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Aim To investigate whether and how Huy- izhenbao tablets regulated osteoarthritis pain.Methods We transected the meniscotibial ligament of mice, which caused osteoarthritis by destabilizing the medial meniscus ( DMM).Different doses of Huyizhenbao tablets (12.5,25,50 mg • kg-1) were administered intragastrically.Dynamic and static mechanical allo- dynia were measured.The spinal cord slices were pre¬pared to record miniature excitatory postsynaptic cur-rents (niEPSCs) and miniature inhibitory postsynaptic currents (mlPSCs) by using patch clamp electrophysi¬ological recordings.The phosphorylation of NMDA re¬ceptor ( N-methyl-D-aspartate receptors) (rluNl sub- unit at S897 residue ( pS897-GluNl ) was observed by immunohistochemistry.Results Huyizhenbao tablets dose-dependently attenuated the dynamic and static mechanical allodynia induced by DMM, reduced the frequency of niEPSCs and inhibited the pS897-GluNI level.Huyizhenbao tablets had no effects on mlPSCs.Conclusions Huyizhenbao tablets effectively alleviate osteoarthritis pain by blocking the presynaptic release of excitatory transmitter glutamate and inhibiting the phosphorylation of NMDA receptor in spinal cord dorsal horn.
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BACKGROUND@#Bile duct injury (BDI), which may occur during cholecystectomy procedures and living-donor liver transplantation, leads to life-altering complications and significantly increased mortality and morbidity. Tissue engineering, as an emerging method, has shown great potential to treat BDI. Here, we aimed to explore the application of small intestinal submucosa (SIS) matrix composites with bone marrow mesenchymal stem cells (BMSCs) to treat BDI in a rabbit model. @*METHODS@#Rabbit-derived BMSCs were used as seed cells. Porcine SIS was used as the support material. Five centimetres of the common bile duct was dissected, and 1/3–1/2 of the anterior wall diameter was transversely incised to construct the rabbit BDI model. Then, SIS materials without/with BMSCs were inserted into the common bile duct of the BDI rabbits. After 1, 2, 4, and 8 weeks of implantation, the common bile duct was removed. Haematoxylin and eosin (HE) staining was used to assess pathological alterations in the common bile duct, while immunohistochemical staining and western blotting were used to detect expression of the epithelial cell markers CK19 and E-cadherin. Scanning electron microscopy was used to evaluate BMSC growth. @*RESULTS@#Compared with BMSCs alone, SIS-attached BMSCs had increased growth. HE staining showed that the injured bile duct healed well and that the complex gradually degraded as the time from implantation increased. Immunohistochemical staining and western blotting showed that compared with the control group, the in vivo complex group had significantly elevated expression levels of CK19 and E-cadherin. @*CONCLUSION@#BMSC implantation into SIS could improve BDI in rabbits, which might have clinical value for BDI treatment.
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BACKGROUND@#Bile duct injury (BDI), which may occur during cholecystectomy procedures and living-donor liver transplantation, leads to life-altering complications and significantly increased mortality and morbidity. Tissue engineering, as an emerging method, has shown great potential to treat BDI. Here, we aimed to explore the application of small intestinal submucosa (SIS) matrix composites with bone marrow mesenchymal stem cells (BMSCs) to treat BDI in a rabbit model. @*METHODS@#Rabbit-derived BMSCs were used as seed cells. Porcine SIS was used as the support material. Five centimetres of the common bile duct was dissected, and 1/3–1/2 of the anterior wall diameter was transversely incised to construct the rabbit BDI model. Then, SIS materials without/with BMSCs were inserted into the common bile duct of the BDI rabbits. After 1, 2, 4, and 8 weeks of implantation, the common bile duct was removed. Haematoxylin and eosin (HE) staining was used to assess pathological alterations in the common bile duct, while immunohistochemical staining and western blotting were used to detect expression of the epithelial cell markers CK19 and E-cadherin. Scanning electron microscopy was used to evaluate BMSC growth. @*RESULTS@#Compared with BMSCs alone, SIS-attached BMSCs had increased growth. HE staining showed that the injured bile duct healed well and that the complex gradually degraded as the time from implantation increased. Immunohistochemical staining and western blotting showed that compared with the control group, the in vivo complex group had significantly elevated expression levels of CK19 and E-cadherin. @*CONCLUSION@#BMSC implantation into SIS could improve BDI in rabbits, which might have clinical value for BDI treatment.
