Mammalian target of rapamycin( mTOR) is a key reg-ulator of aging and aging-related diseases. Rapamycin ( RAPA) induces and promotes the process of cell autophagy through in-hibiting mTOR pathway. Autophagy exerts a crucial role in main-taining the cellular meostasis, which provides essential materials for cell reconstruction, regeneration and repair via degradating the redundant, damaged, or senescent proteins and organelles. Hutchinson Gilford progeria syndrome ( HGPS ) patients are al-ways accompanied with abnormally accumulated progerin in cells. Similar to HGPS, abnormal protein accumulation is the common pathological feature of neurodegenerative diseases, in-cluding Huntington′s disease, Parkinson′s disease, Alzheimer′s disease and so on. Degradation of these abnormal proteins pre-dominantly depends on cell autophagy. Thus, rapamycin is a po-tential anti-aging drug for HGPS and aging-related diseases thera-py. This view focuses on the effects of rapamycin on cell autoph-agy and clinical application in HGPS and neurodegenerative dis-eases.

Objective:To study the expression of TCRV? subfamily which specially recognize the hepatoma cell antigen and the apoptosis of hepatoma cell induced by McAb costimulated PBLs.Methods:The change of the phenotype of PBLs was studied by flow cell cytometry and the level of the expression of TCRV? was studied by RT-PCR and Southern blot,the PTK by western blot.The hypermicroscopic ultrastructure was observed through transmission electron microscope.Results:The level of CD3 and CD8 of PBLs was significantly increased after acted with hepatoma cells,while there was no change in CD4.The expression of TCRV?7 of PBLs was dramaticly increased and peaked at 4 days.PTK increased correspondently,to 58% compared with 11% in control.Besides anti-CD3 McAb induced lymphocyte apoptosis,the mediated apoptosis of hepatoma cells was found in the other three groups.Conclusion:TCRV?7 was the tumor antigen specific T cell receptor,and it activate the PTK signal pathway.The McAb activated lymphocytes initiated apoptosis in hepatoma cells.

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 study the biological effects of TCR to hepatoma cell by transfection V?7 to lymphocytes. Methods:TCRV?7 gene was amplified by RT PCR and cloned to expression vector pLXSN. The recombinant was transferred into lymphocytes by Lipfectin Reagent transfection,then the lymphocytes were co cultured with hepatoma cells.The phenotype of lymphocytes was detected on the Flow Cytometry and the ultrastructure of the hepatoma cells was showed by electronic microscope.Results:The lymphocyte amount with TCRV?7 expressing in those being transfected was much more than those no transfection.Apoptosis appeared in the hepatoma cells.Conclusion:TCRV?7 subfamily can recognize hepatoma antigen and stimulate T cell.

Objective: To explore the clinical, pathological and echocardiographic characteristics of rare primary non-mucinous cardiac tumor (PNCT) and to investigate the diagnostic value of echocardiography in benign and malignant PNCT with various pathological classiifcations. Methods: We retrospectively studied 32 PNCT patients including 21 benign and 11 malignant patients treated in our hospital from 2003-01 to 2013-02. There were 23 male and 9 female from 0.5 to 66 (36.92 ± 20.17) years of age. We analyzed the clinical information of echocardiography, CT, MRI and operation, and the ifndings were conifrmed by pathology. Results: Statistic analysis presented that there were more male PNCT patients than female, the ratio of male/female in benign patients was 16/5, in malignant patients was 7/4, P<0.05, the age was similar between 2 groups, (38.06 ± 21.68) years vs (35.72 ± 13.55) years,P>0.05, and either infant or adult could suffer from PNCT. Benign PNCT was more in left heart and malignant PNCT was more in right heart. Compared with benign PNCT, malignant lesions were usually no base, with irregular shape, pericardial effusion and surrounding tissue adhesion. Conclusion: Echocardiography was sensitive for diagnosing the patients with PNCT, it could preliminarily identify benign and malignant lesions with different ultrasonographic manifestation.

