The curriculum system of Proteomics was analyzed based on the teaching practice,the characteristics of ability training and gradation teaching were summarized and the prospect of curriculum optimization was proposed.These measures were conceived to enrich the course content and teaching methods for Proteomics course.

Objective To develop new methods to cultivate, retrieve and purify mouse mesenchymal stem cells (mMSCs). Methods Bone marrow was collected from 2-month-old Kunming mice by flushing femurs and tibias with complete medium of DMEM-LG. Cells were plated in a Petri dish. After 24 hours, non-adherent cells were removed by two to three washes with PBS, adherent cells were further cultured in complete medium and retrieved by trypsinisation with 0.25% trypsin for 5 min at 37 ℃. The treated adherent cells were cultivated with 3?dilution for further generations. CD11b-negative cells were retrieved from the collected adherent cells of 3rd generation by using immunomagnetic microbeads, and continued to be cultured in complete medium. After the cultured cells were retrieved, their morphology and their ability of osteoblastic differentiation and adipocytic differentiation were examined. Results Most of mMSCs from 1st generation were of shuttle shape, some of irregular shape. After treatment with magnetic microbeads and several generations, mMSCs were of spindle, star and irregular shape. These cells were of rich cytoplasma, clear nucleolus, and grew in parallel or vortex. The cultured adherent cells from the first and subsequent generations had plenty of CD11b-positive blooding-making cells. After 20-day osteoblastic induction, mMSCs differentiated into bone cells, which showed orange phosphate in extracellular matrix by Alizarin red S staining. mMSCs could differentiate into lipocytes. The size of cells increased along with fat-developing induction period. These cells showed many orange fatty follicles with O Red Oil dyeing. Conclusion Pure mMSCs can not be retrieved by either adhering method or generation cultivation method separately. The combined methods of adhering, immunomagnetic microbeads, and serial subcultivation is effective in vitro in retrieve mMSCs.

Objective To explore how to improve the immunogenicity of short epitope peptides of triggering melanoma MART 1 specific CD8 +T cell responses Methods Therapeutic peptides based on the immunodominant MART 127 35, HIV Tat49 57CCP sequence and a tetanus toxoid universal Th epitope were designed and synthesized The immunological functions were studied in PBMCs from HLA A2 + melanoma patients Results The results demonstrated that the peptides could trigger vigorous MART 1 specific CD8 + CTL activities in vitro The function of peptide containing MART 127 35 and tetanus universal Th epitope was more vigorous than that of MART 127 35 peptide, and the immunogenicty of the peptides with HIV Tat49 57CCP sequence, MART 127 35 and tetanus universal Th epitope was the most vigorous Conclusion Linkage of HIV Tat49 57CCP sequence and a tetanus universal Th epitope could dramatically improve the immunogenictiy of the MART 127 35 epitope peptide

Objective To explore how to trigger an HLA Ⅰ restricted CD8 + T cell response to exogenously synthesized peptides in vitro . Methods A new panel of therapeutic peptides based on the immunodominant B and CTL epitopes of HBV PreS 2 region and HBcAg and the tetanus toxoid common T helper epitopes were synthesized by Merrifield solid phase peptide synthesis, and HLA A2 + human PBMCs were used to investigate the immunological properties of the mimetic peptides. Results The results demonstrated that the peptides could trigger vigorous CD8 + HBV specific CTL responses in vitro specifically and effectively. Conclusion The results reveal that T helper plus B epitopes designing with the introduction of short and flexible linker can remarkably improve the immunogenicity of short peptides and hence produce effective CTL responses in vitro .

