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Objective:To explore the role of corticotrophin releasing factor receptor 2 (CRFR2) in regulating pain sensitization and anxiety and its mechanism in chronic migraine mice.Methods:Forty-eight C57BL/6J mice were randomly divided into control group, model group, NBI35965 group and K41498 group ( n=12); chronic migraine models in the later 3 groups were established by intraperitoneally administrating 10 mg/kg nitroglycerin on the 1 st, 3 rd, 5 th, 7 th and 9 th d; mice in the NBI35965 group and K41498 group were injected with 100 nL NBI35965 or K41498 solution into the bilateral trigeminal nucleus caudalis on the 2 nd, 4 th, 6 th and 8 th d, and mice in the control group were injected with same volume of normal saline. Von frey fiber was used to detect the orbitofrontal mechanical pain threshold 2 h after intraperitoneal injection on the 1 st, 3 rd, 5 th, 7 th and 9 th d, and at 11 a.m. on the 10 th d. Elevated plus maze was used to detect the anxiety-like behaviors at 11 a.m. on the 11 th d. Western blotting was performed to detect the protein expressions of corticotrophin releasing factor (CRF), corticotrophin releasing factor receptor 1 (CRFR1), CRFR2 in the trigeminal nucleus caudalis. Real-time quantitative PCR (RT-qPCR) was used to detect the CRFR1 and CRFR2 mRNA expressions in the trigeminal nucleus caudalis. Immunofluorescent staining was used to detect the protein expressions of calcitonin gene-related peptide (CGRP), immediate-early gene c-fos, glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (Iba-1) in the trigeminal nucleus caudalis. Results:Compared with the control group, the model group, NBI35965 group and K41498 group had significantly decreased orbitofrontal mechanical pain thresholds 3, 5, 7, 9, and 10 d after intraperitoneal injection ( P<0.05); compared with model group, the K41498 group had significantly increased orbitofrontal mechanical pain thresholds 7, 9, and 10 d after intraperitoneal injection ( P<0.05). Compared with control group, the model group, NBI35965 group and K41498 group had significantly decreased entries and shorter time in opened arms ( P<0.05); compared with the model group, the K41498 group had significantly increased entries and shorter time in opened arms ( P<0.05). Compared with the control group, the model group, NBI35965 group and K41498 group had significantly higher CRF and CRFR2 protein expressions in the trigeminal nucleus caudalis ( P<0.05); compared with the model group, the K41498 group had statistically lower CRF protein expression in the trigeminal nucleus caudalis ( P<0.05). Compared with the control group, the model group, NBI35965 group and K41498 group had significantly higher CRFR2 mRNA expression in the trigeminal nucleus caudalis ( P<0.05). Compard with the control group, the model group, NBI35965 group and K41498 group had significantly increased CGRP, c-fos, Iba-1 and GFAP protein expressions in the trigeminal nucleus caudalis ( P<0.05); compared with the model group, the K41498 group had significantly decreased CGRP and c-fos protein expressions in the trigeminal nucleus caudalis ( P<0.05). Conclusion:CRFR2 can alter the orbitofrontal pain sensitization and anxiety-like behaviors in chronic migraine mice by regulating neuronal activation and CGRP release in the trigeminal nucleus caudalis.
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Aim To investigate the role of autophagy regulated by the AMPK/mTOR pathway in the prevention of oxygen-glucose deprivation/reperfusion injury ( OGD/R) in astrocytes using oxymatrine ( OMT ) . Methods The isolated and purified astrocytes ( AS) were randomly divided into control group ( CON group), OGD/R group and OGD/R + OMT group (0. 1, 0. 2, 0. 4 mmol · L
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@#Keratoplasty is routinely used to treat end-stage corneal diseases. However, immune-mediated graft rejection remains the major cause of surgical failure. Organ transplant rejection is often due to the directional migration and homing of inflammatory cells to lymphoid tissues and local inflammatory sites, which is regulated by various adhesion molecules and chemokines. Regulatory T cells play a key role in immune regulation and are essential for maintaining peripheral tolerance. Recent studies have revealed that regulatory T cells play important roles in preventing organ transplant rejection and the development of autoimmune diseases. This review will summarize the recent research on the induction of ocular immune privilege by regulatory T cells, with special focus on how regulatory T cells mediate tolerance in the eye and clinical potential of modulating these mechanisms during corneal transplantation.