AIM To investigate the effects of Sanqi Huayu Oral Liquid (Notoginseng Radix et Rhizoma,Corydalis Rhizoma) on femoral shaft comminuted fracture caused by trauma in perioperative period of high-risk groups.METHODS One hundred and twenty patients meeting the inclusion criteria were randomly and equally divided into treatment group and control group.The control group was treated by low molecular weight heparin calcium injection,while the treatment group was additionally given Sanqi Huayu Oral Liquid.Two groups' levels of serum IL-6,TNF-α and D-D were detected on pre-operation and the 1 st,3rd,7th after operation,respectively.Additionally,the adverse drug reactions were also observed.RESULTS There were no statistically significant differences in the levels of serum IL-6,TNF-α and D-D between the two groups before operation.The first day after operation,the levels of serum IL-6,TNF-α and D-D between the two groups showed increasing trends,which had similar elevated levels without statistically significant differences.The third day after operation,the levels of serum IL-6,TNF-α and D-D between the two groups presented downward trends,but those in the treatment group were decreased more markedly with statistically significant differences.Furthermore,the seventh day after operation,the levels of serum IL-6,TNF-α and D-D between the two groups were lower than those before the operation,and these indices in the treatment group were lower than those in the control group with statistically significant differences.CONCLUSION Sanqi Huayu Oral Liquid can not only effectively reduce the levels of serum IL-6,TNF-α and D-D in perioperative period of high-risk groups,but also prevent the formation of deep vein thrombosis.

Objective:To investigate the effects of attention and interpretation therapy on fatigue and sleep quality in patients with gastrointestinal tumor during chemotherapy.Methods:From December 2018 to December 2019, eighty-four patients with gastrointestinal tumor hospitalized in two hospitals (third-grade class-A) in Ningxia were selected as the research objects. According to the random number table, they were divided into the control group and the observation group with 42 cases in each group. Patients in the two groups received routine care. On this basis, the observation group received 10 weeks of attention and interpretation therapy. Cancer patients fatigue scale and Pittsburgh sleep quality index(PSQI) scale were used to evaluate before intervention, 10 weeks after intervention and 3 months after intervention. SPSS 17.0 software was used for statistical analysis. Independent sample t-test and repeated measure analysis of variance were used for comparison between groups. Results:(1) The time effect, inter group effect and interaction effect of fatigue total scores and each dimension score of the two groups were significant (all P<0.01). Further simple effect analysis showed that there were significant differences in the total score and each dimension score of fatigue between the control group and the observation group at 10 weeks after intervention and 3 months after intervention (all P<0.01). (2) The time effect, inter group effect and interaction effect of PSQI total score, sleep quality, sleep time, sleep disorder score were significant (all P<0.01), but the time effect, inter group effect and interaction effect of sleep efficiency, daytime dysfunction and hypnotic drug use score were not significant (all P>0.05). Further simple effect analysis showed that there were significant differences in PSQI total score, sleep quality, sleep time, sleep time and sleep disorder scores between the control group and the observation group 10 weeks after the intervention(PSQI total score (6.83±2.46) vs (10.79±1.01); sleep quality (1.00±0.22) vs (1.24±0.82); sleep time (0.91±0.26) vs (1.40±0.86); sleep time (1.00±0.20) vs (2.02±0.72); sleep disorder (0.79±0.22) vs (1.60±0.59) and 3 months after the intervention(all P<0.01). Conclusion:Attention and interpretation therapy can effectively alleviate the fatigue of gastrointestinal tumor patients during chemotherapy, and improve sleep quality.

