Objective To investigate the remote liver injury induced by reactive oxygen species (ROS) after intestinal ischemia/reperfusion (I/R) and the mechanism of liver protection afforded by methylene blue (MB) Methods Thirty-two rabbits weighing 2.3-2.9 kg were randomly divided into four groups: (1) normal group in which sham operation was performed; (2) I/R group in which intestinal I/R was produced by clamping anterior mesenteric artery for 1 hour; (3) in treatment group A MB 3 mg?kg-1 was given iv immediately ofter before declamping and (4) in treatment group B MB 15 mg?kg-1 iv was given. Common carotid artery was cannulated for direct continuous blood pressure monitoring. Blood samples were taken before anterior mesenteric artery occlusion, immediately after and 30 min, 1 h and 2 h after decalmping for determination of blood MDA concentration. At the end of the experiment a small piece of hepatic tissue was obtained for determination of its superoxide dismutase (SOD), catalase (CAT), xanthine oxidase (XO) and MDA levels.Results In I/R group blood MDA level increased significantly after I/R as compared with the baseline value before I/R and blood pressure decreased significantly after I/R, while in treatment group A and B there was no significant increase in blood MDA level after I/R. In I/R group liver MDA level was significantly higher than that in normal group, but there was no significant difference in liver MDA level between normal group (group 1) and treatment groups (group 3,4) .Conclusion Methylene blue can reduce the generation of ROS during intestinal I/R and protect liver from ROS-induced injury.

Objective To investigate the pattern of outpatients traffic,orchestra service resources,and optimize service provision,for a harmonized outpatient service.Methods A baseline survey was made at the outpatient department in 30 days(six hours per day,7:00-11:00,and 13:00-15:00).On such basis,data of the hospital's HIS system within the last five years were called into play for statistics analysis and simulation test,for the purpose of shift scheduling and resources allocation optimization.Results This study discovered each January-February as the valley of clinic visits and each July-August as the peak.52-week data analysis of a year found that the days of daily registrations between 4000-5000 are up to 163 days,those over 5000 are up to 86 days,of which Mondays with over 5000 registrations are up to 35 days.Each Monday is found to be a peak of outpatient visits,with significant drop during Saturdays and Sundays; daily registrations peak between 7:00-10:00 in the morning,while the traffic rises again during 13:00-14:00 in the afternoon,yet up to 26 ％of the morning peak only.Conclusion Statistics analysis of the outpatient service and emergency service at the outpatient department can help identify their patterns of operations,for effective improvement of service provision,alleviation of peak hour workload,shorten patient waiting time and better patient satisfaction.

This study is to investigate the amelioration effect of glucocorticoid receptor (GR) antagonist mifepristone on the changes of learning and memory abilities in rat model of depression. In the present study, a 35-day rat chronic unpredictable stress (CUS) model was used to observe both depression-like behaviors with sucrose preference test and open-field test and learning and memory-associated behaviors with Morris water maze test. A total of 45 male adult Sprague-Dawley rats were randomly assigned to three groups of equal size: control group (CON); CUS group (CUS); CUS + mifepristone group (CM). Animals in CM group were first exposed to CUS for 14 days, and then were administered with 50 mg x kg(-1) x d(-1) of mifepristone with continued CUS procedure. Corticosterone EIA Kit was used to detect the concentration of plasma corticosterone (CORT). Nissl staining was used to observe the structure of hippocampus. The results demonstrated that CUS exposure induced both depressive-like and learning and memory-associated behaviors and these deficits were reversed by mifepristone. Compared to CON group, the concentration of plasma CORT increased significantly in CUS group. CUS exposure damaged the structure of hippocampus, whereas mifepristone had an amelioration effect. Together, the structural deficits of hippocampus resulting from long-term stress exposure, which could contribute to the impairment of learning and memory in depression, are reversed by the GR receptor antagonist mifepristone.

Abstract:Aim To study the effect of rotenone on α-synuclein in rat midbrain of Parkinson's disease(PD).Methods Rats were subcutaneouly injected with chronic low dose rotenone(1.0 mg·kg~(-1)·d~(-1)).Movements within 5 minutes were evaluated in open field test.Tyrosine hydroxylase(TH)-positive neuronsin the midbrain were measured with immunofluorescence staining.α-SYN protein level in the midbrain was demonstrated with immunohistochemical staining and Western blot.Results Compared to the control group,latency time,crossing,rearing and rearing time were changed significantly(P<0.05,0.01)in the rotenone group,TH-positive neurons were reduced(P<0.05)and α-SYN protein level in the midbrain was increased(P<0.05).Conclusion Injection with rotenone can induce PD symptom,which may be correlated to α-SYN protein level in the midbrain.

