Objective　To establish the method of 2-dimensional electrophoresis(2-DE) for proteins of human spermatozoa and to construct a protein map of human spermatozoa. Methods　The sperm pellet was prepared with simple Percoll layer protocol. We studied the effects of various sample preparation methods, loading quantities and isoelectric-focusing protocols on the quality of silver-stained 2-DE map, and constructed a primary protein map of human spermatozoa. Result　Up to 703 protein spots were acquired with sample preparation Method Ⅰwhile only 194～210 spots with Method Ⅱ.With immobilized pH gradients and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(IPG-DALT) we could acquire over 700 spots while only 280～300 with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(ISO-DALT). Conclusion　It is satisfactory to lyse sperm with sample preparation Method Ⅰ and to separate sperm proteins by IPG-DALT for establishing 2-D map of human sperm.

Objective To analyze the chromosome abnormalities in the patients with primary and secondary amenorrhea in the infertility clinic, and provide the guidance for their reproduction.Methods Peripheral blood lyphocytes chromoseme samples were prepared and the karyotype was analyzed according to routine method.Results 24 abnormal chromosome karyotypes were detected in 108 cases, the abnormality detection rate was 22.2%, and it was mainly involved in the numerical or structural abnormality of chromosome X. Conclusions Chromosome abnormality is an important cause for primary and secondary amenorrhea, and the X chromosome abnormality is involed mainly.For those patients,chromosome examination should routinely be taken before assistant reproductive technique.

In line with guidelines of the health reform and realities in China,we comprehensively analyzed the strengths,weaknesses,opportunities,and threats in improving the hierarchical medical system by means of the SWOT method.Based on such,we proposed some corresponding development countermeasures,for policy and decision references in developing a medical system of reasonable division of responsibilities.

Objective:To explore the usage of system dynamics analyzing the influencing factors of total health expenditure(THE),forecast the trends of total health expenditure based on the method of System Dynamics and put forward relative countermeasures.Methods:According to the literature and expert consultation,the population,elderly,GDP,government health expenditures,drug costs and the number of health workers per thousand population were the important factors for THE.These factors were incorporated into system dynamics model for total health expenditure and conducted the simulation.Results:Based on the examination,the simulation results highly corresponded to historical data (2002-2014) and the official forecast (2015-2020),which showed that the model had good stability and reliability.The simulation results showed that steadily increasing total health expenditure would reach 7 457.12 billion yuan in 2025.Conclusion:The system dynamics approach had superior characteristics to other projection methods in terms of stability and reliability.In order to control the irrational growth of total health expenditure,it needed to increase the prevention investment,decrease the incidence of chronic diseases;sustainably improve the reform of public hospitals,implement the basic drug system;change the income-oriented performance appraisal to realize the labor value of medical staffs.

We present a case of two-year old boy with solitary atypical adenomatous hyperplasia (AAH) in extralobar sequestration (ELS), which was misdiagnosed as diaphragmatic hernia before surgery. Review of AAH and pulmonary sequestration (PS) revealed that the present case is the youngest of solitary AAH and also the first report of solitary AAH arising in ELS without a primary lung cancer. In a sense, the present case firstly supports the hypothesis that ELS may be an underlying cancer predisposition syndrome, so aggressive surgical therapy should be recommended for ELS.

ObjectiveTo summarize the experience of diagnosis and different surgical management for Nevin Stage V gallbladder cancer. Methods The present study was an retrospective analysis of 74 patients who were operated in our hospital and suffered from Nevin Stage V gallbladder cancer proved histopathologically. ResultsThe diagnostic accuracy of color Doppla ultrasound was 75.0%(21/28), abdominal enhanced CT, 82.1%(32/39).23 patients with Nevin Stage V disease received radical or extended radical cholecystectomy, the mean survival time was 1 year and 3 months;11 patients with Nevin Stage V disease without a complete resection had a mean survival of 6 months; 38 patients with Nevin Stage V diesase had a mean survival of 2 months after palliative bypass procedure or biopsy. ConclusionA exploratory operation by laparoscope is useful in the diagnosis of advanced gallbladder cancer. According to local situation of gallbladder cancer and general body state, different management are employed in treatment so that the effect may be improved.

Objective To analyze the effect of sperm factor on recurrent pregnancy lass.Methods Accordingto WHO criteria,we routinely analyzed the sperm of 21 unexplained recurrent pregnancy loss patients as study group.and 20 proven fertility men as control group.Meanwhile,we evaluated sperm chromosome aneuploidy for chromosomes 18,X and Y using standard FISH procedure,and sperm DNA ingegrity using sperm chromatin structure assay(SCSA)procedure.Results The percentage of normal morphology sperm is(10.8±2.8)%,and DFI value is(21.7±6.7)%,and the percentage of sperm aneuploidy for sex chromosome and chromosome 18 is(0.80±0.14)%and(0.23 ±0.09)%respectivelyin study group.The differences between the two groups were statistically significant(P＜0.01).Conclusion Sperm morphology,DNA integrity and chromosome aneuploidy are the correlative factors to recurrent pregnancy loss.

