Objective To investigate the vicissitude of infection pathogens and their change in resistance to antibiotics in our hospital in the past 10 years, and to offer scientific information for clinical anti-infection treatment. Methods The data of pathogens identified and susceptibility test with VITEK system as well as K-B methods from 1995 to 2002 were analyzed. Results Gram negative bacteria was the predominance bacteria in recent 5 years, accounting for 60.7%-70.2% of all pathogens, with Pseudomonas aeruginosa ranked first, followed by Escherichia coli, Klebsiella pneumoniae, Serratia sp, Enterobacter sp and Acinetobacter sp. The ratio of Acinetobacter sp seemed to be increased in 1998, and it kept a high level in recent years. The isolation rate of Staphylococcus aureus from clinical specimens was the highest among pathogens since 1999, and 42.9%-74.5% of them were Methicillin-resistance S. aureus (MRSA). MRSA was found to be highly resistant to many antibiotics, and there was a tendency of increasing resistance to all kinds of antibiotics in Ps. aeruginosa and Acinetobacter species. Conclusion The significant changes in infectious pathogens in our hospital were an increase in S. aureus and decrease in E.coli in constituent ratios, as well as an elevation of drug resistance level of predominate bacteria. The results suggest that corresponding adjustment should be made in the strategy of infection treatment.

Objective To study the mechanism of resistance to antibiotics and disinfectants of Pseudomonas aeruginosa isolated from clinical specimens in a hospital, and to provide a reference for the use of aminoglycoside antibiotics for the treatment of P. aeruginosa infection as well as disinfection and sterilization. Methods 35 strains of multi-resistant P. aeruginosa were screened from clinical specimens by susceptibility test of agar dilution. Five kinds of aminoglycoside modifying enzyme gene and qzcE?1 gene were detected by polymerase chain reaction (PCR) method. Results The positive rates of aminoglycoside modifying enzyme genes, including aac(3)-Ⅱ, aac(6′)-Ⅰ, aac(6′)-Ⅱ, ant(3″)-Ⅰ and ant(2″)-Ⅰ, were 48.6%, 40%, 54.3%, 45.7% and 60%, respectively, and nearly all strains were positive for 2 or more than 2 kinds of above aminoglycoside modifying enzyme gene. The positive rate of qzcE?1 gene was 94.3%. Conclusions There was a close relationship between aminoglycoside modifying enzyme producing P. aeruginosa and its multi-resistance to antibiotics, The results suugested that aminoglycosides should be used cautiously, and it should be based on the result of susceptibility test in the treatment of P. aeruginosa infection, and it was inadvisable to use quaternary ammonium and biguanides disinfectant in disinfection and sterilization.

Objective To observe the serum levels of homocysteine (Hcy) and hypersensitivity C-reactive protein(hs-CRP) in elderly patients with cerebral infarction and to investigate their relationship and clinical significance by National Institutes of Health Stroke Scale (NIHSS).Methods The serum levels of Hcy and hs-CRP were by enzymatic cycling method and scattering turbidimetry in the elderly patients ( 116 cases with cerebral infarction and 100 cases of healthy control).Those 116 cases with cerebral infarction were divided into three groups by the degree of NIHSS.The three groups were compared with each other.Results The levels of serum Hcy and hs-CRP in elderly patients with cerebral infarction were significantly higher than that of healthy control group ( t =6.97,P ＜0.01 ; t =14.96,P ＜0.01 ).There has significant difference among those three groups with cerebral infarction by comparing with each other( F =23.49,P ＜0.05; F =28.19,P ＜0.05).A positive correlation was found between Hcy and degree of NIHSS( r=0.54,P ＜0.05),and between hs-CRPand degree of NIHSS( r =0.58,P ＜0.05).Conclusions Serum levels of Hcy and hs-CRP are correlated with the occurrence of cerebral infarction and its severity.There has positive clinical significance to evaluate the effect of cerebral infarction by measuring the serum levels of Hcy and hs-CRP dynamic.

