Objective To isolate and identify leukemia stem cells from acute myeloid leukemia patients for further research. Methods By Ficoll density gradient centrifugation, mononuclear cells were firstly separated from bone marrow of patients. According to specific surface markers, CD+34 CD+123 of leukemic stem cells were sorted by flow cytometer. Their ability of self-renewal and differentiation were evaluated by colony formation and cobblestone forming ability. At the same time, the purity and cell morphology of CD+34 CD+123 cells was analysed. Results Comparared with total mononuclear cells, the proportion of the CD+34 CD+123 cells after sorting was 10.7 %, and these cells showed the ability of colony forming and cobblestone forming, and the purity proportion of CD+34 CD+123 cells was 62.1%. Conclusion The leukemia stem cells were isolated successfully and could be used in further study.

Objective To analyze the negative experience from medical practice of medical students by Balint group case study,and to explore new approach for training doctor-patient communication skills.Methods 10 times of Balint group activities were held in Zhongshan Hospital of Fudan University,with one case each time.Grounded theory approach were applied for analyzing discussion record.Results The cases were occurred in doctor-patient interaction,teamwork,conflict between one's personal life and vocational development planning.Trigger events involved doctor-patient conflict,conflict of values,decision making problem while conffronted with incurable patients,role ambiguity,unfair treatment.Topics of discussion included emotional experience with patients,career development orientation,medical teamwork,clinical education of medical student.Conclusion Through practice of Balint group,medical students in teaching hospital can improve their awareness of complicating psycho-social problem of patients and themselves,as well as handling it.And it can improve the professional identity,professional growth and professional self-identity.

Objective To evaluate the effectiveness of using low-frequency repetitive transcranial magnetic stimulation (rTMS) along with donepezil in the rehabilitation of aphasia after cerebral infarction. Methods Fortythree aphasic patients with acute cerebral infarction were assigned into a treatment group or a control group.Both groups received standard basic treatment for acute cerebral infarction.From day 7 on,both groups received oral donepezil hydrochloride (5 mg/d) for 12 consecutive weeks.The treatment group received in addition 2 sessions of low frequency rTMS on the language center of the right hemisphere.The first session started at day 7 after onset,and each session consisted of rTMS on 10 consecutive days (once per day).There was a resting interval of 3 d between the two sessions.The rehabilitation of the aphasia was assessed prior to and after each session,and at the end of the entire 12 weeks of treatment using the western aphasia battery ( WAB ) scoring system and the aphasia quotient (AQ). Results There was no significant difference between the two groups after the first session in terms of the WAB or AQ results.However,the difference in improvement became statistically significant after the second session,as well as at the end of the entire 12 weeks.The average change in AQ was significantly greater in the treatment group. Conclusion Donepezil combined with rTMS early in the acute phase of cerebral infarction is helpful for the rehabilitation of aphasia.

Objective To investigate the therapeutic effects of repeated transcranial magnetic stimulation (rTMS) at high and low frequency for the rigidity and tremors of Parkinson's disease (PD).Methods Sixty-nine patients with PD received rTMS at 5 Hz or 0.5 Hz.The efficacy was assessed using the unified Parkinson's disease rating scale (UPDRS),motor function tests and motor evoked potentials (MEPs).Results Thirty-two patients with rigidity from PD received 5 Hz rTMS therapy,and their UPDRS total scores decreased significantly.Their motor function test results improved significantly and their MEP relaxed motor thresholds were significantly elevated.Thirtyseven patients with tremors from PD received 0.5 Hz rTMS therapy,and their UPDRS total scores were significantly decreased.Their motor function test results improved significantly and their MEP relaxed motor thresholds were significantly elevated.Conclusion High frequency rTMS may improve the symptoms of patients with PD rigidity and low frequency rTMS may improve the symptoms of patients with PD tremors.

