Objective To compare the effect of different general anesthesia method on preoperation cognitive function in elderly patients.Methods According to the digital table,82 elderly patients(non cardiac surgical operation) were selected and divided into the two groups,each group 41 cases.The group A was given to sufentanil combined with propofol intravenous injection anesthesia,and the group B was given to sevoflurane inhalation anesthesia.The cognitive function of patients of two groups in preoperative,postoperative 3h,6h,24h were assessed by simple intelligence scale(MMSE),and the indicators of postoperative recovery were evaluated.Results The postoperative sleep time [(6.9 ± 1.2) min],language ability recovery time [(9.8 ± 2.2) min] in the group A were all higher than those of group B (t =6.687,5.640,all P ＜ 0.01).The MMSE scores in group A after 3h [(17.1 ± 2.2) points],6h [(19.6 ±2.7)points],24h[(22.4 ±2.3) points] were lower than those of group B(t =10.026,6.229,5.082,all P＜0.01).And the PCOD rate in group A after 24h was 36.6％,which was higher than 17.1％ in group B(x2 =3.98,P ＜ 0.05).Conclusion The inhalational anesthetic methods has more quickly postoperative recovery,lower incidence of cognitive dysfunction,better clinical effects.

OBJECTIVE:To evaluate the role of PDCA cycle management on standardizing clinical use of atomization inhala-tion drug. METHODS:Using a retrospective method,300 medical records of atomization inhalation drug use were collected from our hospital during Jan.-Dec. in 2013(non-intervention group),200 medical records collected during Apr.-Jun. in 2014(one cycle intervention group),and 180 medical records collected during Oct.-Dec. in 2014(two rounds of cycle intervention group). The use of atomization inhalation drugs was compared before and after the application of PDCA cycle management. Before and after two cy-cle intervention,4 types of medical staff were investigated on cognition of the related knowledge of atomization inhalation drugs, as physicians,pharmacists,nurses and mechanic. RESULTS:After two rounds of PDCA cycle management and intervention,the proportion of unsuitable route of administration of antibiotics,long acting glucocorticoid,non-atomization inhalation dosage form of resolving phlegm drugs,protease decreased from 54.0%,62.0%,59.7%,44.7% before intervention to 0.6%,1.1%,15.0%, 1.1% after intervention;those phenomena had not been found,such as unsuitable route of administration of TCM injection and non-atomization dosage form of theophyllines,unsuitable atomization frequence,unsuitable indication,drug mixture for preventing and controlling symptom,etc. The proportion of medical staffs being familiar with medical device,drug classification,drug selec-tion for prerention and treatment and the proportion of hospital drug stock to 93.4%,86.7%,92.4%,96.2% after intervention from 38.0%,79.0%,49.0%,39.5%before intervention. CONCLUSIONS:The application of PDCA cycle management can effec-tively regulate the use of atomization inhalation drugs in our hospital;the method can be promoted and applied.

In the process of teaching international students laboratory diagnostics,teaching mode has been actively explored. The management of teaching,the foundation of teaching team,the selection of teaching materials and reformation of teaching mode are the key points that affect the teaching quality directly.

By participating in the national medical colleges and universities students' clinical skill competition and summarizing the experiences after the competition,we reflected on problems and weakness in the past clinical teaching including weak sterile concept and lack of clinical thinking,humanities,communication skills,teamwork awareness,etc.We should take methods to future improve medical students' clinical training capabilities including strengthening the concept of sterile and clinical skills,promoting training of comprehensive clinical thinking ability and problem-solving ability,emphasizing on humane care and communication between doctors and patients; cultivating teamwork awareness thus to comprehensively enhance the overall quality of medical students.

Objective To explore the role of intracellular reactive oxygen species (ROS) in the senescence of bone marrow mesenchymal stem cells(BMSCs)in patients with systemic lupus erythematosus(SLE) and the underlying mechanisms that controls the intracellular ROS levels in vitro. Methods Human bone marrow aspirates were collected from iliac of eight donors and eight SLE patients and cultured in vitro.Morphological appearance of BMSCs at different passages was examined by inverted microscope. Nuclear size was measured by fluorescence microscope. BMSCs were monitored using the senescence associated β-galacto-sidase (SAβ-gal) assay to characterize senescence in vitro. The quantification of intracellular ROS production was detected by flow cytometry. Real-time PCR technique was used to determine the gene expressions of PI3K, KRas, NRas and FoxO3 at transcription level. The expression of FoxO3, phospho-FoxO3 (p-FoxO3),AKT and phospho-AKT (p-AKT) protein were determined by Western blot analysis. Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results There were no differences in morphology and nuclear size[(31.4±4.5) vs (28.2±4.8) μm, P=0.628] of BMSCs between SLE patients and normal controls.The percentage of SA β-gal positive BMSCs from SLE patients was higher than that from healthy controls [(31.8±9.0)% vs (12.4±0.7)%, P＜0.05]. Intracellular ROS levels of BMSCs from SLE patients increased more significantly than healthy donors in vitro (34600±9600 vs 17 958±5400, P＜0.05). No significant differences in the expression of PI3K, NRas, KRas and FoxO3 from SLE subjects were observed at mRNA levels compared with normal controls, though all showed a similar upward trend. The expression of p-FoxO3 and p-AKT of BMSCs from SLE patients increased significantly compared with healthy controls at protein levels.Conclusion These data suggest that BMSCs from SLE patients aged more quickly, with high SA β-gal activity and up-regulation of intracellular ROS, which is associated with up-regulation of p-FoxO3 and pAKT at protein levels. These results indicate that bone marrow mesenchymal cell senescence may be associated with the pathogcnesis of SLE by maintaining the lifespan of BMSCs.

