Objective:To explore whether the muscarinic acetylcholine receptor M1(M1 AChR)regulate the expression of ionotropic glutamate receptor AMP A type subunit 1(GluA1)and ionotropic glutamate receptor AMP A type subunit 2(GluA2),thus participating in the effects of plateau hypoxia on learning and memory in rats.Methods:Rats were randomly divided into normoxic group(Control),plateau hypoxia group(Hypoxia)and plateau hypoxia+TAK-071(an agonist of M1AChR)group.Morris water maze was utilized to detect the learning and memory ability of rats.Immunohistochemistry was utilized to determine the distribution and expression levels of M1 AChR,GluA1,and GluA2 in hippocampal CA1 and CA3 areas of rats.Western Blot was utilized to determine the expression levels of M1 AChR,GluA1,p-GluA1(Ser845),GluA2 and p-GluA2(Ser880)in hippocampus of rats.RT-qPCR was utilized to determine the mRNA expression levels of M1 AChR,GluA1,GluA2,ionotropic glutamate receptor NMDA type subunit 2A(NR2A),tau and amyloid β-protein(Aβ)in hippocampus of rats.Results:Plateau hypoxia could significantly increase the escape latency of rats in the spatial navigation test and shorten the stay time of rats in the target quadrant in the spatial probe test,while TAK-071 treatment could significantly improve the memory status of rats damaged by plateau hypoxia in the spatial probe test.There was a good correlation between the expression of GluA1,GluA2 and M1 AChR in CA1 and CA3 regions of hippocampus.In hippocampus,the expressions of GluA1 and GluA2 were synchro-nized with their corresponding phosphorylated proteins,and the correlativity between GluA1 and GluA2 and M1 AChR changed due to plateau hypoxia was reversed after TAK-071 treatment.TAK-071 could significantly reduce(P＜0.05)the expression levels of M1 AChR,GluA1,GluA2,NR2A,tau and Aβ mRNA increased due to plateau hypoxia.Conclusion:Activating M1 AChR can improve the memory impairment of rats caused by plateau hypoxia by regulating the expression of GluA1,GluA2 and their phosphorylated proteins.

Objective:To explore whether the muscarinic acetylcholine receptor M1(M1 AChR)regulate the expression of ionotropic glutamate receptor AMP A type subunit 1(GluA1)and ionotropic glutamate receptor AMP A type subunit 2(GluA2),thus participating in the effects of plateau hypoxia on learning and memory in rats.Methods:Rats were randomly divided into normoxic group(Control),plateau hypoxia group(Hypoxia)and plateau hypoxia+TAK-071(an agonist of M1AChR)group.Morris water maze was utilized to detect the learning and memory ability of rats.Immunohistochemistry was utilized to determine the distribution and expression levels of M1 AChR,GluA1,and GluA2 in hippocampal CA1 and CA3 areas of rats.Western Blot was utilized to determine the expression levels of M1 AChR,GluA1,p-GluA1(Ser845),GluA2 and p-GluA2(Ser880)in hippocampus of rats.RT-qPCR was utilized to determine the mRNA expression levels of M1 AChR,GluA1,GluA2,ionotropic glutamate receptor NMDA type subunit 2A(NR2A),tau and amyloid β-protein(Aβ)in hippocampus of rats.Results:Plateau hypoxia could significantly increase the escape latency of rats in the spatial navigation test and shorten the stay time of rats in the target quadrant in the spatial probe test,while TAK-071 treatment could significantly improve the memory status of rats damaged by plateau hypoxia in the spatial probe test.There was a good correlation between the expression of GluA1,GluA2 and M1 AChR in CA1 and CA3 regions of hippocampus.In hippocampus,the expressions of GluA1 and GluA2 were synchro-nized with their corresponding phosphorylated proteins,and the correlativity between GluA1 and GluA2 and M1 AChR changed due to plateau hypoxia was reversed after TAK-071 treatment.TAK-071 could significantly reduce(P＜0.05)the expression levels of M1 AChR,GluA1,GluA2,NR2A,tau and Aβ mRNA increased due to plateau hypoxia.Conclusion:Activating M1 AChR can improve the memory impairment of rats caused by plateau hypoxia by regulating the expression of GluA1,GluA2 and their phosphorylated proteins.