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Graves' disease(GD) is the most common cause of endogenous thyrotoxicosis, and current therapeutic options for GD include antithyroid drugs(ATD), radioactive iodine and thyroidectomy. Refractory hyperthyroidism is a rare condition in which hy¬perthyroidism fails to respond to the conventional therapy of anti-thyroidal med¬ications. Enlarged goiter size, high titers of thyrotropin receptor antibodies(TRAb) at diagnosis and/or at the end of the treatment, TSH suppression after drug withdrawal and compliance to the treatment have all been proposed as risk factors that predict the relapse of GD. we propose that a patient-centered professionalism should be consistently followed, and that refractory hyperthyroidism because of unsuitable therapeutic strategies should be strictly avoided in the clinical practice. For the patients with refractory hyperthyroidism, radical treatment is recommended, however,continued long-term low-dose ATD can be considered.
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<p><b>OBJECTIVE</b>To explore the effect of diallyl disulfide (DADS) on hippocampal synapses and learning and memory abilities in a mouse model of A1zheimer's disease (AD).</p><p><b>METHODS</b>Mouse models of AD established by agglutinated Aβ1-42 injection in the lateral cerebral ventricle were randomized into 4 groups and treated with DADS at the daily doses of 0, 10, 50 and 100 mg/kg by gavage for 30 consecutive days. The learning and memory abilities of the mice were assessed with Morris water maze test; the structures of the dendritic spines and synapses in CA1 region of the hippocampus were observed under transmission electron microscope with silver staining; PSD95 and SYP protein and mRNA expressions in the hippocampus were detected with Western blotting and RT-PCR.</p><p><b>RESULTS</b>Compared with the AD model mice, the mice treated with 50 and 100 mg/kg DADS showed enhanced learning and memory abilities in Morris water maze test. The dendritic spines and synapses in CA1 region of the hippocampus increased obviously and hippocampal expressions of PSD95 and SYP were enhanced in mice treated with 50 and 100 mg/kg DADS.</p><p><b>CONCLUSION</b>DADS at the daily doses of 50 and 100 mg/kg can improve the learning and memory abilities and increase the number of dendritic spines and synapses in the hippocampus in mouse models of AD.</p>
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Animais , Masculino , Camundongos , Compostos Alílicos , Farmacologia , Doença de Alzheimer , Tratamento Farmacológico , Região CA1 Hipocampal , Modelos Animais de Doenças , Dissulfetos , Farmacologia , Aprendizagem , Memória , SinapsesRESUMO
<p><b>OBJECTIVE</b>To analyze the clinical outcomes of intracytoplasmic sperm injection (ICSI) with spermatozoa from different sources.</p><p><b>METHODS</b>We retrospectively studied the rates of fertilization, clinical pregnancy, implantation, abortion, ectopic pregnancy and delivery in 682 patients treated by ICSI, who were divided according to the sperm sources into an ejaculated sperm group (ES, n = 598), a percutaneous epididymal sperm aspiration (PESA, n = 58) and a testicular sperm extraction (TSE, n = 26).</p><p><b>RESULTS</b>The fertilization rate was significantly lower in the TSE than in the ES and PESA groups (81.06% vs 87.95% and 87.82%, P < 0.05). But no statistically significant differences were observed among the ES, PESA and TSE groups in the rates of clinical pregnancy (39.46%, 48.28% and 34.62%), implantation (19.80%, 23.80% and 18.34%), abortion (13.13%, 17.86% and 11.11%), ectopic pregnancy (5.51%, 7.14% and 11.11%) and delivery (32.11%, 36.21% and 26.92%) (P > 0.05).</p><p><b>CONCLUSION</b>Although TSE-obtained sperm affect the rate of fertilization, those obtained by TSE and PESA do not obviously influence the outcome of clinical pregnancy.</p>
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Adulto , Feminino , Humanos , Masculino , Gravidez , Resultado da Gravidez , Estudos Retrospectivos , Injeções de Esperma Intracitoplásmicas , Métodos , Recuperação Espermática , EspermatozoidesRESUMO