AIM: The aim of this work is to investigate the protective effects of Ginsenoside Rb 1(Rb 1) on apoptosis induced by hypoxia /hypoglycemia and reoxygenation in cultured rat hippocampal neurons. METHODS: Apoptosis were measured by flow cytometry; Morphological changes and neuronal necrosis were examined under microscope; The leakage of lactic dehydrogenase(LDH) and the product of nitric oxide(NO) were measured. RESULTS: hypoxia /hypoglycemia cultures for 5 h and reoxygenation induced neuronal apoptosis and necrosis,and significantly increased the leakage of LDH and the product of NO. The effects were enhanced with the extending time of reoxygenation. Rb 1 could significantly decrease the percentage of neuronal apoptosis and necrosis, and reduce the leakage of LDH and the product of NO. CONCLUSION: Rb 1 had an effect of anti-neuronal apoptosis. This effect might be related to the inhibition of the activity of NO synthase and NO production.

AIM: To analyze and identify the phosphoproteins associated with diazoxide preconditioning. METHODS: Proteomics technique was used to investigate the changes of phosphoprotein after diazoxide preconditioning. Adult rat ventricular myocytes were pretreated in the presence and absence of 200 ?mol/L diazoxide for 10 min. Phosphoproteins prepared and enriched respectively from control and diazoxide pretreated groups were then separated by two-dimensional (2D) gel electrophoresis and stained with sliver staining kit. Phosphoproteins of interest were further identified by mass spectrometry. RESULTS: Associated with diazoxide preconditioning, the proteins of chaperonin containing TCP-1 and hypothetical protein XP_346548 were phosphorylated significantly. The proteins of 94 kD glucose-regulated protein, calpactin I heavy chain and ferritin were dephosphorylated markedly (P

Objective To investigate the effects of extended nursing based on relationship, information and management on the self-care behaviors of patients with heart valve replacement. Methods According to admission time, 37 patients with heart valve replacement were set as the control group, which was given routine nursing intervention and another 42 patients as the study group, which received extended nursing based on the trinity of relationship, information and management 2~8 weeks after discharge. Before and after intervention, patients were evaluated by using the improved version of self-care behavior evaluation scale of heart disease (SNBAOHD). Result There were significant differences between the two groups in other items of SNBAOHD, except two items of low salt diet and medication (P<0.05). Conclusions Extended nursing after discharge based on the trinity of relationship, information and management can help the patients to be continuously cared. In this case, it is effective for the enhancement of off-hospital self-care ability and prevention of disease relapses.

Objective To evaluate the clinical value of apheresis platelets throught heanalysisof case control on the clinical efficacy and safety of cryopreserved apheresis platelets and fresh apheresis platelets.Methods 2 035 clinical cases of platelet transfusion in August 2014 to December 2016 by Using the closed loop intelligent path management and evaluation information system,456 cases were selected as control cases.Platelets were divided into the cryopreserved apheresis platelets group (group A,n=199) and fresh apheresis platelets group (group B,n=257) according to the transfused platelet type.The clinical application value of cryopreserved single platelets was evaluated by comparing the basic data,the effective indexes and safety indexes of the two groups.Results 1) The cases were 43.6％ (199/456) in A groups,and 56.4％ (257/456) in B groups,there were no significant difference in gender,age and medical and surgical cases between A and B group (P＞0.05);2) 199 cases in group A were cryopreserved platelets of 2 275 U,including 121 medicine cases,the total amount of transfusion was about 60.9％ (1 385/2 275),78 surgical cases accounted for 39.1％ (890/2 275);In the distribution of diseases,the blood system diseases accounted for 49.2％ (1 120/2 275),the total amount of obstetrics and gynecology disease infusion accounted for 10.6％ (240/2 275),and the amount of tumor radiotherapy and chemotherapy accounted for 6.2％ (140/2 275);The proportion of ABO blood type distribution was O type 25.9％,A type 22.9％,Btype 20.7％,ABtype 30.5％,respectively;3) The Plt counts of group A and B were significantly different before and after transfusion (P ＜0.05).But there was no significant difference between the two groups of cases before transfusion and 24h Plt count after transfusion,the Plt counts difference,and 24 h CCI (P＞0.05);4) The effective rates of platelet transfusion in group A and B were 76.9％ and 76.7％,respectively.Which has no significant difference between the two groups (P＞0.05)).There was no significant difference between the two groups in medical and surgical cases (P＞0.05),but the effective rate of surgical cases in group A (84.6％) was higher than that in B group (75.3％).The difference effect of medicine and surgery cases in B group were not statistically significant (P＞0.05),but the difference effect of medicine and surgery cases in A group was statistically significant (P＜0.05),platelet transfusion inefficient in surgical cases (15.4％) was significantly lower than that of cases (28.1％);5) The incidence of adverse reactions of blood transfusion was 3.5％,4.7％ in group A and B,and the blood transfusion mortality rate was zero,the difference was not statistically significant (P＞ 0.05).Conclusion The clinical effectiveness and safety of cryopreserved apheresis platelets are similar to those of fresh apheresis platelets,and the former can be widely Used in clinic,in particular,it has certain advantages in the surgical hemostatic effect.but for repeated infusion cases or platelet transfusion ineffective cases should be given priority to fresh apheresis platelets.