Objective To explore the prospects for applying a brain-computer interface system in the reha-bilitation of dyskinesia by testing the brain-electricity signals of spinal cord injury patients and stroke patients during motor imagery. Methods The brain-electricity signals of 5 healthy people, 4 spinal cord injury patients and 2 stroke patients were tested during motor imagery, and the results were analyzed with a computer. Results All of the 5 healthy people showed event-related synchronization and desynchronization (ERS/ERD) in the α and β bands in the motor related cortex. Three of the 4 spinal cord injury patents showed similar characteristics, and 1 showed ob-vious and stable characteristics, but the 2 stroke patients did not show meaningful characteristics. Conclusions Spinal cord injury patients have brain-electrlcity characteristics similar to those of healthy people when they are expe-riencing motor imagery, whilst in the affected hemisphere of stroke patients the brain-electricity characteristics are not similar to those of healthy people.

Objective To investigate the effects of Annexin-A1 ( Anxa1 ) gene silencing induced by siRNA on the growth and migration of microglial BV-2 cells and its possible mechanisms.Methods A synthesized siRNA duplex targeting Anxa1 gene was transfected into BV-2 cells.The efficiency of siRNA-in-duced Anxa1 gene silencing was evaluated on both mRNA and protein levels by using reverse-transcription PCR and Western blot assay.MTT assay was performed to measure the proliferation of BV-2 cells with si-lenced expression of Anxa1 gene.Flow cytometry with Annexin V-FITC/PI double staining was used to de-tect the apoptosis rate of BV-2 cells.Transwell chambers were used to analyze the effects of siRNA-induced Anxa1 gene silencing on the migration of BV-2 cells.Western blot assay was performed to detect the expres-sion of signaling proteins related to cell cycle and migration.Results Compared with the siRNA negative control ( siRNA-NC) group, the inhibitory rates of siRNA-induced Anxa1 gene silencing on the proliferation of BV-2 cells were significantly increased at the time points of 24 h, 48 h and 72 h after intervention [(16.9 ±2.1)%, (23.1±3.6)%and (42.4±1.7)%vs (1.35±0.5)%, (2.06±0.7)% and (8.65±0.9)%, P<0.05 ].The apoptosis rate of BV-2 cells transfected with Anxa1 siRNA was (18.4±2.1)%, which was significantly elevated as compared with that of the siRNA-NC group (5.2±0.3)%and control group (4.3±0.2)%.Cell migration of the Anxa1 siRNA transfected BV-2 cells was inhibited remarkably at 48 h as com-pared with that of the siRNA-NC group (28.7±5.2 vs 173.4±11.4, P<0.01).Moreover, the suppressed expression of Cyclin D1 protein and activation of p38 and JNK signaling pathways were induced by silenced expression of Anxa1 gene in BV-2 cells.Conclusion The growth and migration of BV-2 cells were signifi-cantly inhibited by silencing the expression of Anxa1 gene with siRNA, the possible mechanisms might be associated with the suppressed expression of Cyclin D1protein and the activation of p38 and JNK signaling pathways.

Objective To investite the effect of endotoxin pretreatment on lung injury induced by hepatic ischemia reperfusion in rabbits and its mechanism. Method Forty-eight New Zealand white rabbits were randomly divided into4 groups with 12 rabbits each group:routine control group,pretreatment control group,ischemia reperfusion group (IR group), and preperfusion group( LPS + IR group). Rabbits of routine control group received operative dissector only, and those of pretreatment control group received pretratment of daily intraabdominal injection of lipopo|ysaccharide(O.5,0.5,and 1.0 mg/kg,respectively)in the 3 days before operative dissector.Livers of IR group were rendered and ischeraic for 30 minutes, and repeffused for up to 4 hours. Rabbits of LPS +IR group received the preueaunent before heretic ischemia repeffusion. Four hours after reperfusion, serum endotoxin,tumor necrosis factor-α(TNF-α), wet/dry ratio and broncho-alveolar lavage fluid protein content of lung,malondialdehyde(MDA) and mpenrxide dismutase(SOD) in lung homogenate, lung injury ratio, and activity of Nuclear factor-kB(NF-kB) in alveolar macrophage wene examined. Differences within the groups were analyzed using One way ANOVA. Results Between the two control groups,there were no significant differences in all indexes(P>0.05). The TNF-α[ (48.31±5.31)pg/ml vs.(56.47±5.09)pg/ml, P<0.01],wet/dry ratio [(4.98±0.33)vs. (5.22±0.31), P = 0.03],broncho-alveolar hvage fluid protein content[(0.68±0.11)g/L vs. (0.76±0.10)g/L, P =0.04],MDA[(0.86±0.06)nmol/mg vs. (0.93±0.07)nmol/mg, P =0.02],lung injury ra-tio[(13.4±4.3)% vs. (17.4±4.1)%, P = 0.03],and the activity of NF-gB[(5.82±1.12)OD/mm2 vs.(7.40±1.26)OD/mm2, P<0.01] in alveolar macrophage of the LPS+ IB group were all significantly lower than those of IB group, while the SOD[ (90.30±7.38 )U/rag vs. (84.44±7.90 )U/rag, P = 0.04]of LPS + IR group was significantly higher than that of IR group. Conclusions Endotoxin pretrealment may ameliorate the lung injury induced by hepatic isehernia reperfusion. The mechanism may be that endotoxin pretreatment deoreases production of serum TNF-α and the activity of NF-kB in alveolar maerophage.