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Objective To investigate the expression of oxytocin ( OXT ) and oxytocin receptor (OXTR) in the prefrontal cortex of postpartum depression (PPD) rats induced by restraint stress during pregnancy and to observe the antidepressant effect of oxytocin and its analogue capitoxin and its mechanism. Methods Twenty-four adult female SD rats of SPF grade were randomly divided into control group,PPD +saline group,PPD + oxytocin group and PPD + captopril group with 6 rats in each group. Rats were subjec-ted to restraint stress for 2 hours every day on the 8th to 21st day of pregnancy to establish PPD model. While the rats in control group were not given any treatment. Rats in PPD + saline,PPD + oxytocin and PPD +captopril were injected bilaterally into prefrontal cortex (PFC) at 10 days postpartum (1 μl/side),oxytocin (30 ng/side) and captopril (45 ng/side) respectively once a day for 5 days. The depressive behaviors of rats were detected by sugar-water preference experiment. Rats were sacrificed 18 days after delivery. The ex-pression of OXT was detected by ELISA method,OXTR by Western blot,Iba-1 by immunofluorescence,and IL-1β,IL-6 and TNF-α by qRT-PCR. Results (1) The sucrose consumption of the PPD + saline group ((67. 1±10. 4)%) was significantly lower than that of the control group((92. 6± 3. 9)%,t=-5. 31,P<0. 01). (2) The expression of oxytocin in prefrontal cortex in PPD group was significantly lower than that in control group ((0. 03±0. 01) ng/mg) vs (0. 08 +0. 05) ng/mg,t=-2. 67,P<0. 05). However,there was no significant difference in the expression of oxytocin receptor between PPD group and control group ((0. 90 ±0. 06) vs (0. 90±0. 05),t=0. 709,P=0. 517). (3) The sucrose consumption of PPD+saline group de-creased than that of control group((65. 6±16. 9)% vs (91. 5±3. 5)%,t=3. 35,P<0. 001). Compared with PPD+saline group,the sucrose consumption of PPD+oxytocin group ((81. 8±8. 4)%) and PPD+carbetocin group ((78. 4±9. 4)%) increased(t=1. 98,1. 68,both P<0. 05). (4) The expression of Iba-1 in the pre-frontal lobe of PPD + saline group was higher than that of control group ((1. 15±0. 05) vs (1. 04 +0. 06), t=3. 50,P<0. 01). Compared with PPD + saline group,the expression of Iba-1 in PPD + oxytocin group (1. 03±0. 06) and in PPD + captopril group (1. 00±0. 02) were lower (t=-3. 50,-6. 55,both P<0. 01). (5) The expression of inflammatory factors IL-1β mRNA (1. 0±0. 1),IL-6 mRNA (1. 1±0. 1) and TNF-α mRNA (1. 7±0. 4) in the prefrontal cortex of rats in the PPD group were higher than that in the control group (IL-1β mRNA (0. 7± 0. 3),IL-6 mRNA (0. 9± 0. 1),TNF-α mRNA ( 1. 1± 0. 3),t=1. 92,3. 19, 2. 43 respectively,all P<0. 05). The expression of inflammatory factors IL-1β,IL-6 and TNF-α mRNA of the PPD+oxytocin group(IL-1β mRNA (0. 6±0. 1),IL-6 mRNA (0. 9±0. 1),TNF-α mRNA (1. 2±0. 4) )and the PPD+carbetocin group ( IL-1β mRNA ( 0. 7± 0. 1),IL-6 mRNA ( 0. 9 ± 0. 1),TNF-α mRNA ( 1. 0 ± 0. 2))in the prefrontal cortex were lower than that in the PPD group(t=-3. 17,-2. 78,-1. 84,t=-2. 76,-2. 40,-2. 94 respectively,all P<0. 05). Conclusion Oxytocin and capitoxin injected into prefrontal cortex can effectively improve depression-like behaviors in PPD model rats. Activation of microglia and decrease of inflammatory factors in prefrontal cortex may be the potential antidepressant mechanism.
