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Objective@#To investigate whether γ-aminobutyric acid (GABA) receptor signaling pathway is involved in the regulation of thalamic undefined (ZI)-nucleus accumbens (NAc) neural pathways on gastric distraction (GD)-sensitive neuronal firing activity and the impact on food intake, the number of times and the frequency in rats.@*Methods@#Six rats were randomly selected and the neural pathway between ZI and NAc in rat thalamus was observed by fluorescent gold (FG) retrograde tracing method.Eighty-two rats were randomly selected, and the gastric balloon was placed in gastric cavity, the microelectrode was placed in the NAc, and the stimulating electrode was placed in the ZI. The single-cell discharge recording method was used to observe the effect of electrical stimulation ZI on the excitability of GD-sensitive neurons in rat NAc.Eighteen rats were randomly selected and were divided into three groups according to the random number table. They were NS group, GABA group, GABA + GABA receptor antagonist bicuculline (BIC) group with 6 in each group, and the rat NAc was used to embed the cannula. The method of GABA and BIC was injected to observe the changes of cumulative food intake in rats for 4 h. Eighteen rats were randomly selected and randomly divided into three groups: sham stimulation (SS) group, 50 μA electrical stimulation group, 50 μA electrical stimulation + BIC group with 6 in each group. The 4 h cumulative food intake of rats was observed by electro-stimulation of rat ZI and rat NAc injection of BIC.@*Results@#Fluorescent gold retrograde tracking combined with fluorescent immunohistochemical staining showed that there were visible GABA and fluorescent gold double labeled neurons in ZI. Electrical stimulation of ZI, the frequency of GABA-sensitive GD neurons in rat NAc increased significantly (GD-E increase: (78.8±8.4)%, GD-I increase: (89.3±9.2)%, P<0.01), but the inhibitory effect was antagonized by BIC (GD-E increase: (113.8±13.6)%, GD-I increase: (121.8±14.2)%, P<0.01). Microinjection of GABA in rat NAc significantly increased the cumulative food intake for 4 h ((155.72±18.84) kcal, t=3.41, P<0.05), which was antagonized by partial BIC (123.43±15.11) kcal, t=3.28, P<0.05). Electrical stimulation of ZI significantly increased the food intake in rats ((39.07±11.27) kcal, t=2.96, P<0.05), and this effect can be partially antagonized by BIC ((34.17±10.85)kcal, t=2.33, P<0.05).@*Conclusion@#The ZI-NAc neural pathway regulates the discharge activity of rat gastric distension (GD)-sensitive neurons and the feeding status of rats, and the GABA receptor signaling pathway may be involved in mediating the process.
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@# Objective: To investigate the role and mechanism of Krüppel-like factor 4 (KLF4) in regulating epithelial-mesenchymal transition (EMT) and migration of bladder cancer cells. Methods: Bladder cancer 5637 and T24 cell lines that stably over-expressing KLF4 (LV-KLF4, experiment group) were constructed, and the negative control group (LV-NC) was also established; the mRNA and protein expressions of KLF4 were verified by qPCR and WB, respectively. Transwell chamber assay was used to detect the migration ability of cells in LV-KLF4 and LV-NC groups. WB was performed to detect the expression levels of EMT-related markers (E-cadherin, N-cadherin, Vimentin) and Wnt signaling pathway-related proteins. Immunofluorescence technique was used to detect the distribution of β-catenin in cells after over-expression of KLF4. Results: The 5637 and T24 cell lines over-expressing KLF4 gene were successfully constructed. Compared with the LV-NC group, the mRNA and protein expressions of KLF4 increased in LV-KLF4 groups (all P<0.01); the expression of E-cadherin increased (P<0.01), while the expressions of N-cadherin, vimentin, and the expression levels of total β -catenin, nuclear β -catenin, MMP 9 and c-Myc decreased (all P<0.01); moreover, the migration ability of cells decreased significantly (P<0.01); the fluorescence expression of β-catenin in cells also decreased significantly in LV-KLF4 group as compared to LV-NC group. Conclusion: Over-expression of KLF4 gene in bladder cancer cells may inhibit EMT process by regulating Wnt/β-catenin signaling pathway, and further inhibit the migration of bladder cancer 5637 and T24 cells.