<p><b>OBJECTIVE</b>To investigate the chromosome segregation patterns in patients with chromosome balanced translocation in spermatogenesis and to find out different proportions of abnormal sperm generated by different patterns.</p><p><b>METHODS</b>Triple color fluorescence in-situ hybridization (FISH) was used to determine the correlative chromosome constitution of spermatozoa from 4 patients with chromosome anomalies, Case 1 with 46, XY, t(9;11)(q22;q21), Case 2 with 46, XY, t(11;22) (q23;q11), Case 3 with 45, XY, t(13q;15q) and Case 4 with 45, XY, t(13q;14q). The numbers of spermatozoa generated by different chromosome segregation patterns were counted by FISH, and normal sperm from the males with normal chromosome was used as control.</p><p><b>RESULTS</b>The rates of abnormal sperm in the 4 cases were 50.86%, 58.33%, 13.00% and 22.82% respectively, evidently higher than their corresponding controls (0.85%, 1.63%, 1.60% and 1.37%).</p><p><b>CONCLUSION</b>To detect sperm chromosome in patients with chromosome balanced translocation by FISH can provide valuable information for predicting the risk of chromosome anomalies in the embryo preimplantation genetic diagnosis.</p>
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Adulto , Humanos , Masculino , Transtornos Cromossômicos , Diagnóstico , Genética , Hibridização in Situ Fluorescente , Métodos , Cariotipagem , Espermatozoides , Biologia Celular , Metabolismo , Translocação GenéticaRESUMO
<p><b>OBJECTIVE</b>To evaluate the influence of big Y chromosome on the outcomes of in vitro fertilization and embryo transfer.</p><p><b>METHODS</b>Data of 127 cycles of IVF/ICSI-ET, performed in our Reproductive Medicine Center from March 2001 to June 2003 were analyzed. The patients were divided into two groups according to the length of chromosome: Group A, 56 cycles with big Y chromosome</p><p><b>RESULTS</b>No significant difference was observed in the quality of embryos and in the and Group B, 71 cycles with normal karyotype. rates of fertilization, cleavage, clinical pregnancy, implantation, miscarriage, ectopic pregnancy, dead infant delivery, malformation,</p><p><b>CONCLUSION</b>Big Y chromosome has no significant influence on the baby boy delivery and baby girl delivery between the two groups. development of embryos and the outcome of pregnancy.</p>
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Feminino , Humanos , Masculino , Gravidez , Cromossomos Humanos Y , Implantação do Embrião , Infertilidade Masculina , Genética , Terapêutica , Taxa de Gravidez , Estudos Prospectivos , Injeções de Esperma IntracitoplásmicasRESUMO
Our previous studies have shown that long-term potentiation (LTP) of C-fiber-evoked field potentials in the spinal dorsal horn is NMDA receptor dependent. It is known that elevation of Ca(2+) in the postsynaptic neurons through NMDA receptor channels during high-frequency stimulation of the afferent fibers is crucial for LTP induction, but how this leads to a prolonged potentiation of synaptic transmission in the spinal dorsal horn is not clear. In the hippocampus, a rise of Ca(2+) activates calcium/calmodulin-dependent protein kinase II (CaMK II) through autophosphorylation. Once this occurs, the kinase remains active, even when Ca(2+) concentration returns to baseline level. Phosphorylated CaMK II potentiates synaptic transmission by enhancement of AMPA receptor channel function via phosphorylation of GluR1 subunit of the receptor and the addition of AMPA receptors to synapses. Up to now, the role of CaMK II in the induction and maintenance of LTP of the C-fiber-evoked field potentials in spinal dorsal horn has not been evaluated. In the present study, we examined the expression of CaMK II and phospho-CaMK II in the lumbar segments (L4-L6) of the rat spinal dorsal horn at 30 min and 3 h after the establishment of LTP induced by tetanic electrical stimulation of the sciatic nerve (40 V, 0.5 ms pulses at 100 Hz for 1 s repeated