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BACKGROUND: Pseudoachondroplasia is rare heredigary osteopathy, usually accompanied by deformity of the lower limbs, short fingers, and ligamentous laxity, so it is a difficulty in total knee arthroplasty.OBJECTIVE: To evaluate the curative effectiveness of bilateral total knee arthroplasties for a patient with pseudoachondroplasia using three-dimensional (3D) printing technology.METHODS: A retrospective study was done in a case of pseudoachondroplasia undergoing bilateral total knee arthroplasties using 3D printing technology. X-ray examination was conducted to observe the prosthesis location after arthroplasties, and the Knee Society Score outcome measures were used to evaluate the functional outcome.RESULTS AND CONCLUSION: (1) After arthroplasties, X-rays showed good line of force of the lower limbs and prosthesis location, and 1 year later, showed no radiolucent lines in tibia. (2) The right Knee Society Score improved to 94 points, the functional scoring improved to 90 points, and the postoperative flexion was 95°; the left Knee Society Score improved to 93 points, the functional scoring improved to 90 points, and the postoperative flexion was 90°. All patients were very satisfied with the treatment outcomes. (3) These results indicate that the individualized treatment scheme designed using 3D printing technology can reduce the surgical difficulty and trauma with high accuracy, and promote the functional recovery of the knee. Additionally, it obtains good clinical efficacy and patient's satisfaction in orthopaedic surgeries.
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@#Objective To construct dual-luciferase reporter plasmids containing the wild type and mutant rat extracellular signal-regulat-ed kinase 1 (ERK1) gene 3' untranslated regions (UTR) which were used to detect rno-miR-15b-5p's putative target gene. Methods The rat ERK1 gene 3' UTR fragment was amplified by polymerase chain reaction (PCR) from PC12 cell cDNA and cloned into pmiR-RB-ReportTM vector. The mutant rat ERK1 gene 3' UTR fragment was obtained by overlap PCR and inserted into pmiR-RB-ReportTM vector. Successful wild type and mutant recombinant plasmids were confirmed by DNA sequencing. PC12 cells were co-transfected with rno-miR-15b-5p mim-ic and pmiR-ERK13' UTR or pmiR-ERK1-mut 3' UTR and then analyzed by dual-luciferase reporter assay system. The achieved mutation sequence of the target site TGCTGCT was mutated to CGAACGT and GTACACG, respectively. Results The wild-type reporter vector pmiR-ERK13' UTR and the mutant reporter vector pmiR-ERK1-mut 3' UTR were successfully constructed. The rno-miR-15b-5p mimic de-creased the activity of pmiR-ERK13' UTR plasmid (P<0.001) but did not decrease the activity of pmiR-ERK1-mut 3' UTR plasmid. Conclu-sion The recombinant pmiR-ERK13' UTR and pmiR-ERK1-mut 3' UTR plasmids were constructed successfully, and luciferase activities demonstrated that the 3' UTR of ERK1 gene might be a potential target of rno-miR-15b-5p.
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Objective To explore the curative effect of deep-brain magnetic stimulation (DMS) on learned helplessness behavior in the chronic restrained stress (CRS) rat model. Methods Twenty-nine Sprague-Dawley rats were randomly divided into control group (n=8) and CRS group (n=21). CRS group was exerted chronic restrained stress, while the control group did not receive any stress, for three weeks. Then learned helplessness behavior was tested using Forced Swimming Test (FST) and the hopeless rats of the CRS group were divided ran-domly into sham group (n=6), DMS group (n=8) and citalopram group (n=7), that received corresponding treatment respectively. They were evaluated with FST again after one-week treatment. Results The immobile time in FST was longer in CRS group than in the control group after three-week stress (F=11.260, P=0.002). After one-week treatment, no significant improvement was found in the citalopram group (F=1.565, P=0.235), however, the immobile time in DMS group decreased (F=6.277, P=0.025), and was shorter than that in the sham group (F=5.560, P=0.036). Conclusion CRS could result in learned helplessness behavior, which could be alleviated with one-week DMS.