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Objective To investigate whether γ-aminobutyric acid (GABA) receptor signaling pathway is involved in the regulation of thalamic undefined (ZI)-nucleus accumbens (NAc) neural pathways on gastric distraction (GD)-sensitive neuronal firing activity and the impact on food intake,the number of times and the frequency in rats.Methods Six rats were randomly selected and the neural pathway between Zl and NAc in rat thalamus was observed by fluorescent gold (FG) retrograde tracing method.Eighty-two rats were randomly selected,and the gastric balloon was placed in gastric cavity,the microelectrode was placed in the NAc,and the stimulating electrode was placed in the ZI.The single-cell discharge recording method was used to observe the effect of electrical stimulation ZI on the excitability of GD-sensitive neurons in rat NAc.Eighteen rats were randomly selected and were divided into three groups according to the random number table.They were NS group,GABA group,GABA + GABA receptor antagonist bicuculline (BIC) group with 6 in each group,and the rat NAc was used to embed the cannula.The method of GABA and BIC was injected to observe the changes of cumulative food intake in rats for 4 h.Eighteen rats were randomly selected and randomly divided into three groups:sham stimulation (SS) group,50 μA electrical stimulation group,50 μA electrical stimulation + BIC group with 6 in each group.The 4 h cumulative food intake of rats was observed by electro-stimulation of rat ZI and rat NAc injection of BIC.Results Fluorescent gold retrograde tracking combined with fluorescent immunohistochemical staining showed that there were visible GABA and fluorescent gold double labeled neurons in ZI.Electrical stimulation of ZI,the frequency of GABA-sensitive GD neurons in rat NAc increased significantly (GD-E increase:(78.8±8.4) ％,GD-I increase:(89.3±9.2) ％,P＜0.01),but the inhibitory effect was antagonized by BIC (GD-E increase:(113.8 ± 13.6)％,GD-I increase:(121.8± 14.2)％,P＜0.01).Microinjection of GABA in rat NAc significantly increased the cumulative food intake for 4 h ((155.72± 18.84) kcal,t=3.41,P＜0.05),which was antagonized by partial BIC (123.43± 15.11) kcal,t =3.28,P＜ 0.05).Electrical stimulation of ZI significantly increased the food intake in rats ((39.07± 11.27) kcal,t =2.96,P＜0.05),and this effect can be partially antagonized by BIC ((34.17 ± 10.85) kcal,t =2.33,P＜ 0.05).Conclusion The ZI-NAc neural pathway regulates the discharge activity of rat gastric distension (GD)-sensitive neurons and the feeding status of rats,and the GABA receptor signaling pathway may be involved in mediating the process.
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Objective To investigate the effects of orexin-A on firing activity of gastric distensionsensitive (GD) neurons in the basomedial amygdala (BMA) and food intake in diet-indaced obese rats.Methods Healthy male Wistar rats were selected,and the diet-induced obesity (DIO) rat model and dietinduced resistant (DR) rat model were established by high-fat diet.The effects of orexin-A and an opioid receptor antagonist naloxone on BMA GD neurons were observed by recording the extracellular potentials of single neurons.The effects of orexin-A and naloxone on the food intake of different rats were observed by using BMA catheterization.The mRNA expression and protein expression of orexin-1 receptor (OX-1R) and μ opioid receptor were detected by real-time PCR and Elisa,respectively.Results After microinjection of orexinA into the BMA,the firing frequency of GD-sensitive neurons in the normal rats was significantly increased (GD-E:(78.3±6.9)％,GD-Ⅰ:(55.5±4.7) ％,P＜0.01),and this effect was completely blocked by OX-1R receptor antagonist SB334867,and naloxone partially blocked the discharge-promoting effect of orexin-A;Compared with the normal rats,the firing frequency of GD-sensitive neurons in the DIO (GD-E:(91.6±7.1) ％,GD-Ⅰ:(67.9±8.1) ％) and DR(GD-E:(87.9±6.8) ％,GD-Ⅰ:(69.2±5.8) ％) rats was significantly increased after BMA injection of orexin-A (P＜0.05).After administration of orexin-A into the BMA,food intake of the normal rats,DIO rats and DR rats ((2.38±0.34) g,(3.75 ±0.32) g,(4.01 ±0.38) g,respectively) was significantly increased (P＜0.01),and the food intake of DR and DIO rats were significantly higher than that of normal rats (P＜0.05).After BMA was injected with naloxone,the food intake of rats was inhibited,and the food intake of the DIO rats was significantly lower than that of the DR rats (P＜0.05),food intake of the DR rats was significantly lower than that of the normal rats (P＜0.05).The results of real-time PCR showed that the mRNA levels of OX-1R in DIO and DR rats were(5.85±0.45)and (6.03±0.42)were higher than that of normal rats,and the difference was significant (P＜0.05);and mRNA levels of μ-opioid receptors in DIO and DR rats((4.51±0.42) and (8.31±0.41) times) were higher than those in normal rats (P＜0.05).The results of Elisa showed that the protein levels of OX-1R in DIO ((2.98±0.28) ng/μl)and DR rats ((3.05±0.31) ng/μl) were higher than those in normal rats ((1.53±0.31) ng/μl,P＜0.05).The content of μ-opioid receptor protein in DR rats ((4.21±0.35) ng/μl) was higher than that of DIO rats ((2.77±0.27) ng/μl),and higher than that of normal rats((1.48±0.32) ng/μ),the difference was significant (P＜0.05).Conclusion BMA orexin-A promotes the spontaneous discharge of GD-sensitive neurons and food intake in normal rats,DIO rats and DR rats,μ-opioid receptors may be involved in the regulation of this process.
