To investigate the optimum transfection condition in transfecting human keratinocytes with plasmid-liposome complexes in vitro,the cultured human keratinocytes at 60% ～ 100% confluences were exposed to the eukaryotic expression plasmid,pCMV*SPORT-β-gal,coated with LipofectAMINE in different DNA/liposome mixing concentration ratios.After cultured for another 48 hours following the ends of 6 ～ 24 hours exposures to the DNA-liposome complexes,the transfected human keratinocytes were visualized by β-galactosidase staining.Then,the transfection efficiency was determined by calculating the rate of β-galactosidase staining positive cells.β-galactosidase expression was showed clearly in human keratinocytes transfected with the DNA-liposome complexes.The highest efficiency was achieved with cultured cells at 80% and 90% confluences,demonstrating by the transfection rates of (31.35±1.35)% and (32.32±2.47)% respectively.Meanwhile,the essential transfection conditions for these efficiencies were in coating pCMV*SPORT-β-gal DNA of 1.5μg/100μL with LipofectAMINE of 12.5μL/100μL,and exposing the cells to the DNA-liposome complexes for 8 hours.These results indicate that LipofectAMINE could effectively transfer eukaryotic expression plasmid into human keratinocytes in vitro,for which the optimization of transfection conditions involve in cells growth state,DNA/liposome mixing concentration ratio,and exposure time of cells to DNA-liposome complexes.

Objective To explore the prognostic role of nutritional benefit assessment (NUTRIC score), nutritional risk screening 2002 (NRS 2002), traditional nutritional laboratory indicators albumin (ALB) and prealbumin (PA) in critically ill patients. Methods A historical-prospective cohort study was conducted. The data of 427 patients admitted to Department of Critical Care Medicine of the Affiliated Hospital of Guizhou Medical University from February 2014 to October 2014 were retrospectively analyzed, and thereafter a follow-up of 275 critically ill patients from November 2014 to April 2015 prospectively enrolled was performed. 261 patients were enrolled finally. Patients were divided into death group and survival group according to 28-day and 90-day outcome, the baseline data, acute physiology and chronic health evaluationⅡ (APACHE Ⅱ) score, sequential organ failure assessment (SOFA) score, NRS 2002, NUTRIC score, ALB and PA were compared between the two groups. Logistic regression analysis was used to find risk factors for 28-day and 90-day prognosis. Results ① NRS 2002 score of all the 261 patients were greater than or equal to 3 with 100% nutritional risk. The patients in NUTRIC score 5-9 group had lower ALB and PA, higher NRS 2002 score, longer mechanical ventilation time and length of intensive care unit (ICU) stay, which indicated they were more serious. ② Twenty eight-day mortality was 20.7% (54 died from 261). Compared with survival group, the patients in death group had higher APACHE Ⅱ, SOFA, and NUTRIC scores [29.00 (22.75, 34.25) vs. 24.00 (20.00, 28.00), 10.0 (8.0, 13.0) vs. 9.0 (7.0, 11.0), 6.37±1.84 vs. 5.59±1.64, all P < 0.01], and longer days from hospital to ICU admission and mechanical ventilation time in ICU [1.5 (0, 9.2) days vs. 0 (0, 4.0) days, 6.0 (4.0, 11.0) days vs. 4.2 (2.5, 7.8) days, both P < 0.05]. It was revealed by logistic regression analysis that APACHE Ⅱ score [odds ratio (OR) = 1.089, 95% confidence interval (95%CI) = 1.039-1.141, P = 0.000] and days from hospital to ICU admission (OR = 1.042, 95%CI = 1.014-1.071, P = 0.003) were the independent risk factors for 28-day death in critically ill patients. ③ Ninety-day mortality was 42.5% (111 died from 261). Compared with the survival group, the death group patients were older with higher APACHE Ⅱ, SOFA, NRS 2002, and NUTRIC scores [age (years): 64.44±18.11 vs. 54.25±19.66, APACHE Ⅱ: 27.00 (23.00, 31.00) vs. 23.00 (20.00, 27.00), SOFA: 10.0 (8.0, 12.0) vs. 9.0 (7.0, 11.0), NRS 2002: 5.08±1.47 vs. 4.67±1.41, NUTRIC: 6.32±1.58 vs. 5.33±1.68], ALB was significantly reduced [g/L: 27.70 (23.05, 32.00) vs. 30.73 (26.90, 34.20)], and mechanical ventilation time in ICU was extended obviously [days: 5.7 (3.6, 11.0) vs. 3.9 (2.4, 7.0), all P < 0.05]. It was revealed by logistic regression analysis that old age (OR = 1.019, 95%CI = 1.002-1.037, P = 0.029) and NUTRIC score (OR = 1.211, 95%CI = 0.983-1.491, P = 0.072) were the independent risk factors for 90-day death probability, and ALB probability was the protect factor for 90-day death (OR = 0.954, 95%CI = 0.916-0.994, P = 0.024). Conclusion It was NUTRIC score but not NRS 2002, ALB and PA predicted 90-day mortality in critically ill patients.