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Objective@#To evaluate the effect of exogenous insulin on endoplasmic reticulum stress in myocardial tissues during insulin resistance in the rabbits undergoing cardiopulmonary bypass (CPB).@*Methods@#Forty healthy adult New Zealand white rabbits of both sexes, weighing 2.5-3.0 kg, were divided into 4 groups (n=10 each) using a random number table method: control group (group C), CPB group, CPB plus insulin group (group I) and CPB plus normal saline group (group NS). Group C received no treatment.An insulin resistance model was established in group CPB.Insulin was continuously infused (the infusion rate was adjusted according to the blood glucose) starting from establishing CPB to 1 day after operation in group I. The equal volume of normal saline was given starting from establishing CPB to 1 day after operation in group NS.Blood samples were collected from the left femoral artery, and myocardial tissues obtained before CPB (T1) and at 15, 30 and 60 min after aortic opening (T2-4). The level of blood glucose was determined using oxidase method, the level of glucagon was detected by the radioimmunoassay method, and the insulin resistance index was calculated.The expression of inositol-requiring protein-1α(IRE1α), XBP1 and caspase-12 was measured by Western blot.The expression of IRE1α, XBP1 and caspase-12 mRNA was measured by fluorescent quantitative real-time polymerase chain reaction.Cell apoptosis was detected by TUNEL.@*Results@#Compared with group C, blood glucose and glucagon levels at T2-4 and insulin resistance index at T4 were significantly increased, and the expression of inositol-requiring protein-1α(IRE1α), XBP1 and caspase-12 protein and mRNA was up-regulated at T4 in CPB, I and NS groups (P<0.05). Compared with group CPB, blood glucose and glucagon levels at T2-4 and insulin resistance index at T4 were significantly decreased, and the expression of IRE1α, XBP1 and caspase-12 protein and mRNA was down-regulated at T4 in group I (P<0.05).@*Conclusion@#Exogenous insulin significantly improves insulin resistance in myocardial tissues, and the mechanism is related to inhibiting endoplasmic reticulum stress and reducing cell apoptosis in the rabbits undergoing CPB.
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Objective To evaluate the effect of exogenous insulin on endoplasmic reticulum stress in myocardial tissues during insulin resistance in the rabbits undergoing cardiopulmonary bypass (CPB).Methods Forty healthy adult New Zealand white rabbits of both sexes,weighing 2.5-3.0 kg,were divided into 4 groups (n =10 each) using a random number table method:control group (group C),CPB group,CPB plus insulin group (group Ⅰ) and CPB plus normal saline group (group NS).Group C received no treatment.An insulin resistance model was established in group CPB.Insulin was continuously infused (the infusion rate was adjusted according to the blood glucose) starting from establishing CPB to 1 day after operation in group Ⅰ.The equal volume of normal saline was given starting from establishing CPB to 1 day after operation in group NS.Blood samples were collected from the left femoral artery,and myocardial tissues obtained before CPB (T1) and at 15,30 and 60 min after aortic opening (T2-4).The level of blood glucose was determined using oxidase method,the level of glucagon was detected by the radioimmunoassay method,and the insulin resistance index was calculated.The expression of inositol-requiring protein-1α(IRE1α),XBP1 and caspase-12 was measured by Western blot.The expression of IRE1α,XBP1 and caspase-12 mRNA was measured by fluorescent quantitative real-time polymerase chain reaction.Cell apoptosis was detected by TUNEL.Results Compared with group C,blood glucose and glucagon levels at T2-4 and insulin resistance index at T4 were significantly increased,and the expression of inositol-requiring protein-1α (IRE1α),XBP1 and caspase-12 protein and mRNA was up-regulated at T4 in CPB,I and NS groups (P<0.05).Compared with group CPB,blood glucose and glucagon levels at T2-4 and insulin resistance index at T4 were significantly decreased,and the expression of IRE1α,XBP1 and caspase-12 protein and mRNA was down-regulated at T4 in group Ⅰ (P<0.05).Conclusion Exogenous insulin significantly improves insulin resistance in myocardial tissues,and the mechanism is related to inhibiting endoplasmic reticulum stress and reducing cell apoptosis in the rabbits undergoing CPB.