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Objective:This study aimed to explore the Ventromedial Hypothalamic Orexin-1 and Orexin-1 Receptors in Regulation of Gastric Acid Secretion in Conscious Rats.Methods:Rats were anaesthetized and fitted with a stainless steel carmula placed just above the VMH or paracele,after random allocation orexin-A,[Pro34]-peptide YY and [CPP1-7,NPY19-23,Ala31,Aib32,Gln34] -pancreatic polypeptide were injected in the VMH;SB-334867 was intraperitoneal injection;atropine was subcutaneous injection;GR-231118 and CGP-71683 were injected in the paracele.Using pyloric ligation model,tests the effect of different drugs on rat gastric acid secretion and gastric juice volume.Results:OXA induced dose-dependent increase of gastric acid secretion;SB-334867 induced dose-dependent inhibition of gastric acid secretion.The stimulatory effect of OXA on acid secretion was inhibited by SB-334867;atropine induced dose-dependent increase of gastric acid secretion and block the effect of orexin-A on gastric acid secretion;the gastric acid secretion was inhibited by GR-231118 or CGP-71683,and GR-231118 or CGP-71683 were blocked the effect of orexin-A on gastric acid secretion;Intraventromedial hypothalamic injections of [CPP1-7,NPY19-23,Ala31,Aib32,Gln34]-pancreatic polypeptide increased gastric acid secretion.Conclusion:It is suggested that endogenous orexin-A acts on the ventromedial hypothalamus to stimulates acid secretion.This stimulatory effect is probably mediated through orexin receptor,Y1 and Y5 receptor,and the vagus nerve system.
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Objective:To investigate the effects of Orexin peptides on feeding and energy metabolism in mice.Methods:The mice were divided into two groups:feeding group and metabolic group.The feeding group were injected with different doses (1,3 and 10 nmol) of orexin-A and orexin-B to observe their effects on feeding and the activity of tyrosine hydroxylase in liver.We used the metabolic cages and observed the changes of respiration rate and respiration rate of mice were under light condition,dark condition and fasting condition.Results:Compared with the control group,1 nmol and 10 nmol orexin-A significantly stimulated mice to feed (P ＜0.05) within 4 hours after injection,and the effect of 3 nmol orexin-A on feeding was not obvious,but increase the activity of tyrosine hydroxylase.Any dose of orexin-B did not show a stimulating effect on mice feeding.(P ＞0.05).In the light cycle,orexin-A could significantly reduce the respiration rate (RQ),the metabolic rate was significantly increased (P ＜0.05);In the dark cycle,orexin-A had no effect on RQ,but the metabolic rate was significantly rised (P ＜0.05);But the injection of orexin-A in fasting mice induced a brief increase in RQ and a significant increase in metabolic rate (P ＜0.05).Conclusion:Orexins may play an important role in regulating feeding and energy metabolism in mice.