Objective To study the role of miR-320-3p in adipocyte differentiation. Methods Marrow mesenchymal stem cells were isolated from mice and cultured then induced with adipogenic agents for 3 days. The transcription level of miR-320-3p was examined by qRT-PCR. Stromal ST2 cells were transfected with miR-320-3p, followed by adipogenic treatment. Oil-red O staining and qRT-PCR were employed to assess the differentiation of adipocytes induced by miR-320-3p. Results The expression level of miR-320-3p increased in MSCs after adipogenic treatment (P < 0.01). Addition of miR-320-3p in ST2 cells promoted the formation of oil-red O positive adipocytes and up-regulated the expression levels of adipogenic transcription factors such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α(C/EBPα) and the marker gene adipocyte fatty acid binding protein 4 (FABP4),compared to cells that transfected with miR-320-3p mimics (P<0.05). Conclusion miR-320-3p promotes ST2 cells differentiation into adipocytes.

Background and purpose:Neuroendocrine neoplasm is one kind of infrequent neoplasms from neuroendocrine cell and peptidergic neurons. This study aimed to investigate the epidemiology and clinicopathological characteristics of neuroendocrine neoplasms (NEN) in Dalian. Methods: We retrospectively analyzed all neuroendocrine neoplasms patients in First Afifliated Hospital of Dalian Medical University from 2000 to 2013. The epidemiology characteristics, pathogenic sites and pathological types were reviewed, and the differences between gender and age were also analyzed. The statistics analysis such as independent-samples t test and one-way ANOVA and chi-square test were performed. Results:There were 279 NEN cases, including 166 male and 113 female patients (male:female=1.14:1), detection rate was 0.258%. The mean age of all cases was 59.4±17.1 (20-83), for male 58.9±19.4 (20-81) and female 61.7±15.0 (29-83). There was no statistical signiifcance of detection rate and mean age between male and female patients. Digestive system was the most common occurred site of NEN (71.68%), then was the respiratory system (20.79%). There was no statistical significance of pathogenic sites between male and female.In all case, 132 with neuroendocrine tumor (47.31%), 140 with neuroendocrine cancer (50.18%), 7 with mixed adenoneuroendocrine carcinoma (2.51%). There was no statistical signiifcance of pathological types between male and female (P>0.05).Conclusion:Recently, the detection rate of neuroendocrine neoplasms was raised in Dalian, but there was no signiifcance differences in detection rate,pathogenic sites and pathological types between male and female.

Objective To explore the possible risk factors that influence bone mineral density (BMD) in elderly men with type 2 diabetes. Methods A total of 80 elderly men with type 2 diabetes were included in this study. The BMD of lumbar vertebrae and femur were measured by dual-energy X-ray absorptiometry (DXA). And fasting blood and urine samples were taken to check the biochemical levels of bone metabolism and blood glucose. The correlations between BMD and other related factors were analyzed. Results In this group, the prevalence rates of osteoporosis and osteopenia were 18. 6% and 53.8%, respectively. Body mass and body mass index (BMI) were positively correlated with BMD at all sites (r=0. 202～0. 298, P<0. 05 or P<0. 01). However, age and glycosylated hemoglobin (HbAlc) were negatively correlated with BMD of lumbar and femoral neck (r=-0. 172～-0. 211 ,all P<0. 05). Leptin was not only positively correlated with the BMD of femoral neck and Ward's triangle, but also with body mass, BMI, fasting blood glucose (FBG), total cholesterol and HbAlc (r=0. 219 ～ 0. 509, P<0.05 or P<0.01). Using stepwise regression analysis, body mass was the predictor of BMD at all sites assessed, while the HbAlc and leptin levels could respectively influence BMD at femoral neck and Ward's triangle (r~2= 0. 196 ～ 0. 276, all P< 0. 01). Conclusions It may suggest that differential factors predict the variance of BMD at different sites in elderly men with type 2 diabetes.

Adolescent ; Adult ; China ; epidemiology ; Humans ; Middle Aged ; Motor Activity ; Rural Population ; Suburban Population ; Urban Population