Aim To investigate the interaction between regulatory proteins related to the activation of eNOS and CYP4A/20-HETE.Methods The endothelia of pulmonary artery of new born bovine were cultured and divided into four groups: controlling group: ethanol solvent as vehicle (the volume is the same with 1?10~ -6 mol?L~ -1 20-HETE) was added to culture dish and incubated for 10 min; 20-HETE 5 min treatment group:1?10~ -6 mol?L~ -1 20-HETE was added to culture dish and incubated for 5 min; 20-HETE 10 min treatment group:1?10~ -6 mol?L~ -1 20-HETE was added to culture dish and incubated for 10 min; VEGF 10 min treatment group: 1?10~ -8 mol?L~ -1 VEGF was added to culture dish and incubated for 10 min. CYP4A,eNOS and proteins related to the activation of eNOS were measured by Immunoprecipitating and immunobloting.Results Expressions of eNOS and phospho-eNOS(Ser1177)in PAECs were upregulated after the cells were treated with 20-HETE, and similar phenomena were observed when 20-HETE was replaced by VEGF; 20-HETE increased the binding of Hsp、Akt and eNOS.Conclusions ①20-HETE phosphorylates eNOS at Ser1177 site which activates the eNOS to catalyze L-aginine to produce nitric oxide.②eNOS binds with Hsp90 and Akt in endothelial cell of pulmonary artery and 20-HETE increase the binding. ③eNOS binds with CYP4A in endothelial cell of pulmonary artery.

Objective To investigate the rejection mechanisms of bone grafts by comparing the immune reaction of xenografts versus allografts, and the influence of IL-4 and IL-10 on immune responses. Methods Twenty C57BL/6 mice and 1 New Zealand rabbit were used as transplant donors for allografts and xenografts, respectively. One hundred BALB/c mice were used as transplant recipients and randomly divided into 5 groups. Using a model of the muscle pouch for implantation, the immune reactions of bone allografts and bone xenografts were studied through observing the lymphocyte responses of the stimulating index of lymphocytes from the mixed lymphocyte culture (MLC), subsets of lymphocyte, cytokinetic and histological findings in 1, 2, 4 and 6 weeks after implantation respectively. Results Xenograft rejection was rapid (1 week) and stronger than allograft rejection (P

ObjectiveTo investigate the effects of bFGF on human hypertrophic scar in a nude mouse model. MethodsHuman hypertrophic scars excised in operation for burned patiens were grafted onto the back of nude mice subcutaneously and the mice were randomly divided into: control group, collagenase group, bFGF group, and bFGF plus collagenase group. Two weeks after grafting, preparations were injected into the scars for three times. The size, hardness, and morphological changes of the scars were examined. Biopsies were done 3 weeks after first injection and the histological changes were examined. ResultsIt was found that in bFGF group the size of the grafted scars reduced to 1/2～1/3 of their original size, the majority of the coarse and dense scar collagen bundles resolved and became soft loosed lump. In bFGF plus collagenase group, two thirds of the grafted scar disappeared. ConclusionTopical injection of bFGF into hypertrophic scar degrades scar collagen.

0.05). But the incidence of rehospitalization in combination group because of adverse cardiovascular events wasn’t more two times than that of non-combination group (OR=2.07; 95% confidence interval ranged from 1.87 to 2.20). CONCLUSION: The results of study are similar to other studies of foreign countries. PPI can influent the anti-platelet effect of clopidogrel so as to increase the incidence of rehospitalization risk resulted from adverse cardiovascular events. Physicians should apply PPI with cautions.