Objective To evaluate the safety and clinical efficacy of stent implantation using a rapid artificial cardiac pacing technique on ostial lesions of left anterior descending artery (LAD).Methods From Jun 2008 to Nov 2010,38 patients with ostial lesions of LAD were recruited and randomly divided into two groups:patients with stent implantation using a rapid artificial cardiac pacing technique (pacing group,n =19 ) and patients with direct stent implantation (no-pacing group,n =19 ).Post-stenting examination was performed.Patients were followed-up for 9 months and coronary angiography was reviewed.The immediate success rate,major adverse cardiac events including death,reinfarction and target vessel revascularization,late lumen loss,sent thmmbosisin,rent-restenosis were compared between these two groups.Results There were no significant differences in the baseline values,disease characteristics and instant response to surgery between pacing and no-pacing groups(P ＞ 0.05 ).The time cost for stent placement was significantly shorter in the pacing group than that in the no-pacing group ( [ 16.5 ± 0.5 ] s vs.[46.6 ± 1.4 ] s,t =88.256,P =0.004 ).After surgery,there was no acute or subacute thrombosis,in-stent restenosis or occlusion for patients in the pacing group.In the no-pacing group,one patient developed acute thrombosis.The symptoms disappeared after thrombus aspiration and balloon dilatation by emergency percutaneous coronary intervention.Patients were followed up for 270 -275 days,and patients in the pacing group received post-stenting coronary angiography 9 months after stent implantation using a rapid artificial cardiac pacing technique,and no in-stent re-stenosis was found.During this period,nobody had adverse events such as death,myocardial infarction or target lesion revascularization,while five cases with in-stent restenosis (50％ -60％ )were found in the no-pacing group,without further target lesion reconstruction due to symptomless.Conclusion Compared with previous positioning technique,stent implantation using a rapid artificial cardiac pacing technique on treatment of ostial lesions of LAD is safer,and more effective.It is a favorable method for accurate positioning of bracket and can improve the prognosis,reduce the occurrence of acute thrombosis and in-stent restenosis.

AIM: To study the effect of Jiunaoning Injection on apoptosis and the changes in cytosolic calcium induced by hypoxia/hypoglycemia in SH-SY5Y human neuroblastoma cells. METHODS: The apoptosis and the changes in cytosolic calcium were induced by hypoxia and hypoglycemia .Apoptosis was measured via flow cytometry(FCM), and cytosolic calcium was determined by Fluo-3 fluorometry. RESULTS: The hypoxia/hypoglycemia induced apoptosis and increased the cytosolic calcium in SH-SY5Y cells. Jiunaoning Injection inhibited the apoptosis of the SH-SY5Y cells and decreased the concentration of cytosolic free calcium. CONCLUSION: Hypoxia/hypoglycemia induced apoptosis and increase in cytosolic free calcium in SH-SY5Y cells, which were significantly inhibited by Jiunaoning Injection.