Objective To explore the impact of calciotropic hormones,such as parathyroid hormone (PTH)and vitamin D,on bone mineral density(BMD)in the old men with type 2-Diabetes.Methods Sixty elderly men with type 2-Diabetes were submitted to dual-energy X-ray absorptiometry to evaluate the BMD at lumbar spine and hip.Fasting blood samples were collected to evaluate the indexes of bone metablism and blood glucose.PTH and 25-(OH)-Vitamin D3 were measured and analyzed for their correlation with BMD at different sites.Results In all patients,the percentage of osteoporosis and osteopenia accounted for 20.0％ and 53.3％ of the patients according to BMD at lumbar or hip.Compared with the patient group with normal BMD,serum PTH was significantly higher in the patient group with osteopenia or osteoporsis([44.87 ± 10.62]μg/L vs[36.96 ±12.36]μg/L,P ＜ 0.05 ;[50.24 ± 20.32]μg/L vs[36.96 ± 12.36]μg/L,P ＜ 0.05).But there was no difference in 25-(OH)-Vitamin D3 levels between all groups.PTH was correlated negatively with BMD at hip (r =-0.224,P ＜ 0.05),but not significantly correlated with BMD at lumbar(r =-0.187,P ＞ 0.05)Conclusions Serum PTH was correlated negatively with BMD at hip in elderly man with type 2-Diabetes.

Objective Our hypothesis was if we rendered host' s cytotoxic tumor lymphocytes insensitive to TGF-β,these immune cells could be able to overcome the TGF-β mediated immunosuppression and reject the tumor.We aimed to develop a lentivirus mediated gene transfer program incorporating a herpes simplex virus thymidine kinase(HSV-tk)in our dominant negative TGF-β type Ⅱ receptor(TβRⅡDNglytk)expression vector.So we first need to construct the lentiviral pLenti6/V5-D-TOPO vector containing TβRⅡDNglytk and produce the recombinant lentivirus as the transfection vector.Methods PCR were used to amplify the genes TβRⅡDN and HSV-tk from the respective plasmids.Then the genes were linked by recombinant PCR technology to construct the fusion gene TβRⅡDNglytk and control vector TRANSglytk.According to the operation manual from the Invitrogen company,TOPO cloning technology was used to construct the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk.Both of the constructed plasmids were verified by sequencing.ViraPowerTM Lentiviral System and 293 FT cells provided by Invitrogen were used to produce the recombinant lentivirus vector,the tillers of the lentivirus were determined by 293 cells.Results The construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk were completed succefully.The DNA sequencing results showed that both the plasmids were constructed correctly.Using them we successfully produced infectious lentivirus vectors with appropriate tilters.Conclusions Using TOPO cloning technology in the construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk and recombinant PCR in the fusion genes is feasible.Recombinant PCR combine with TOPO cloning technology can be a simple,highly efficient and rapid way to construct lentiviral vector.The production of infectious lentivirus with appropriate tilters using 293FT is suitable and feasible.The construction of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and production of the infectious lentivirus will lay a foundation for immunotherapy of prostate cancer.

Objective: To construct the lentiviral plenti6/V5-D-TOPO vector containing TβR Ⅱ Dnglytk and control vector by developing a lentivirus mediated gene transfer program incorporating a herpes simplex virus thymidine kinase(HSVtk) in the dominant negative TGF-β type Ⅱ receptor (TβR Ⅱ Dnglytk) expression vector. Methods: The PCR methods were used to amplify the genes TβR Ⅱ DN and HSV-tk from the respective plasmids. Then the genes were Linked by recombinant PCR technology to construct the fusion gene TβR Ⅱ Dnglytk and control vector TRANSglytk. According to the operation manual (from the Invitrogen company), ToPe cloning technology were used to construct the plasmids of plenti6/VS-D-TOPO(R)-TβRⅡ Dnglytk and plenti6/VS-D-TOPO -TRANSglytk. Both of the constructed plasmids were verified by sequencing.Results: The constructions of the plasmids of plenti6/V5-D-TOPO(R)-TβR Ⅱ Dnglytk and plenti6/V5-D-TOPO(R)-TRANSglytk were completed smoothly. The DNA sequencing results showed that both the plasmids were constructed correcdy and can be used in the production of infectious lentivirus vectors. Conclusion: Using TOPO cloning technology in the construction of the plasmids of plenti6/VS-D-TOPO -TβR Ⅱ Dnglytk and plenti6/VS-D-TOPO(R)-TRANSglytk and recombinant PCR in the fusion genes are feasible. The recombinant PCR combined with ToPe cloning technology can be the simple, highly efficient and rapid way to construct lentiviral vector and construction of plenti6/V5-D-TOPO(R)-TβR Ⅱ Dnglytk, which will lay a foundation for tumor immunotherapy.