Aim To construct the cell model targeted on the damage by α-synuclein for screening anti-Parkinson’s Disease (PD)compounds.Methods The cDNA fragment of α-synucle-in gene was obtained by PCR methods and inserted into the re-combinant prokaryotic plasmid by molecular cloning technique. The recombinant plasmid was transformed into Escherichia coli, and subsequently induced to express α-synuclein protein.The recombinant α-synuclein was purified and identified by affinity chromatography,immunoblotting and mass spectrometry.The cells damage by α-synuclein was evaluated through cell viability measured by 3-(4,5-dimethyl-2-thiazolyl )-2,5-diphenyl-2-H-tetrazolium bromide.Results The obtained cDNA fragment ofα-synuclein in accordance with its theoretic molecular weight was cloned into pET30a plasmid and verified by sequencing.The re-combinant plasmid was transformed into bacteria E.Coli.BL21 (DE3)and induced to express α-synuclein by isopropyl β-D-1 -thiogalactopyranoside (IPTG).The expression condition was op-timized according to the culture temperature,the concentration of IPTG and the proliferation state of bacteria.The purified α-synu-clein was proved to be a 1 5.3 ku molecule weight protein,and could be immunoblotted with anti-α-synuclein antibody.The pu-rified α-synuclein could decrease the viability of PC1 2 cells and primary neurons significantly,and its effect was in a concentra-tion-dependent manner.Conclusion We have succeeded in constructing the cell model targeted on the damage by α-synucle-in.

Aim To investigate the role of chemokine-like factor 1 ( CKLF1 ) in SH-SY5 Y cell migration and its molecular regulatory mechanism. Methods SH-SY5Y cells were stimulated with CKLF1 for 0. 5 h, 2 h, 8 h and 24 h, respectively. The migration distance and the percentage of migration cells were recorded by CELLocate analysis. The phosphorylation of focal ad-hesion kinase ( FAK) at Tyr-397 site was detected by Western blot analysis. By chemotaxis assays, we con-firmed the chemotaxis of CKLF1. Furthermore, FAK inhibitor PF-573228 and PLCγ inhibitor U73122 were used for the research of molecular regulatory mecha-nisms involved. Results CKLF1 promoted cell migra-tion and induced a strong increase in the phosphoryla-tion level of FAK-pY397 , which were significantly at-tenuated by the presence of U73122 ( a specific inhibi-tor for PLCγ) . In addition, the chemotaxis of CKLF1 was obviously blocked by the FAK inhibitor PF-573228 . Conclusion CKLF1 induces SH-SY5 Y cell migration via PLCγ/FAK signaling pathway.

Aim To study the effect of synapsinⅠon synaptic transmission in rat dentate gyrus induced by(-) clausenamide.Methods The basal synaptic transmission experiment was conducted through electrophysiological recordings.The effect of(-) clausenamide on synapsinⅠ phosphorylation was measured by western blot and confocal microscopy.Results(-)Clausenamide increased the population spike(PS) of hippocampal dentate gyrus.The phosphorylation of synapsinⅠ was increased both in cortex and hippocampus,the maximum effect was observed at 5 min in hippocampus and at 15 min in cortex.Furthermore,(-)clausenamide promoted the phosphorylation of synapsinⅠat a dose-denpendent manner in PC12 cells.The phosphorylation of synapsinⅠ in PC12 cells and synaptosomes incubated with(-)clausenamide was increased and reached maximum at 1~2 min.However,H89,PKA inhibitor,blocked the effect of(-)clausenamide on synapsinⅠ phosphorylation.Conclusion(-)Clausenamide activated synapsinⅠ via PKA signal pathway,which may contribute to the effect of(-)clausenamide on potentiating basal synaptic transmission.