0.05) but by IL 4 the result was significant(P0.05). Conclusion CTLA4 Ig could remarkably inhibit lymphocyte proliferation in MLC than in MLBSC; and IL 4 could be more effective in MLBSC than in MLC; the result did not show any synergetic effect of CTLA4 Ig+IL 4.

To foster interdisciplinary talents with the highly fusion of clinical skills and the capacity for scientific research, a preliminary exploration of teaching for the cultivation of scientific innovation ability was carried out for clinical postgraduate students. In the cultivation of clinical post-graduate students' innovation consciousness and innovative spirit, we focused on the transformation of attaching more importance to the clinical skills than to the capacity for scientific research to establish the foundation of the competitive compound talents. Then in the early stage of medical project writing, basic knowledge learning and exchange, we stressed clinical postgraduate students' solid grasp of basic knowledge of medical science to consolidate the way of enhancing the scientific research ability. Furthermore , under the guidance of the second research supervisors allocated by the department , we strengthened the clinical postgraduate students' writing of scientific research project bid and pro-fessional paper to promote the organic combination of the clinical practice and scientific research innovation and enhance their scientific research ability.

Objective: To investigate the efficacy and safety of high-dose recombinant human interleukin-2 (rhIL-2) in treatment of patients with advanced melanoma. Methods: Twenty Chinese patients with advanced melanoma were planned to be enrolled to receive high-dose rhIL-2 treatment. The overall response rare (ORR), progression-free survival (PFS), overall survival (OS) and incidence of toxicity were evaluated. Results: Fourteen patients were enrolled, of them twelve could be evaluated. One patient achieved partial response (PR), and the maintaining duration of response was 43.0 months; three patients achieved stable disease (SD); 8 patients achieved progressive disease (PD). The median PFS was 56.0±7.6 d, and the median OS was 11.0±2.6 months. The incidence rate of grade III-IV toxicity was 40.3%, mainly manifested as gastrointestinal reaction, fever and cardiovascular reaction. Conclusion: The efficacy of high-dose rhIL-2 for Chinese patients with advanced melanoma is not as good as those previously reported in studies abroad, with high incidence rate of adverse effects. The patients can get long response duration as rhIL-2 worked. It is necessary to perform further studies on how to make choice of administration dose and select potential patients who will benefit from the treatment. Copyright© 2011 by TUMOR.

Aim To study the adaptive protection of H_2O_2 preconditioning against dopamine-induced damage in PC12 cells.Methods The apoptosis of PC12 cells was observed by electron microscope, PI stain flow cytometry (FCM) and Hoechst stain. The mitochondrial energy redox state was measured by MTT assay. The mitochondrial membrane potential(△?m) was investigated by the fluorescent probe of rhodamine 123.Results PC12 cells exposed to 50 ?mol?L -1 DA showed chromatin condensation and nucleus fragmentation observed by electron microscope. Exposure to DA (50 ?mol?L -1) for 24 h, the apoptosis of PC12 cells preconditioned by 10 ?mol?L -1 H_2O_2 for 90 min as measured by Hoechst stain was significantly decreased,compared with no-preconditioned cells. Exposure to 50,100,and 200 ?mol?L -1 H_2O_2 for 24 h, the proportion of apoptosis of PC12 cells was decreased from (20.9?1.8)%, (40.5?6.4)% and (88.1?3.9)% to (4.9?2.9)%, (12.0?1.4)%, (61.5?3.4)% after H_2O_2 preconditioning, respectively. By exposuring PC12 cells to 20,40,and 80 ?mol?L -1 DA for 24 h, the rates of MTT metabolism were reduced and the effect was prevented by H_2O_2 preconditioning. After 50 ?mol?L -1 DA exposure for 24 h,the mean fluorescence intensity of rhodamine 123 in no-preconditioned PC12 cells was decreased from (46.87?0.33) to (4.39?2.93),and that of preconditioned PC12 cells was decreased from (46.87?0.33) to (10.50?0.28). Conclusion H_2O_2 preconditioning possesses adaptive protection against dopamine-induced damage in PC12 cells.