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Objective To explore the impacts of natural ovulation cycles and stimulation cycles on the outcome of intrauterine insemination (IUI) in order to improve the clinical effects of IUI. Methods 176 women received 384 stimulation cycles. According to different ovulation stimulation protocols , the women were divided into six groups including natural cycle (NC) group, clomiphene citrate (CC) group, letrozole (LE) group;human menopausal gonadotrophin (HMG ) group, CC + HMG group, and LE + HMG group. The pregnancy rate between nature cycles and ovarian hyperstimulation cycles was compared. Results The pregnancy rate was 9.33%in the nature cycle group and 13.27% in the stimulation cycle group, with a significant difference (P 0.05). Conclusions Use of ovulation-induction medications is one of the important factors affecting the pregnancy rate of intrauterine insemination. There are no differences in the outcome of IUI among different ovulation stimulation protocols.
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Background Recent researches show that oxidative stress is involved in the progress of keratoconus.Nuclear factor-E2-related factor 2-antioxidant response element (Nrf2-ARE) pathway plays a critical role in the defense against oxidative stress,but its function in keratoconus is unclear.Objective To investigate the differences of Nrf2-ARE signaling activation and matrix degenerating enzymes between keratoconus and normal corneal stromal cells.Methods Corneal stromal cells were isolated from keratoconus and normal cornea by using dispase and collagenase digestion.The cells were treated with hydrogen peroxide (H2O2) to mimic in vivo oxidative stress condition.Reactive oxygen species (ROS) production was measured by fluorescence substrate DCHF-DA incubation.Nrf2 level and the expression of Nrf2-ARE downstream antioxidant genes were analyzed by Western blot and real-time quantitative-PCR(RT-qPCR).The activity of matrix degenerating enzymes,including urokinase-type plasminogen activator (uPA)-uPA receptor (uPAR) system and matrix metalloproteinase-2 (MMP-2) were assessed by Western blot and gelatin zymography respectively.Results In normal culture,keratoconus corneal stromal cells assumed increased basal ROS and Nrf2 level when compared with normal cells(t =18.155,P<0.01).However,after H2O2 treatment,the keratoconus corneal stromal cells showed increased ROS production,while decreased Nrf2 translocation and no significant difference in expression levels of Nrf2-ARE downstream antioxidant genes (Nrf2:t =62.123,P< 0.01 ; (nicotinamide adenine dinucleotide phosphate quinine oxidoreductase-1 [NQO-1]:t =2.209,P =0.092 ; hemo oxygenase-1 [HO-1]:t =0.293,P =0.784 ; superoxide dismutase [SOD2]:t =0.749,P =0.495).The contents of uPA-uPAR and the activity of MMP-2 also showed a higher level in keratoconus corneal stromal cells than normal cells,with significant differences between them (t =19.164,15.458,4.818,all at P<0.01).Conclusions The defect of Nrf2-ARE signaling activation exists in the keratoconus corneal stromal cells,and correlats with the abnormal expression level of stromal degeneration enzymes,which suggests that the defect of Nrf2-ARE signaling activation may be involved in the progression of keratoconus.