Objective This study explored whether EPAS1 gene is involved in the proliferation of pulmonary arterial smooth muscle cells (PASMCs) during hypoxia when EPAS1 gene expression was interfered by small interfering RNA (siRNA).Methods The rat primary pulmonary artery smooth muscle cells were cultured and identified by immunofluorescence.The specific lipidosomes of EPAS1 siRNA were constructed and transfected into PASMCs,and the best targets were selected from the three interfering targets.The transfected PASMCs were cultured in hypoxia (37℃,5％ CO2,2％ O2) or normoxia (37℃,5％ CO2,20％ O2) for 24h,48h and 72h,respectively.The PASMCs proliferation was detected by CCK-8 assay.The protein expression of EPAS 1 and vascular endothelial growth factor (VEGF) were determined by Western blotting to investigate the effect of silencing EPAS1 gene expression on the proliferation of PASMCs under hypoxic condition.Results The specific liposomes ofEPAS1 siRNA were successfully constructed and transfected into PASMCs,and the best interfering target were selected from the three interference targets.The proliferation of PASMCs was increased and the protein expression of VEGF was up-regulated in the PASMCs under hypoxic condition.Under hypoxic or normoxic condition,PASMCs proliferation and VEGF protein expression of PASMCs were suppressed by EPAS 1 siRNA.Conclusion EPAS 1 gene might be involved in the proliferation of rat PASMCs by regulating VEGF protein level under hypoxic condition.

Objective To summarize the experience in diagnosis and treatment of primary splenic tumor. Methods The clinical data of 43 cases of primary splenic tumor patients treated in our hospital between January 1990 to January 2003 were retrospectively analyzed. Results The detectable rate of B-US was 95.3% and that of CT was 96.6%. All the 43 patients included 28 with benign tumors and 15 with malignant ones. The median follow-up period was 6.8 years(2 months-15 years). The 5-year overall survival of benign tumor was 100% and The 1-,3-and 5-year overall survival rates of malignant tumor were 80%, 53.3 % and 26.7%, respectively. Conclusion The image methods such as B-US and CT are the main diagnostic methods for primary splenic tumor. Early diagnosis, radical operation and combined therapy are important for improving the prognosis of the tumor.

Objective To investigate the change of aquaporin 2 (AQP2) mRNA and protein levels in renal collecting duct of SD rats after hypoxin caused by rising of the altitude to 4600 m. Methods Forty male SD rats were randomly divided into 4 groups (24 h, 48 h, 72 h and 1 week group), and 10 rats in Xining city were used as control group. All the 40 SD rats were transported to Kekexili Natural Reservation areas (4600 m) in Qinghai province. Rats of four experimental groups were sacrificed and renal tissue samples were harvested at different time point respectively, the control group rats were treated in Xining city (2260 m) as well. The concentration of plasma antidiuretic hormone (ADH) was measured by radioimmunity method. The expression of AQP2 mRNA and proteins was evaluated by real-time fluorescent quantitative-PCR, Western blot and immunofluorescence assay. Results The concentration of plasma ADH was decreased at 24 h and was only 28.5% of that of control group, reaching the lowest concentration at 48 h [(86.94±6.49) μg/L vs (302.5±310.48) μg/L], then it increased gradually and was similar to the control group at 7 d [(306.46±11.14) μg/L vs (302.53±10.48)μg/L, P＞ 0.05]. There were significant differences of the control group with 24 h, 48 h and 72 h groups, respectively[(302.53± 10.48) μg/L vs (142.46±10.57)μg/L, (86.94±6.49)μg/L, (169.65±11.15) μg/L respectively, P＜0.01]. The change of AQP2 gene expression level was consistent with the change of ADH. It was decreased at the begining when exposure to altitude and it reached its lowest level at 48 h. It was then returned to high level similarly to that of the control group at 7 d (0.09±0.01 vs 0.09± 0.008, P＞0.05 ). There were significant differences of the control group with 24 h, 48 h and 72 h group, respectively (0.09±0.008 vs 0.04±0.005, 0.03±0.002, 0.04±0.003 respectively, P＜0.01 ). Conclusions AQP2 expression in the renal collecting duct of SD rats is altered over the period exposed to altitude. It is decreased in the early hypoxia period, and is increased in later period. This change may be related to the intensity of hypoxia, which is mediated by a potential adaptation mechanisms against hypoxia caused by high altitude.