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Objective: To evaluate the effectiveness of total knee arthroplasty (TKA) using three-dimensional (3D) printing technology for knee osteoarthritis (KOA) accompanied with extra-articular deformity.
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Objective To explore the effect of galanin ( GAL) on locus coeruleus ( LC) neurons from neonatal rat and mechanism with its receptor GalR and potassium channel.Methods Brain slices from neonatal rats were prepared and the resting membrane potential and spontaneous action potential of LC neurons were recorded with whole cell patch-clamp configuration.GAL, AR-M1896 and potassium channel blockers were bath applied with different concentration.Results Bath application GAL induced hyperpolarization and inhibited firing rate of LC neurons.However, AR-M1896 ( a selective GalR2 agonist) did not induce significant effect on LC neurons, only at very high concentration(1μM) it induced slight hyperpolarization and reduced firing rate.The inhibitory effect of GAL was partially blocked by TEA ( an antagonist of voltage-dependent potassium channel) and significantly blocked by BaCl2(an antagonist of inward-rectifying potassium channels), while other potassium channels blockers such as Glybenclamide(ATP-sensitive potassium channel blocker),Charybdotoxin(large-conductance Ca2 +-activated K + channels blocker),Apamin(small-conductance Ca2 +-activated K +channels blocker) failed to block it.Conclusion GAL inhibits LC neurons from neonatal rats, mainly through GalR1.TEA-sensitive potassium channels and inward-rectifying potassium channels, but not ATP-sensitive potassium channel and calcium-activated potassium channel, are involved in this inhibition.
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Objective To explore the effect of intracerebroventricular injection of AR-M1896, a galanin receptor 2 agonist, on depres-sion-like behavior in rat chronic mild stress (CMS) model. Methods 48 Sprague-Dawley rats were randomly assigned into control group, CMS group, CMS artificial cerebrospinal fluid (aCSF) group and CMS AR-M1896 group equally. The control group received no interven-tion, and the other groups were established chronic mild stress model. After six-week of stress, forced swim test and sucrose preference test were conducted to identify the CMS rats. AR-M1896 or aCSF was injected into the lateral ventricle of CMS AR-M1896 group and CMS aC-SF group, respectively. The immobility time and climbing time in the forced swim test were analysed, and the sucrose consumption percent-age in the sucrose preference test was measured. Results The immobility time decreased (F=11.998, P<0.01), climbing time increased (F=8.268, P<0.05), and the sucrose consumption percentage increased (F=10.352, P<0.01) in CMS AR-M1896 group, compared with CMS aC-SF group. Conclusion Intracerebroventricular administration of galanin receptor 2 agonist AR-M1896 is effective on depression in CMS model rats.
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Depression is the most common mental disorder in epilepsy, which means that they maybe have some common pathogene-sis. This paper discussed the biological relation mechanism, such as neurotransmitter, neuropeptide and its receptor, glial cell, immune-medi-ators, nerve signal transduction pathway, synaptic plasticity and nerve regeneration.
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Objective To study the expression and role of galanin in the hypothalamus of rat with the drug-induced hyperprolactinemia.Methods Hyperprolactinemia was induced by daily intraperitoneal injections 50 mg/kg sulpiride solution.The protein in the hypothalamus of rat was extracted to determine the expression levels of galanin with Western Blot.The expression and colocalization of galanin and dopamine in model and control group were observed with immunofluorescence.Results The model group showed a significant increase of serum prolactin (PRL) level and a significant decrease of serum estradiol (E2) level,as compared to the control group ((15.74±2.49) ng/ml vs (10.25±1.29) ng/ml and (4.24±0.69)pg/ml vs (9.56±3.25) pg/ml,respectively,P＜0.05).Both the expression level of galanin and the number of neurons coexisting with galanin and dopamine were decreased in the hypothalamus of the hyperprolactinemia rat compared with the control group.Western Blot revealed that,compared to the control group,the sulpiride model group had a significant increase of galanin but not TH (0.405±0.112 vs 0.985±0.158,P＜0.05 and 0.871 ± 0.046 vs 0.890±0.054,P＞ 0.05,respectively).Conclusion Galanin expression level has decrease in the hypothalamus of the hyperprolactinemia rat,which contributes to the reduction of dopaminergic neurons.