Objective:To investigate the effects of overexpression of long non-coding RNA maternally expressed gene 3 (LncRNA MEG3) on autophagy, apoptosis and mammalian rapamycin target protein (mTOR) pathway in pancreatic cancer cells (PANC1) .Methods:The pCMV-N-Flag-MEG3 expression plasmid was constructed and transfected into PANC1 cells. The expression of LncRNA MEG3 in hpde6c7 (normal pancreatic cells) group, PANC1 (blank control) , Vector (PANC1 cell transfected empty vector) group and MEG3 (PANC1 cell transfected with pCMV-N-Flag-MEG3 recombinant plasmid) group was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) ; methyl thiazolyl tetrazolium (MTT) , flow cytometry and monodansylcadaverin (MDC) staining were used to detect the effects of overexpression of LncRNA MEG3 on the proliferation, apoptosis and autophagy of PANC1 cells; Western blot was used to detect the effects of overexpression of LncRNA MEG3 on the expression levels of Bcl-2, Bax and Beclin-1 in PANC1 cells, and the phosphorylation levels of mTOR, ribosomal p70S6 kinase protein (SK61) and uclear initiation factor 4E binding protein 1 (4E-BP1) in mTOR pathway.Results:Compared with those in PANC1 group and Vector group, the expression level of LncRNA MEG3 (0.36±0.08 vs 0.35±0.11 vs 0.69±0.09) , proliferation inhibition rate (3.35%±0.12 vs 3.23%±0.09 vs 36.77%±0.13) , autophagy rate (29.32%±1.03 vs 26.73%±1.32 vs 57.76%±1.09) , apoptosis rate (9.85%±1.58 vs 9.73%±1.12 vs 35.89%±1.05) , expression levels of Bax (0.26±0.08 vs 0.29±0.05 vs 0.83±0.08) and Beclin 1 (0.15±0.06 vs 0.17±0.02 vs 0.61±0.03) of PANC1 cells in MEG3 group were significantly higher (all P<0.05) , and the expression level of Bcl-2 (0.79±0.12 vs 0.81±0.09 vs 0.30±0.03) and phosphorylation levels of mTOR (1.08±0.05 and 1.06±0.08 vs 0.37±0.10) , SK61 (1.12±0.06 and 1.11±0.09 vs 0.41±0.03) and 4E-BP1 (0.97±0.07 and 0.95±0.03 vs 0.39±0.05) in mTOR pathway were significantly lower (all P<0.05) . Conclusion:Overexpression of LncRNA MEG3 can inhibit the proliferation of PANC1 cells, promote apoptosis and formation of autophagic vesicles, which may be related to the blocking of mTOR pathway.

Objective To investigate the role of Notch signaling pathway to maintain the stem cell-like characteristics of osteosarcoma and its underlying mechanism.Methods Lentiviral NICD1 or Numb-shRNA was transduced into MG63 osteosarcoma cells to activate Notch activity in vitro.The impact of Notch on osteosarcoma stem cells were assessed by the tumor sphere formation assay and flow cytometry analysis of cell surface markers STRO-1/CD117.The expression of stem cell related genes (Sox2,Oct4) were evaluated by Western blot and qPCR.The nude mice were randomly divided into 3 groups:the NICD1 overexpression (NICD-OE) group,the DAPT group and the control (CON) group.The tumor growth was monitored for 8 weeks and the tumor volume and weight were recorded weekly.To investigate whether Notch regulates Eph pathway,Eph pathway related protein EphB,pEphB was measured by Western blot.The impact of ephrinB 1 on NICD overexpression cell were assessed by tumor sphere formation assay.The expression of Sox2 and Oct4 was evaluated by Western blot.Results NICD1 overexpression or Numb-shRNA increased the activity of Notch pathway.The Notch-activated osteosarcoma showed enhanced in vitro tumor spheroid formation capacity,increased Stro-1/CD117double positive ratio,and upregulated expression of Sox2 and Oct4 in vitro.In animal experiments,it was found that activation of Notch pathway promoted tumor formation in vivo and Notch inhibition decreased it.The primary osteosarcoma cells were obtained from mice xenograft treated with DAPT and its tumor sphere formation capacity was significantly reduced.Finally,The Notch pathway inhibits the phosphorylation of EphB,as well as the downstream signal pathway of EphB,but there is no significant change in total EphB.The activation of Eph pathway inhibited Notch induced up-regulation of tumor sphere formation and Sox2 and Oct4 expression.Conclusion Notch signaling pathway maintains the stem cell-like characteristics of osteosarcoma probably by inhibiting the Eph pathway.