Objective To evaluate the clinical significance of high frequency ultrasound in the diagnosis of duodenal obstruction in neonates.Methods Ultrasonography,clinical data and etiological diagnoses of the operation in 113 neonates with duodenal obstruction were reviewed retrospectively.The digestive tract,including stomach,duodenum,jejunoileum and colon,were examined in all patients with 8-12 MHz linear transducer before operation.Results In the 113 neonates with duodenal obstruction,63 cases were diagnosed intestinal malrotation,31 cases duodenal stenosis,14 cases annular pancreas,and 5 cases duodenal atresia.One hundred and six cases were diagnosed as duodenal obstruction by ultrasound,of which intestinal malrotation in 61 cases,duodenal stenosis in 29 cases,duodenal atresia in 4 cases,and annular pancreas in 12 cases.The diagnostic rate was 93.81% (106/113 cases),96.83% (61/63 cases),93.55% (29/31 cases),80.00% (4/5cases) and 85.71% (12/14 cases),respectively.The location of obstruction diagnosed by ultrasound was coincident with the operation in 92 cases,with a diagnostic rate of 81.42%(92/113).Conclusions High frequency ultrasound plays an important role in diagnosing the causes and location of duodenal obstruction.It can be used as the first choice of examinal methods for the neonates with duodenal obstruction.

Objective To explore the specificity and efficiency of YFP labeled natural killer (NK) cells through Vav-Cre induced YFP reporter system in mice. Methods ROSA26R-YFP and Vav-Cre mice were crossed, and their YFP and Cre gene double positive progeny were screened by genotyping. The specificity of YFP in hematopoietic cells from im-mune organs including lymph nodes, spleen, thymus and bone marrow were analyzed by flow cytometry. The percentages of YFP positive cells in NK cells from lymph nodes, spleen and bone marrow were also analyzed by flow cytometry. Results A total of 11 double positive mice (ROSA26R-YFP-(+/-)VavCre) were obtained in 17 mouse offspring by crossing ROSA26R-YFP mice with Vav-Cre mice. The percentages of YFP positive cells in immune organs including lymph nodes, spleen, thy-mus and bone marrow were 73.87%± 1.51%, 56.07%± 1.47%, 86.17%± 1.74%and 53.60%± 3.56%, and there were signifi-cant differences compared with the corresponding negative control cells(0.27%±0.01%, 1.33%±0.91%, 0.11%±0.01%and 0.29%± 0.03%, P<0.01). There were no YFP expressions in non-immune organs in double positive mice and in negative control mice (0.72%±0.43%vs 0.92%±0.27%, P>0.05). The positive rates of YFP were significantly higher in NK cells in lymph nodes, spleen and bone marrow (76.94%±0.84%、81.66%±1.18%and 88.92%±0.77%) compared with those of control (P<0.01). Conclusion YFP marked NK cells through Vav-Cre induced YFP reporter system in mice have high specificity and efficiency.