four times at 10 s intervals) by using Western blot and electrophysiological techniques. To determine the role of the phospho-CaMK II in the induction and maintenance of the spinal LTP, a selective CaMK II inhibitor KN-93 (100 micromol/L) was applied directly onto the spinal cord at the recording segments before and after LTP induction. We found that (1) the protein level of phospho-CaMKII increased at both 30 min and 3 h after LTP induction, while the total protein level of CaMK II increased at 3 h but not at 30 min after LTP induction. (2) Spinal application of KN-93 at 30 min prior to the tetanus blocked both LTP induction and the increase in phospho-CaMK II. (3) 30 min after LTP induction, spinal application of KN-93 depressed LTP and the level of phospho-CaMK II (n=3). (4) Spinal application of KN-93 at 3 h after LTP, however, affected neither the amplitude of the spinal LTP nor the level of phospho-CaMK II in the spinal dorsal horn. These results suggest that activation of CaMK II is probably crucial for the induction and the early-phase maintenance of LTP of C-fiber-evoked field potentials in the spinal dorsal horn.
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Animais , Masculino , Ratos , Potenciais Evocados , Potenciação de Longa Duração , Fisiologia , Fibras Nervosas Amielínicas , Fisiologia , Vias Neurais , Fisiologia , Fosfoproteínas Fosfatases , Metabolismo , Fosforilação , Células do Corno Posterior , Fisiologia , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato , Medula Espinal , FisiologiaRESUMO
Nerve injury produces a long lasting neuropathic pain, manifested as allodynia, a decrease in pain threshold and hyperalgesia, an increase in response to noxious stimuli. The mechanism underlying the lasting abnormal pain is not well understood. Our previous works have shown that electrical tetanic stimulation of the sciatic nerve induces long-term potentiation (LTP) of C-fiber evoked field potentials in the spinal dorsal horn, which is considered as a synaptic model of pathological pain. In the present study we tested if nerve injury, which is proved to produce neuropathic pain, induced the spinal LTP in intact rats. C-fiber evoked field potentials in spinal dorsal horn produced by electrical stimulation (10-20 V, 0.5 ms, 1/min) of the sciatic nerve were recorded. For induction of LTP of C-fiber evoked field potentials, three types of noxious stimuli were applied. (1) Electrical tetanic stimulation (40 V, 0.5 ms pulses at 100 Hz for 1 s repeated four times at 10 s intervals). (2) Transection of the sciatic nerve at 4-5 mm distal to the stimulation electrode. (3) Crushing the sciatic nerve with a forceps four times at 4-5 mm distal to stimulation electrode (from distal to proximal with 1 mm spacing at 10 s intervals), which simulated electrical tetanic stimulation. Acute nerve injury was made by either transection of the sciatic nerve at the distal to the stimulating electrode or crushing the sciatic nerve. We found that nerve injury by cutting or crushing the sciatic nerve produced LTP of C-fiber evoked field potentials lasting until the end of the experiments (3-9 h), and that pretreatment of the sciatic nerve with lidocaine 10 min prior to the nerve transectoin completely blocked LTP induced by nerve transection. The nerve transection-induced LTP was blocked by NMDA receptor antagonist AP5. LTP produced by nerve transection could not be further potentiated by electrical tetanic stimulation, while LTP induced by single electrical tetanic stimulation could be further potentiated by transection of the sciatic nerve. However, when LTP was saturated by several times of electrical tetanic stimulation, nerve transection did not affect the spinal LTP. We conclude that acute nerve injury induces LTP of C-fiber evoked field potentials in intact animals and that nerve transection is more powerful than electrical tetanic stimulation for induction of the spinal LTP. The results further support the notion that LTP of C-fiber evoked field potentials may underlie neuropathic pain.