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Objective:The current study investigated the effects of nesfatin-1 in the hypothalamic paraventricular nucleus (PVN) on gastric motility and the regulation of the lateral hypothalamic area (LHA).Methods:The projection of nerve ?ber and expression of nesfatin-1 were observed by retrograde tracing and fluo-immunohistochemistry staining;The nuclei microinjection and nuclei electrical stimulation,extracellular discharges of single unit neuron were used to observe the effects of nesfatin-1 on the GD neurons;Gastric motility recording in vivo were used to monitor the effects of nesfatin-1 on the amplitude of constriction and frequency of gastric motility in conscious rats.Results:Nesfatin-1 inhibited the majority of the GD-E neurons(1.97± 0.12 Hz vs.1.15± 0.07 Hz) and excited GD-I neurons (1.74± 0.10 Hz vs.3.04± 0.18 Hz) in the PVN,which were weakened by oxytocin receptor antagonist H4928 (GD-E:1.38± 0.08 Hz,P＜0.05 vs.nesfatin-1;GD-I:2.49± 0.15 Hz,P＜0.05 vs.nesfatin-1).Gastric motility experiments showed that administration ofnesfatin-1 in the PVN decreased gastric motility.Retrograde tracing and immunofluorescent staining showed that nucleobindin-2/nesfatin-1 and fluorogold double-labeled neurons were observed in the LHA.Electrical LHA stimulation excited the firing rate of GD-responsive neurons (GD-E:2.06± 0.12 Hz vs.4.23± 0.21 Hz,GD-I:1.61± 0.09 Hz vs.4.83± 0.25 Hz) in the PVN.Pre-administration of an antinucleobindin-2/nesfatin-1 antibody in the PVN strengthened gastric motility,decreased GD-E neurons (1.74± 0.10 Hz vs.3.04± 0.18 Hz) and excited the discharging of the GD-I neurons(4.15± 0.18 Hz vs.4.83± 0.25) induced by electrical stimulation of the LHA.Conclusion:Nesfatin-1 in the PVN could serve as an inhibitory factor to inhibit gastric motility,which might be regulated by the LHA.
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Objective:This study aimed to explore the effects ofnesfatin-1 on gastric distension (GD)-sensitive neurons in the basomedial amygdala (BMA) and the potential mechanism for nesfatin-1 to regulate gastric motility through the arcuate nucleus (Arc).Methods:The projection of nerve fiber and expression of nesfatin-1 were observed by retrograde tracing and fluo-immunohistochemistry staining;The nuclei microinjection and nuclei electrical stimulation,extracellular discharges of single unit neuron were used to observe the effects ofnesfatin-1 on the GD neurons;Gastric motility recording in vivo were used to monitor the effects ofnesfatin-1 on the amplitude of constriction and frequency of gastric motility in conscious rats.Results:NUCB2/Nesfatin-1/fluorogold-double labeled neurons were from ARC to BMA;Nesfatin-1 could excited the firing rate of most of the GD-E neurons (4.25± 1.02 Hz vs.5.32± 1.17 Hz,P＜0.01) and decreased the firing rate of most of the GD-I neurons (3.73± 0.92 Hz vs.2.64± 0.86 Hz,P＜0.01),inhibited the gastric motility,amplitude and frequency,SHU9119 could weaken the responses induced by nesfaton-1;Electrical stimulation of the Arc,the firing rate of nesfatin-1-induced GD-response neurons (GD-E:5.14± 1.32 Hz vs.6.75± 1.84 Hz,P＜0.05;GD-I:2.84± 0.86 Hz vs.4.05± 1.12 Hz,P ＜0.05) and the gastric amplitude and frequency were increase.Conclusion:It was suggested that nesfatin-1 in the BMA plays an important role in decreasing gastric motility and the Arc may be involved in this regulation process.
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OBJECTIVE TNF- related apoptosis- inducing ligand(TRAIL)is a promising cancer therapeutic agent due to its minimal toxicity to normal tissues and remarkable apoptotic activity in tumors. However, most breast cancer cells are resistant to TRAIL- induced apoptosis. Our objectives are to investigate the underlying molecular mechanisms and to develop strategies to overcome such resistance. METHODS To identify modulators of TRAIL-induced apoptosis, we carried out a genome wide siRNA screen. To validate the screening result, we either silenced or overexpressed the identified genes in various breast cancer cells and changes in growth and TRAIL-induced cell apoptosis were determined in vitro and in an orthotopic xenograft mouse model. Finally, we investigated whether small molecules targeting the identified genes improve the effectiveness of TRAIL-therapy. RESULTS We unexpectedly identified androgen receptor (AR) to be responsible for TRAIL resistance. While AR is classically viewed as the key factor in prostate cancer progression, we found that AR expression levels were markedly elevated in human invasive breast cancer specimens including triple- negative breast cancers (TNBC) that are highly aggressive with poor prognosis. Importantly, breast cancer cell lines express different levels of AR that correlated with their TRAIL resistance. AR overexpression in MDA- MB- 231 and MDA- MB- 436 cells suppressed the TRAIL sensitivity whereas knockdown of AR rendered MCF-7 and MDA-MB-453 cells sensitive to TRAIL-induced apoptosis. AR overexpression also induced TRAIL resistance in breast tumors in vivo. Further, we observed an upregulation of the TRAIL receptor, death receptor 5 (DR5) in breast cancer cells, following the removal or inhibition of AR by its antagonists Casodex and MDV3100. Treatment with AR antagonists also enhanced TRAIL- induced breast cancer cell apoptosis. CONCLUSION AR signaling suppresses TRAIL-induced breast cancer cell apoptosis, in part, by suppressing DR5 expression, and a combination of AR antagonists together with TRAIL may be a novel and effective therapy for TNBC.