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Objective To study the protective effect of galanin on 2,3-Dimethoxy-1,4-naphthoquinone (DMNQ)-induced oxidative stress and cell damage in HEK-293A cells and its possible mechanisms.Methods The expression of galanin and its three receptors (GalR1-3) in HEK-293A were determined with RTPCR technique.The cultured HEK-293A cells were divided into four groups:Control,DMNQ,DMNQ + GAL and DMNQ + AR-M1896 and the cell viability was measured with CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS).Results The RT-PCR data revealed the presence of galanin and its receptors in HEK-293A cells.The expression level was in the order of GalR2 =Galanin ＞ GalR3 ＞ GalR1.DMNQ caused oxidative stress and cell damage in HEK-293A cells with a dose-dependent manner with an IC50 =13.4 μM.Application of galanin reduced the DMNQ-induced cellular toxicity in HEK-293A,which increased cell viability by 24.4％ and 18.8％ in 1 μM and 100 nM,respectively.AR-M1896,an agonist of GalR2 had a similar effect,increased cell viability by 8.7％ and 8.9％ in 1 μM and 100 nM,respectively.Conclusion These data suggest that galanin has a protective effect on DMNQ-induced oxidative stress and cell damage in HEK-293A cells,probably mediated by GalR2.
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Objective To explore the mechanisms of trafficking and signaling of serotonin 1A receptor(5-HT_(1A))and its spatiotemporal distribution in living cells.Methods The mouse 5-HT_(1A) gene amplified by RT-PCR was recombined into pEGFP-N1 vector and the EGFP coding sequence was located in-frame at the C-terminal end of the 5-HT_(1A) receptor.The 5-HT_(1A)-EGFP was transfected into neuron-like PC12 cells as well as HEK293.The transfected cells were visualized using confocal microscopy,the mobility of 5-HT_(1A)-EGFP was monitored by live measurements and fluorescence recovery after photobleaching.Results The 5-HT_(1A) gene was identitical with the published gene sequence NM_008308.4 and a 5-HT_(1A)-EGFP fusion construct was created.After stable transfection of the plasimd into a PC12 cell line and analysis with a confocal laser scanning microscopy,the EGFP-tagged 5-HT_(1A) was predominantly associated with the plasma membrane,but some intracellular vesicles in the perinuclear region also contained the fusion protein.The predominant localization of 5-HT_(1A)-EGFP at the plasma membrane was confirmed in transiently transfected HEK293 cells.Bleached fluorescence was partialy recovered in 100 seconds,indicating that the 5-HT_(1A)-EGFP was mobiled on the membrane.Conclusion Spatiotemporal distribution and mobility of 5-HT_(1A) tagged with EGFP can be monitored in the 5-HT_(1A)-EGFP stable PC12 cell line,which could be an excellent neuron-like experimental cell model for research of 5-HT_(1A) trafficking and signaling.
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Objective: To study the protective effect of glutamine on intestine in mice damaged by radiation. Methods: Mice were exposed to 137 Cs rays (dose of 4Gy) with the whole body. The protetive effects of glutamine on intestine radiational damages were studied. Result: After radiation, GSH contents, SOD activities, protein and DNA contents were decreased and MDA contents were increased in intestinal mucosa. Villus height was shortened, crypt depth was deepened, wall width and villus surface area were decreased. Glutamine supplement restrained the changes of indices above mentioned and made them close to normal levels. Conclusion: Glutamine supplement had the protective effect on radiation damage in mice.