Objective:To evaluate the effect of preoperative panel reactive antibody(PRA)levels on long-term survival after kidney transplantation. Methods:Data on 1 162 patients underwent first kidney transplantation performed between January 2001 and June 2014 were included in our center. According to the preoperative PRA levels,the patients were divided into negative group( PRA≤10%) and positive group( PRA>10%) ,which were retrospectively analyzed. Results: The 1-,5-,10-year patient survival rates of the negative group calculated by Kaplan-Meier were 96. 8%,89. 4%,78. 6%,respectively,while the positive group were 93. 5%,81. 6%, 65. 4%. The 1-,5-,10 -year death-censored graft survival rates of the negative group were 95. 9%,84. 8%,63. 1%,respectively,while the positive group were 92. 3%,74. 1%,51. 9%. The log-rank test revealed that there was significant difference between the patient and graft survival curves (χ2 =9. 623/11. 019, P=0. 002/0. 001 ) . Cox multivariate analysis found that preoperative PRA levels were independent risk factors for reducing the patient or graft survival rates(P<0. 001). Logistic multivariate regression analysis confirmed the significant association between preoperative PRA levels and the risk of acute rejection ( OR=8. 25,95% CI=2. 86-5. 72, P<0. 001). The 5-,10-year creatinine values were significantly lower in the negative group compared to the positive group(all P<0. 05), while there was no difference in the 1-year. In addition, Logistic multivariate regression analysis confirmed the significant association between preoperative PRA levels and the production of donor specific antibody(DSA)(OR=6. 89,95% CI=4. 52-9. 17,P<0. 05). Conclusion: The detection of preoperative PRA is an important indictor predicting the sensitivity status of the recipients. The preoperative PRA positive recipients need careful monitoring and diagnosis of acute rejection and DSA after kidney transplantation.

OBJECTIVE: To investigate clinical application of head impulse test with video recording eye movements in the diagnosis of vertigo. METHOD: The video head impulse test(vHIT) was used to measure the eye saccades and velocity gain in 95 patients with vertigo which were divided into two groups, peripheral vertigo (47 cases) and central vertigo(48 cases); the characteristics of eye saccades and velocity gain of six semicircular canals in different patients with vertigo were analyzed, and were compared between the two groups. RESULT: The vHIT result in patients with peripheral vertigo: in 22 patients (23 affected ears) with Meniere's disease, 21 ears were abnormal (91. 3%); the vHIT results in 4 patients with vestibular schwannoma, 2 patients with vestibular neuritis, 5 patients with delayed endolymphatic hydrops, 6 patients with sudden hearing loss accompanied vertigo, and 8 patients with vestibular dysfunction, were abnormal with correct saccades and/or lower velocity gain of vHIT. The abnormal vHIT results were also found in 35 of 48 patients (72. 9%) with central vertigo, which including posterior cerebral circulation ischemia(7 patients), cerebral infarction/stroke(6 patients), and dizziness with vertigo(17 patients) and others(18 patients). Abnormal rate of vHIT in patients with peripheral vertigo was 95. 7% (45/47), which was significantly higher than that (72. 9%) in patients with central vertigo. CONCLUSION It is easy to perform the vHIT which without adverse reactions. We can record high-frequency characteristics of vestibular-ocular reflex among six semicircular canals through vHIT. The vHIT results which show the function of vestibular ocular reflex in different diseases with vertigo, can help discriminate peripheral vertigo from central vertigo, and it is a practical assessment method for vertigo.
Dizziness ; Endolymphatic Hydrops ; diagnosis ; Head Impulse Test ; Humans ; Meniere Disease ; diagnosis ; Neuroma, Acoustic ; diagnosis ; Reflex, Vestibulo-Ocular ; Saccades ; Semicircular Canals ; Vertigo ; diagnosis ; Vestibular Neuronitis ; diagnosis ; Video Recording

Aim To study of the expression and distribution of four α-synuclein truncations in three cells.Method Four α-synuclein gene truncations were obtained by PCR method,followed by subcloning into the pEGFP-N1 eukaryotic expression vector.Four obtained recombination plasmids were transfected into MN9D cells,PC12 cells and SH-SY5Y cells using Lipofectamine 2000 respectively.The expression and distribution of four α-synuclein truncations were observed by Confocal.Results Distribution of four α-synuclein truncations was discrepant obviously,the truncations,with more C terminal remained,were prone to emerging in nuclei.Conclusion Localization of α-synuclein protein in cells may be related to the C terminal,and the whole C terminal plays an important role in distribution of α-synuclein into nuclei.