Objective:To explore personality change and its association with lncRNA expression of peripheral blood mononuclear cells in patients with epilepsy.Methods:Fifty-eight epilepsy patients recruited by the convenient sampling were assessed utilizing personality diagnostic questionnaire(PDQ) for personality change screening.The expression levels of lncRNA in peripheral blood of study group and the controls were investigated by qRT-PCR.Descriptive statistical analysis, correlation analysis, regression analysis and ROC analysis were employed for data processing.Results:There were 9 of schizoid (S≥4), 11 of schizotypal(S≥5), 17 of paranoid (S≥4) and 15 of compulsive (S≥4) personality change in epilepsy patients, and 52 patients had different types personality changes(89.66%).Schizoid, schizotypal, paranoid and compulsive personality changes were negatively correlated with expression levels of NONHSAG012869(PR3), NONHSAT006265(PR4), ENST00000581634(PR6) and ENST00000524610(PR8) ( r=-0.46--0.71, P<0.05 or 0.01).PR1, PR3, and PR8 had significant predictive effects on schizoid personality change ( P<0.01), PR4, PR8 had a significant predictive effect on schizotypal personality change ( P<0.01), PR3, PR4 and PR6 had significant predictive effects on paranoid personality change( P<0.05), and PR4, PR5, PR8 had significant predictive effects on compulsive personality change ( P<0.05).The effects of lncRNAs on the personality change variance accounted for 0.36, 0.30, 0.40, 0.20 respectively.ROC curve analysis of the diagnostic value of lncRNA expression level on personality change in the epilepsy group showed that NONHSAG012869 (PR3), NONHSAT006265(PR4), ENST00000581634(PR6) and ENST00000524610(PR8) had certain diagnostic value for personality change.The area under curve(AUC)=0.650-0.682, P<0.05, 95% CI: 0.546-0.784. Conclusion:Schizoid, schizotypal, paranoid, and compulsive personality change are common in epileptic patients, and the expression level of peripheral blood lncRNA has a certain diagnostic value for personality change.

ObjectiveTo predict the target genes of miRNA associated with hsa_circRNA_102802 and hsa_circRNA_104597 using bioinformatics methods,and analyse the biological process and signaling pathway.MethodsTargetscan,miranda and mirbase three online database were used to predict target genes of miRNAs which are complementary to circRNAs.Target genes of miRNAs prediction results were taken their intersection of three online database,then take their collection as total target genes of miRNAs.The results were analyzed by gene ontology(GO) and KEGG pathway analysis using FunNet.Results199 target genes of has_miR_204_5p,has_miR_809,has_miR_520_5p,has_miR_423_5p,has_miR_617 and has_miR_877_5p were gotten from the intersection of 3 databases;and 410 targets of has_miR_659_3p,has_miR_9_5p,has_miR_661,has_miR_576_3p,has_miR_548d_5p,has_miR_548a_5p,has_miR_548b_5p,has_miR_876_5p and has_miR_744_5p were gotten from the intersection of 3 databases.GO analysis showed that Target genes involved in diverse biological processes including central nervous system,such as cortex development,axon guidance and extension,synaptic transmission,as well as the process of learning and memory (P<0.05).Enriched pathways were also revealed target genes involved in the signal path including the axon guidance,glutamic acid synapses,Wnt signaling pathway,ErbB signaling pathway,mTOR signaling pathway and VEGF signaling pathway,which connect closeiy with depression.hsa_circRNA_102802 and hsa_circRNA_104597 may play an important role in depression.Conclusionshsa_circRNA_102802 and hsa_circRNA_104597 may be associated with the pathologenesis of depression disorder.

AIM: To generate the angiopoietin-1 recombinant adenovirus for the further studies about adenoviral mediated angiopoietin-1 gene transfer in ischemic myocardium and its effect on the development of neoangiogenesis.METHODS: Angiopoietin-1-PAxCAwt cosmid DNA and adenoviral DNA-TPC were cotransfected to 293 cells by the use of calcium phosphate precipitation method. All recombinant adenoviruses were propagated and titrated in 293 cells, purified by cesium chloride density purification, dialyzed, and stored at-70 ℃. RESULTS: The result shows that the direction of the insert in the cosmid is correct and the replication-deficient angiopoietin-1 recombinant adenovirus was generated efficiently by COS/TPC homologous recombination, with the titers of 5.6?10 11 pfu/L. The viral stocks were demonstrated to be free of replication-competent wild type adenoviruses. The virus stocks in high titer were harvested in our experiment. CONCLUSION: The COS/TPC is an efficient method to prepare recombinant adenovirus and the angiopoietin-1 recombinant adenovirus could be further used in gene therapy.