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Objective To observe the protective effect of ghrelin on hippocampal injury induced by global cerebral ischemia/reperfusion (I/R) and explore its effect mechanisms.Methods The male Sprague-Dawley (SD) rats were randomly divided into four groups,namely sham group,I/R group,normal saline (NS)+I/R group and Ghrelin+I/R group,with 42 rats in each group.The model of I/R was reproduced by clipping bilateral carotid artery of rats 15 minutes and then releasing them for 60 minutes.There were no challenges for rats in sham group,just exposed their carotid artery.Ghrelin+I/R group and NS+I/R group were challenged by injecting 1 μ.L ghrelin or NS into lateral ventricle before I/R.Some of brain tissue in the rats was harvested after experiment to determine the levels of malonaldehyele (MDA),myeloperoxidase (MPO) and glutathione (GSH) in hippocampus by using chemical colorimetry and observe infarct sizes and histopathology.Single extracellular neuron discharge in other rats was recorded to observe the activity of glutamic sensitive neurons (Glu-N) and γ-aminobutyric acid (GABA) sensitive neurons (GABA-N) in hippocampus CA1 region of rats suffered I/R.Results Compared with sham group,the levels of MDA and MPO in hippocampus of rats in the I/R group were raised markedly,the level of GSH was decreased significantly,the infarct sizes was increased significantly and pycnosis neurons were increased markedly.All sorts of indexes between NS+I/R group and I/R group showed no significantly statistical significance.Compared with NS+I/R group,the levels of MDA and MPO in hippocampus of rats in the Ghrelin+I/R group were decreased significantly [MDA (nmol/g):16.4 ± 4.2 vs.24.5 ± 6.7,MPO (nmol/g):6.4 ± 1.8 vs.10.2 ± 2.9,both P ＜ 0.05],the activity of GSH was risen remarkably (μmol/g:2.65 ± 0.72 vs.1.66 ± 0.50,P ＜ 0.05),the infarct sizes of hippocampus were reduced markedly [(43.9 ± 9.5)％ vs.(77.0 ± 12.7)％,P ＜ 0.01],the number of pycnosis neuron was reduced markedly (cells:36.2±4.5 vs.47.1 ±6.1,P ＜ 0.01).The results of electrophysiology showed that the discharge frequency of Glu-N and GABA-N in hippocampus CA1 region of rats in I/R group increased markedly as compared with sham group,and no significant difference in the discharge frequency of Glu-N and GABA-N between NS+I/R group and I/R group.Compared with NS+I/R group,injected ghrelin could make the discharge frequency of Glu-N in hippocampus CA1 region of rats decreased markedly (Hz:3.81 ±0.67 vs.4.98±0.33 at ischemia,3.01 ±0.37 vs.3.77 ± 0.41 at reperfusion,both P ＜ 0.05),and the discharge frequency of GABA-N increased markedly (Hz:5.62 ± 0.54 vs.3.62±0.39 at ischemia,4.81±0.48 vs.3.71±0.21 at reperfusion,both P ＜ 0.05).Conclusion Ghrelin might protect hippocampal neuron after I/R iniury,and neuron excitability decrease might be related.
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Objective To study the effect of Ghrelin on food choice of rats.Methods Twenty-four Wistar rats were randomly divided into control group,0.1 nM Ghrenlin group and 1.0 nM Ghrenlin group,and 8 rats in each group.0.2 μl arificial cerebrospinal fluid were injected into the lateral ventricle in control group,and the same volume 0.1nM and 1.0 nM Ghrelin were done in 0.1 nM Ghrenlin group and 1.0 nM Ghrenlin group respectively.The changes of food intake,feeding interval,and torlerance of different flavors of food intake were observed.The SCH23390,an antagonist of dopamine D1 receptor,was used to explore the mechanisms of Ghrelin.Results The rats' food consumption increased significantly and the intake intervals reduced dose dependently after injecting Ghrelin into the lateral ventricles (P＜0.05 ～ 0.01).Compared with normal liquid food,the rats' intake of food added with quinine was reduced((16.73±5.21)ml vs (23.47±9.46)ml,P＜0.01),and injecting Ghrelin into the lateral ventricles could not reverse this effect ((13.74±3.29) ml vs (16.73±5.21)ml,P＞0.05).Mter injecting Ghrelin,the rats' intake of liquid food added with sugar increased in a dose dependent manner(P＜0.05 ～ 0.01),and higher than the food intake of fasted rats ((59.24 ± 17.32) ml vs (38.13 ± 10.98) ml,P＜ 0.05).The food intake reduced obviously after injecting SCH23390 ((22.69±6.54) ml vs (28.21±7.35)ml,P＜0.05).But the rats' food intake was significantly lower after injecting SCH23390 ± Ghrehn than the rats only injected of Ghrehn ((3Z44±10.62)ml vs (65.81±13.47)ml,P＜0.05).Conclusion Ghrelin can affect the food choice of rats,and dopamine may be involved in the regulation of this process.
