of the ED rabbits were 0.713 +/- 0.014, 0.424 +/- 0.007, and 0.337 +/- 0.009 respectively, all significantly lower than those of the control group (0.797 +/- 0.015, 0.706 +/- 0.020, and 0. 750 +/- 0.022 respectively, all P < 0.05). CONCLUSION: Decreased expression of nNOS in corpus cavernosum is one of possible pathway of diabetic ED.