Objective To study the prognosis in the patients with distant metastasis after nasopharyngeal carcinoma radical radiotherapy and to analyze its influencing factors .Methods One hundred and fifty patients with distant metastasis after nasopha-ryngeal carcinoma radical radiotherapy were followed up .Their clinical data were collected to conduct the prognosis analysis and in-fluencing factors analysis .Results The median survival time in the follow up cases was 13 .50 months ,the one-year survival rate was 49 .8% ,two-year survival rate was 31 .0% and three-year survival rate was 19 .3% .The Cox model single factors analysis re-sults showed that the N stage in initial diagnosis ,whether conducting chemotherapy in initial diagnosis ,time interval from radiother-apy finish to distant metastasis occurrence ,metastasis site ,chemotherapy after distant metastasis ,chemotherapy cycles and palliative radiotherapy were the prognosis related influencing factors (P<0 .05) ,in which the N stage ,metastatic site ,chemotherapy after dis-tant metastasis ,time interval from radiotherapy finish to distant metastasis occurrence and palliative radiotherapy were the inde-pendent influencing factors of prognosis(P<0 .05) .Conclusion The independent factors of prognosis in the patients with distant metastasis after nasopharyngeal carcinoma radical radiotherapy are the N stage at initial diagnosis ,metastatic site ,whether having chemotherapy after distant metastasis ,time interval from radiotherapy finish to distant metastasis occurrence and whether conduc-ting the palliative radiotherapy .

Objective To investigate the differential expression of microRNA (miRNA) in schizophrenia (SZ) patients,and explore the comorbidity of SZ and depression disorder based upon miRNA expression.Methods Affymetrix array analysis was used to investigate the differentially expressed miRNA in SZ patients firstly,and then quantitative real-time polymerase chain reaction (qRT-PCR) was further carried out to confirm the selected miRNA in peripheral blood mononuclear cells of 40 SZ patients,whom were administered by Positive and Negative Syndrome Scale (PANSS) and the selected miRNAs in depression disorder patients has also been confirmed by Affymetrix array analysis and qRT-PCR in our previous studies.Results Affymetrix array analysis indicated that there existed 33 miRNAs which differentially expressed (32 up-regulated and 1 down-regulated) compared with normal controls.qRT-PCR results suggested that the expression of 8 miRNAs (miR-1273d,miR-1303,miR-3064-5p,miR3131,miR-3687,miR-4428,miR-4725-3p and miR-5096) were significantly up-regulated in SZ;the miRNA differentially expressed in depression disorder patients also had differential expression in SZ patients (P＜0.05).There were significant correlation between the miRNAs differentially expressed in depression disorder patients and in SZ patients (P＜0.01).MiR-1972 differentially expressed in depression disorder patients had significant positive correlation with the positive symptoms of PANSS (P＜0.05),and miR-26b was positively correlated with composite factor (P＜0.05).Conclusion Comorbidity of SZ and depression disorder is observed not only on the clinical symptoms,but on the molecular genetic basis.

Objective To evaluate the maneuvre of curettage and aspiration(LTCA) in laparoscopic hepatectomy. MethodsWe used Peng′s multifunctional operative dissector(PMOD) to perform laparoscopic liver transection by maneuvre of curettage and aspiration in 20 cases undergoing laparoscopic hepatectomy. Results Procedures were all successful. The recovery was uneventful without any complications. Mean operative time was 105 minutes, the average bleeding volume was 420 ml, the largest excised sample size was 10 cm?9 cm?7 cm. All patients were discharged within one week. ConclusionsThe new technique-LTCA can be used in laparoscopic hepatectomy, it has the advantages of clear anatomy, good exposure of canal construction, rapid liver transection, satisfactory hemostasis and clear operative field.