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Objective To explore the effects and mechanisms of nesfatin-1 injection in the central nucleus of the amygdale(CeA) on exploratory behavior in rats.Methods Fifty rats were divided into 5 groups randomly : nesfatin-1 low dose group, nesfatin-1 high dose group, SHU9119 group, SHU9119+nesfatin-1 low dose group and control group.Drugs were administrated via CeA to examine behavioral changes of rats by elevated plus maze model test and nesfatin-1 mRNA expression in CeA of anxiety rats (10 rats).Results Compared with the control group,anxiety behaviors of rats in nesfatin-1 low dose group and nesfatin-1 high dose group were improved remarkably,showing that the open arm entries were significantly reduced (OE, t=4.16-12.87, P＜0.01), open arm entry proportion was decreased remarkably (OE％, t=2.39-4.39, P＜0.01-0.05), and time proportion in open arm was decreased significantly (OT％, t=5.43-20.55, P＜0.01),which presented dose dependent.Compared with the control group, anxiety degree of rats in nesfatin-1 low dose group and nesfatin-1 high dose group significantly enhanced, showing that the rats head dipping reduced obviously (HD, t=6.97-16.73, P＜0.01) ,while rearing was increased significantly (RE, t=6.60-13.25, P＜0.01) ,which was presented dose dependent.Melanocortin receptor antagonist SHU9119 can partly eliminate nesfatin-1 induce-anxiety (t =2.11-3.08, P＜ 0.01-0.05).Real-time PCR results showed that nesfatin-1 mRNA expression of anxiety model rats was significantly increased (t =3.40-5.58, P＜0.05).Conclusion Amygdala nesfatin-1 can increase anxiety and fear of rats,which may be associated with melanocortin systems.
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<p><b>OBJECTIVE</b>To study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).</p><p><b>METHODS</b>We established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.</p><p><b>RESULTS</b>The prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.</p><p><b>CONCLUSION</b>The ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.</p>
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Animals , Humans , Male , Rats , Apomorphine , Carcinogens , Diethylstilbestrol , Erectile Dysfunction , Myocytes, Smooth Muscle , Nitric Oxide Synthase Type I , Metabolism , Organ Size , Penile Erection , Penis , Pituitary Neoplasms , Prolactin , Blood , Prolactinoma , Rats, Wistar , Testosterone , BloodABSTRACT
The present study was aimed to investigate the effects of orexin-A and orexin-1 receptor (OX1R) antagonist injected into the fourth ventricle of rats on food-intake and spontaneous physical activity (SPA). Obese rat model was induced by high fat diet. Different doses of orexin-A or SB334867, an OX1R antagonist, were injected into the fourth ventricle of obese and normal rats respectively. SPA and food intake were monitored for 4 h after injection in both light and dark environment. In the light measurement cycle, different doses of orexin-A significantly stimulated feeding and SPA in all injected rats, and the animals' responses showed a dose-dependent manner (P < 0.05-0.01), and compared with those of normal rats, the orexin-A induced food intake and SPA were more pronounced in obese rats. In the dark measurement cycle, different doses of orexin-A had no obvious effect on food intake and SPA in both normal and obese rats (P > 0.05). In the light cycle, different doses of SB334867 significantly decreased food intake and SPA in all rats during 0-2 h and 2-4 h after injection (P < 0.05), but the food intake and SPA in obese rats were significantly greater than those of normal rats. In the dark cycle, different doses of SB334867 showed no obvious effect on food intake and SPA of normal and obese rats (P > 0.05). These results suggest that fourth cerebral ventricle nuclei may be one target for orexin-A and light condition may play an important role in orexin-A and OX1R physiological functional processes.