Background and purpose:The human oncogene B-cell-speciifc moloney murine leukemia virus integration site 1 (Bmi-1) is an important member of the polycomb group family, and it regulates cell proliferation and senescence via INK4a/ARF locus. This study investigated the effects of Bmi-1-siRNA on the proliferation of lung adenocarcinoma cell line SPC-A1 cells with INK4a/ARF locus and clarify the mechanism of Bmi-1-mediated effect on proliferation of lung adenocarcinoma cells. Methods:In this study, we chose the most efifcient siRNA chain the pGeneshl-2-Bmi-1 sense-1 and inserted into a pSUPER-retro-neo retroviral vector. The packaged si-Bmi-1 pSUPERret-ro-neo retroviral vector was stably transfected into lung adenocarcinoma SPC-A1 cell line. The stably transfected cells were cultured and passed. After transfection, the levels of Bmi-1 mRNA and protein expression of SPC-A1 cells were analyzed by RT-PCR and Western blot respectively. Trypan blue, MTT and plate colony forming assay were performed to observe the proliferation capibility of SPC-A1 cells and evaluate the cloning forming ability in vitro. The potency of tumorigenesis was observed in nude mouse through hypodermic inoculation of SPC-A1 cells. Cell cycle distribu-tion was analyzed by lfow cytometry (FCM) in SPC-A1 cells. The expression levels of proliferation proteins including p16INK4a, p53, Cyclin D1, PTEN, Akt and Ser473p-Akt were analyzed by Western blot. Results:The mRNA and protein expression levels of Bmi-1 were signiifcantly reduced in SPC-A1-Bmi-1-siRNA cells transfected with pSUPER-retro-neo retroviral vector. Knockdown of Bmi-1 could inhibit the growth, colony formation in vitro and tumorigenesis in vivo of SPC-A1 cells (P<0.01). The transfected SPC-A1 cells were arrested in G1 phase [(64.6±1.2)%, P<0.05]. Compared with two control groups, p16INK4a, p53 and Akt were not affected (P>0.05), while Cyclin D1 and Ser473p-Akt were downregulated (P<0.01) and PTEN was up-regulated (P<0.01) in the SPC-A1-Bmi-1-siRNA cells. SPC-A1-Bmi-1-siRNA cells were treated with various concentrations of PTEN inhibitor to determine expression levels of PTEN, Bmi-1 and Ser473p-Akt protein. Ablation of PTEN rescued Bmi-1 and Ser473p-Akt expression in SPC-A1-Bmi-1-siRNA cells. Conclusion:Knockdown of Bmi-1 gene can arrest the proliferation of SPC-A1 cells through G0/G1 phase arrest by inhibiting Cyclin D1 expression indirectly, which may be not associated with p16INK4a signaling pathway.

Background and purpose:The pro-oncogene Bmi-1 is a member of the polycomb-group family, can regulation of the proliferation and self-renewal of normal and tumor stem cells. In recent years, Bmi-1 has been found that it is overexpressed in varieties of human malignant tumors. The study aimed to observe the effects of Bmi-1-siRNA on the growth capacity of lung cancer cell line A549 in vivo and in vivo, and explore its mechanism. Methods:The most effective one as a target sequence was chosen from four Bmi-1 siRNA sequences which were designed by our lab, and one random sequence was chosen as a negative control. In short, the chemically synthesized siRNA and control sequences were connected to a retrovirus expressing vector, pSUPERretro-Neo plasmid, and then transfected into A549 cells. The stably transfected cells were cultured and passed. The level of mRNA and protein of Bmi-1 in A549 cells were assessed by RT-PCR and Western blot respectively. The proliferations of A549 cells in vivo was analyzed with MTT, trypan blue exclusion and plate colony forming methods. Flow cytometry was used for cell cycle analysis. The potency of tumorigenesis was observed in nude mouse through hypodermic inoculation of A549 cells. The expressions of cyclin D, p21/27, p-AKT and PTEN were analyzed by Western blot. Results:Compared to A549-ctr and A549-wt cells, Bmi-1 mRNA and protein levels all signiifcantly reduced in A549-Bmi-1-siRNA cells. Bmi-1-siRNA inhibited the growth, colony formation in vitro and tumorigenesis in vitro of A549 cells, and the interference cells cell cycle arrested in G1 phase. In A549-Bmi-1-siRNA cells, p-AKT and cyclinD1 expression were down-regulated while p21/p27 and PTEN were up-regulated. Conclusion:Silencing Bmi-1 gene inhibits the proliferation of A549 cells through G1 phase arrest, which involves the downregulation of cyclin D/p-AKT and upregulation of p21/p27/PTEN.

A total of 118 patients with type 2 diabetes mellitus were divided into acarbose treatment group ( A group,n =58 ) and no acarbose treatment group ( B group,n =60),and 57 healthy subjects were used as control group (C group).The quantification of fecal bifidobacteria and enterococcus faecalis in these subjects was made by realtime PCR.The results showed that fecal bifidobacteria contents in A and B groups were lower and enterococcus faecalis contents were higher compared with C group.After four weeks of intervention,fecal bifidobacteria contents in A and B groups increased ( P＜0.01 ),especially in A group,while enterococcus faecalis contents decreased ( P＜0.05 )compared with the baseline.Univariate correlation analysis showed that bifidobacteria content was negatively associated with lipopolysaccharides(LPS),advanced glycation index,high sensitive C reactive protein ( hs-CRP),and body mass index ( BMI ) at baseline ( P＜0.05 or P＜0.01 ).The enterococcus faecalis content was positively associated with levels of monocyte chemoattractant protein-1,LPS,tumor necrosis factor-α,hs-CRP,plasminogen activator inhibitor-1,BMI,and HbA1c (P ＜0.01 ).After four weeks of intervention,the above associations disappeared.Stepwise multivariate regression showed that basal BMI,HbA1c,and age contributed to the increase in the number of enterococcus faecalis,and BMI negatively contributed to the decrease in number of bifidobacteria.