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<p><b>OBJECTIVE</b>To investigate the know gene types of main wild type measles virus strains and take measures to control measles in Jilin Province.</p><p><b>METHODS</b>Genetic characterization of 9 measles viruses isolated from 72 throat swabs or urine specimens of measles patients using CDW(150) cells line was studied in Jilin Province in 2005.</p><p><b>RESULTS</b>Sequence analysis of 450 nucleotides of COOH-terminal of nucleoprotein (N) genes of 9 isolates indicated that all were members of H(1) genotype, in which there are 7 strains of H1a and 2 strains of H1b, the H1a subgroup differed from H1b by 2.0% approximately 3.5% at the nucleotide level in the COOH-terminal of the N gene.</p><p><b>CONCLUSIONS</b>The H(1) genotype of wild-type measles viruses should be the main epidemic strain and main pathogen that caused measles outbreaks and sporadic cases in Jilin Province.</p>
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Humanos , China , Genes Reporteros , Genes Virales , Genotipo , Sarampión , Epidemiología , Virología , Virus del Sarampión , Genética , Datos de Secuencia Molecular , Proteínas Estructurales Virales , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To study the effect of rhTNF-alpha on human sperm mitochondrial function and motility in vitro.</p><p><b>METHODS</b>Fifty-six semen samples collected by masturbation were analyzed according to WHO protocols. Semen samples from 40 healthy men were prepared using Percoll centrifugation. Sperm suspension was diluted to a concentration of 10 x 10(6)/ml in Ham's F10 medium. Sperm samples were incubated with rhTNF-alpha solution (final concentration 0.03 microg/L, 0.06 microg/L, 0.09 microg/L and 0.27 microg/L, respectively) for 0.5 h, 1 h, 2 h, 3 h and 4 h at 37 degrees C in 5% CO2, and comparative studies were made with a control group. Ten microl sperm samples were examined with CASA technique, 250 microl stained in the presence of 10 microg/ml Rh123 and PI, and mitochondrial function analyzed by flow cytometry.</p><p><b>RESULTS</b>Significant differences were found between the experimental groups (final concentration 0.06 microg/L, 0.09 microg/L and 0.27 microg/L) and the control group in viability, straight line velocity, curvilinear velocity, average path velocity, progressive motility of human sperm and the number of spermatozoa with normal mitochondrial function (P < 0.01) except the final concentration 0.03 microg/L group (P > 0.05). Motility of human sperm lowered with the increase of rhTNF-alpha concentration and incubation time, and r values were 0.675, 0.691, 0.762, 0.693, 0.724 and 0.571, 0.594, 0.752, 0.791, 0.816, respectively (P < 0.01). The number of spermatozoa with normal mitochondrial function decreased with the increased rhTNF-alpha concentration and incubation time, and r values were 0.615, 0.643, 0.752, 0.691, 0.754 and 0.532, 0.567, 0.782, 0.692, 0.854, respectively (P < 0.01).</p><p><b>CONCLUSION</b>rhTNF-alpha can reduce human sperm motility function in vitro, possibly by interfering with human sperm mitochondrial function.</p>
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Adulto , Humanos , Masculino , Relación Dosis-Respuesta a Droga , Mitocondrias , Fisiología , Proteínas Recombinantes , Farmacología , Motilidad Espermática , Factor de Necrosis Tumoral alfa , FarmacologíaRESUMEN
<p><b>OBJECTIVES</b>To investigate the possibility and efficacy of vasoligation and deferent vein ligation in treating and preventing benign prostatic hyperplasia(BPH).</p><p><b>METHODS</b>40 male pooches were divided into A, B, C and D groups randomly. Each group has 10 pooches group A and B were made into the model of BPH. Two years later, the pooches in group A and C accepted the operation of vasoligation and deferent vein ligation on both sides. Group B and D only accepted the operation of dissection and relieving its deferent duct. Then continuing to feed the 40 pooches after the operation, they were killed another two years later. After taking out their prostates and calculating their volume and weight, the sections of the prostates were checked by microscope to observe their histology and pathology changes.</p><p><b>RESULTS</b>There were significant differences of weight and volume as well as the changes of histology and pathology between group A and group C (P < 0.01). It was the same with group B and D (hyperplasia changes).</p><p><b>CONCLUSIONS</b>Vasoligation and deferent vein ligation before benign prostatic hyperplasia at pooches grow-up stage can reduce the extent of BPH at old age, and treatment after benign prostatic hyperplasia can make prostatic tissue appear different degrees of atrophy.</p>