Subject(s)
Animals , Rats , Benzoxazoles , Pharmacology , Diet, High-Fat , Eating , Fourth Ventricle , Motor Activity , Obesity , Orexin Receptor Antagonists , Pharmacology , Orexin Receptors , Orexins , Pharmacology , Urea , PharmacologyABSTRACT
Objectives To analyze the expression of fibroblast growth factor-19(FGF-19) in hepatocellular carcinoma ( HCC) and adjacent tissues , and to investigate its clinical significance .Methods A total of 209 HCC patients who had undergone radical resection operations at Hospital 401 between January 2003 and December 2009 were chosen as samples . Immunohistochemistry method was employed to examine the expression level of FGF-19 in HCC and adjacent tissues .The relationship between FGF-19 protein expressions and clinicopathological features was analyzed by the chi -square test or Fisher exact probability .A survival curve was drawn using the Kaplan-Meier method and the Cox model was used to analyze factors that influenced survival .Results The rate of high expression of FGF-19 was 66.1% (138/209) in HCC, which was significantly higher than 46.9%(98/209) in adjacent tissues (P<0.05).The high expression of FGF-19 was related to the tumor capsule and tumor boundary (P<0.05).The overall survival in high expression of FGF-19 group was signifi-cantly lower than that in low expression group (P<0.05).Conclusion FGF-19 plays an important role in the carcinogen-esis and development of HCC , and a high expression of FGF-19 might be closely related to survival time of postoperative patients.FGF-19 might be a potential prognosis prediction factor for HCC .
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This study is to investigate the anticancer effects and mechanisms of Tegillarca granosa Linnaeus-1 [TG-1] on renal carcinoma OS-RC-2 cells in vitro. The proliferation of OS-RC-2 cells was evaluated under various concentrations of TG-1 using MTT assay. The apoptosis of OS-RC-2 cells was analyzed using acridine orange/ethidium bromide staining. And the cell cycle distribution of OS-RC-2 cells was detected by flow cytometry. In addition, the expression level of Ki67 mRNA was examined by RT-PCR and level of casepase-3 was examined by Western blot analysis. TG-1 incubation significantly inhibited the proliferation of renal carcinoma OS-RC-2 cells and arrested cells at G[0]/G[1] phase [P <0.05]. And TG-1 also significantly inhibited the expression of Ki67 mRNA [P<0.05]. Additionally, TG- 1 significantly promoted apoptosis and the expression of caspase-3 in cells [P<0.05]. Moreover, the optimal effects of TG-1 was achieved at the concentration of 100 mg/L The results indicate that TG-1 has antitumor effects on renal carcinoma OS-RC-2 cells and that the underlying mechanisms may be acted through inhibiting proliferation and Ki67 mRNA expression, and promoting apoptosis and caspase-3 expression.
Subject(s)
Peptides , Carcinoma, Renal Cell , Ki-67 Antigen , Caspase 3 , Apoptosis , Polymerase Chain Reaction , Blotting, WesternABSTRACT
Objective To research the functional role of thyroidal motilin and the effects of electric excitation of the paraventricular nuclei(PVN) on gastric motility and the levels of motilin in thyroid and plasma.Methods The expression of motilin in rat and human thyroid was detected by immunofluorescence staining.A phase Ⅲ-like contraction was recorded before and after thyroidectomy and after PVN excitation.The changes in concentrations of plasma FT3,FT4 and motilin were determined via radioimmunoassay (RIA).c-Fos expression of PVN after thyroidectomy and motilin expression in thyroid after PVN excitation were observed by immunohistochemical staining.Results There were motilin immunoreactive cells in rat and human thyroid.The phase Ⅲ-like contraction and concentration of motilin in plasma decreased significantly when measured on the second and fourth days after thyroidectomy(2d,P＜0.01 ;4d,P＜0.05).The expression of c-Fos in PVN after thyroidectomy was significantly increased(P＜0.05).An electric excitation of PVN could increase the concentration of motilin in plasma and thyroid and increase corresponding gastric motility in rats (P ＜0.05).The increased phase Ⅲ-like contraction by PVN excitation could be partially inhibited by administration of motilin receptor antagonist,GM-109 (P＜0.05).Excitation of PVN in thyroidectomized rats resulted in lower plasma motilin and less intense phase Ⅲ-like contraction of stomach,as compared with the sham operated control group(P＜0.05).Conclusion Motilin from the thyroid may be secreted into the peripheral plasma to affect gastric motility and PVN may modulate gastric motility and motilin expression in the thyroid.
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Objective To investigate the tracking neurofibra pathway from the hippocamlpal neurons to septal nucleus,and to explore the effects and mechanisms of ghrelin on the learning and memory in septal nucleus lesion rats.Methods Retrogradely tracing method was used to observe Fluorogold (FG) reaching sites in hippocampus.The septal nucleus was destructed by direct current using stereotactic technique.Step-down test and morris water maze were used to test the effects of learning and memory ability by means of microinjecting ghrelin into hippocampal CA1 area in rats.Results After injection of FG into septal nucleus,retrogradely labeled neurons and neurofibra were found in the hippocampal neurons with FG.Ghrelin injection of hippocampal CAI area could promoter learning and memory ability in rats.It showed that the escaped latent period was significantly lengthened ( (3.2 ± 0.9) s vs (6.9 ± 1.1 ) s,P ＜ 0.05) and the wrong numbers in 5 min were obviously decreased in escape response test; and the latency of looking for the plat was significantly shorter in morris water maze test ( 1.8 ±0.4vs 0.8 ± 0.1,P ＜ 0.05 ).However the effects above-mentioned on learning and memory was significantly weak after septal nucleus lesioning.It showed that the escaped latent period was markedly shortened ( ( 19.5 ±3.2)s vs ( 10.5 ± 2.1 ) s,P ＜ 0.05 ) and the wrong numbers in 5 min were obviously increased ( ( 3.9 ± 0.8 ) s vs ( 1.8 ±0.5 ) s,P＜0.05 ) in escape response test.The latency of looking for the plat was significantly lengthened in morris water maze (P＜0.05).Conclusion Ghrelin could elevate the learning and memory ability in hippocampus,and the effects may be related to the septal-hippecampus pathway.
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<p><b>OBJECTIVE</b>To investigate the expression of motilin and its precursor mRNA in normal human thyroid. To compare the expression differences of motilin and it precursor mRNA between normal thyroid and intestines. To study the expression of motilin and its precursor mRNA in human thyroid tumors and their clinical implications.</p><p><b>METHODS</b>RT-PCR, Southern blot and molecular cloning were used to detect motilin transcript expression in human thyroid and mucous membrane of small intestine. Real-time PCR and immunohistochemical techniques were used to quantify motilin precursor mRNA and motilin peptide in thyroid tissue samples including adenoma, medullary carcinoma, follicular carcinoma, papillary carcinoma and nodular goiter.</p><p><b>RESULTS</b>(1) The expression of motilin and its precursor mRNA in normal human thyroid was primarily in the thyroid C cells. (2) RT-PCR and Southern blot showed that motilin mRNA expressed in human thyroid was identical to that expressed in duodenum with identical sequence deposited in NCBI Genbank of America. (3) Immunohistochemistry, Western blot research and real-time PCR studies showed that motilin and its precursor mRNA were expressed in normal and tumor tissues of human thyroid. Thyroid tumors (acidophilic adenoma, medullary carcinoma, follicular carcinoma, papillary carcinoma and nodular goiter) showed intense and diffuse immunostaining for motilin peptide. Moreover, the expression of motilin and its precursor mRNA in thyroid medullar carcinoma and acidophilic adenoma were significantly higher than those of normal thyroid tissue (P < 0.05). The expression in thyroid follicular and papillary carcinomas were significantly lower than those of normal thyroid tissue (P < 0.05). There was no difference of the expression between nodular goiter and normal thyroid tissue (P > 0.05).</p><p><b>CONCLUSIONS</b>Motilin peptide and its precursor mRNA are expressed in C cells of human thyroid. The sequence of motilin is identical to that expressed in duodenum from NCBI Genbank of America. The expressions of both motilin and its precursor mRNA in thyroid medullary carcinoma and acidophilic adenoma are significantly increased. In contrast, their expressions in thyroid follicular and papillary carcinomas are significantly decreased. Motilin may regulate physiological functions of the thyroid through parafollicular cells. Motilin may be involved in the pathogenesis of medullary carcinoma and acidophilic adenoma of the thyroid.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Adenocarcinoma, Follicular , Genetics , Biomarkers, Tumor , Metabolism , Carcinoma, Medullary , Genetics , Carcinoma, Papillary , Genetics , Metabolism , Intestines , Metabolism , Motilin , Genetics , Metabolism , Nervous System Neoplasms , Metabolism , RNA Precursors , Metabolism , RNA, Messenger , Metabolism , Thyroid Gland , Metabolism , Thyroid Neoplasms , Genetics , MetabolismABSTRACT
[Objective]To investigate a minimally invasive,safe and effective method for treating the nonunion of tibia fracture with Bastianti external fixators.[Method]Bastianti external fixators were used to treat nonunion tibia fracture with axial force followed by functional exercises in early stage.[Result]All cases showed tibia fracture union in 4~12 months after operation.According to the criterion of treatment effects,35 cases were as excellent,7 as good,and 1 as poor.[Conclusion]It is a good method that nonunion of tibia fracture was treated with Bastianti external fixators for maintaining pain-free joint activity and joint functional recovery in early stage.