Purpose:To evaluate the differences in expression of carcinoembryonic antigen mRNA (CEA mRNA) in various tissues and its use in the diagnosis of digestive tract carcinoma.Methods:There were 35 patients with digestive tract carcinoma and 12 patients with non carcinoma disease in the study. The expression of CEA mRNA in the digestive tract tissues specimens was detected with nested reverse transcriptase polymerase chain reaction(RT PCR). A contrast test was detection of carcinoembryonic antigen (CEA) in blood sepcimen with enzyme linked immunosorbent assay (ELISA). Results:The expression of CEA mRNA in cancer tissues with RT PCR was 29 (82.86%) with positive results and that of CEA in blood with ELISA was 15 (42.86%) in the patients with digestive tract carcinoma. The difference in positive results was significant ( P

Objective To evaluate in vitro antifungal activity of tacrolimus combined with itraconazole or terbinafine against Exophiala dermatitidis (E.dermatitidis).Methods The minimum inhibitory concentrations (MICs) of itraconazole and terbinafine against 12 strains of E.dermatitidis were determined using the Clinical and Laboratory Standards Institute (CLSI) broth microdilution susceptibility method (M38-A2 Document).A broth microdilution checkerboard method was used to evaluate the in vitro antifungal activity of tacrolimus combined with itraconazole or terbinafine against E.dermatitidis.Results The MIC ranges of terbinafine and itraconazole against E.dermatitidis were 0.060-0.125 mg/L and 0.5-1 mg/L,respectively.The combination of tacrolimus with terbinafine showed synergistic inhibitory effects against 5 strains of E.dermatitidis,while the combination of tacrolimus with itraconazole revealed synergistic effects against 10 strains of E.dermatitidis.No antagonism was observed in either of the two combinations.Conclusion In vitro combination of tacrolimus with itraconazole or terbinafine can enhance the antifungal activity of itraconazole or terbinafine against E.dermatitidis.

ner. Furthermore, IL-10 and TGF-β1 showed a synergic effect. Conclusion IL-10 and TGF-β1, the cytokines of regulatory T cells, suppress the formation and bone resorption function of the osteoclasts.

Objective To construct an RNAi lentiviral vector targeting rat CD86 gene and detect its effect of gene silencing in dendritic cells. Methods The effective sequence of siRNA targeting rat CD86 gene was confirmed in our previous work. DNA oligo containing short hairpin frame was synthesized and re-annealed, and then cloned into pGCL-GFP lentivind expression vector. PCR and sequencing analysis were made for verifying the positive clones. The virus packaging plasmids were transfected into 293T cells to har-vest shRNA lentivirus. After infection in dendritic cells, real-time PCR and Western blot were performed to determine the expression level of CD86 at mRNA and protein of NC( negative control virus). Results PCR and sequencing analysis revealed that shRNA plasmid was correctly constructed. Virus with a titer of 2×108 TU/ml was successfully packaged. CD86 expression in dendritic cells can be knockdown at both mRNA and protein level by virus infection as characterized by 90.6% decrease of mRNA and significant inhibition of protein compared with NC. Conclusion The recombinant lentiviral shRNA expressing vector targeting rat CD86 gene has been successfully constructed and packaged. CD86 mRNA and protein can be effectively down-regulated in dendritic cells.

Objective:To construct a RNAi lentiviral vector targeting rat CD80 gene and detect its effect of gene silencing in NRK and IEC6 cells.Methods:The effective sequence of siRNA targeting rat CD80 gene was confirmed in our previous work.Oligo-DNA fragment containing short hairpin frame was synthesized and reannealed,and then cloned into pGCSIL-GFP lentiviral expression vector.PCR and sequencing analysis were made for verifying the positive clones.The virus packaging plasmids were transfected into 293T cells to harvest shRNA lentivirus.After infection in NRK and IEC6 cells,Real-time PCR was performed to determine the expressing level of CD80.Results:PCR and sequencing revealed that shRNA plasmids was correctly constructed.Virus with a titer of 4×10~8 TU/ml was successfully packaged.CD80 expression in NRK and IEC6 cells could be knockdown by virus infection as characterized by 66.9% and 63.5% decrease of CD80 mRNA in NRK and IEC6 cells respectively,compared with negative control lentivirus.Conclusion:The recombinant lentiviral shRNA expressing vector targeting rat CD80 gene has been successfully constructed and packaged.CD80 mRNA could be down-regulated availably in NRK and IEC6 cells.

Objective To evaluate the relationship between endometriosis fertility index (EFI) and pregnancies after laparoscopic surgery in endometriosis-associated infertility.Methods From Jan.2005 to Jan.2010,medical documents of 350 infertile patients due to endometriosis undergoing laparoscopic surgery were studied retrospectively.Pregnancy outcomes were followed up by telephone.EFI was calculated by history factors,least function score and some aspects of the revised American Fertility Society (r-AFS) endometriosis stage.The cumulative pregnancy rate was calculated and compared by Kaplan-Meier survival analysis.Results Within 3 years after surgery,the cumulative pregnancy rates among patients with EFI score 8,9,10 were 62.5％,69.8％ and 81.1％,respectively.There was no significant difference in pregnancy rates among those three groups of patients ( P =0.24 ).The cumulative pregnancy rates among patients with EFI score 5,6,7 were 49.8％,43.9％ and 41.6％,respectively,which did not reach statistical difference ( P =0.83 ).The cumulative pregnancy rates of EFI score 8 - 10 was significantly higher than that of EFI score 5 -7 (71.8％ vs.44.4％,P =0.000).The patients with EFI score 0 -4 was quite small with only 33 cases,among which 15 cases were pregnant.Conclusions There is relationship between EFI and pregnancy in patients with endometriosis-associated infertility.EFI is meaningful to guide post surgical treatment.

To adapt to the new challenge of nutrition and food safety as well as to cultivate talents with capacity of innovation and practical application, Dalian Medical University starts to explore a wide range of experimental teaching system:expanding classical confirmatory experiment, offering comprehensive and designing experiment , encouraging students to participate in scientific research , building Internet teaching platform, and initiating community practice service projects. Multiple dimensional and teaching patterns contribute to strengthen basic skills, interest in scientific research, innovation consciousness and practical ability of undergraduate students, which greatly improves their consciousness of social service and responsibility and helps to cultivate the practical professional talents.

Objective:To clarify the neuroprotective effects of neural cell adhesion molecule (NCAM) derived peptide P2 on in vitro cultured neuron and ischemic stroke rat. Methods:Primary cortical neurons were extracted and cultured, and CCK-8 method was used to observe the protective effect of different concentrations of P2 on cortical neurons under oxygen-glucose deprivation (OGD) conditions.The levels of apoptosis-related proteins and extracellular signal regulated kinase 1/2 (Erk1/2) were observed by Western blot. Clean grade male SD rats were selected for animal experiments. The middle cerebral artery occlusion (MCAO) method was used to establish the rat model of cerebral ischemia/reperfusion injury. The rats with successful model were divided into sham operation group, MCAO group and MCAO+ P2 group according to the random number table, with 12 rats in each group. After operation, rats in MCAO+ P2 group were subcutaneously injected with 1 mg/kg P2 once a day until 14 days after operation, and rats in the other two groups were subcutaneously injected with 0.9% sodium chloride solution of the same volume.Beam-walking test was used to evaluate the motor function of rats.Immunofluorescence staining and Western blot were used to detect the in-situ apoptosis of neuronal cells and the expression of Erk1/2 in ischemic penumbra of rat brains, respectively. All statistical analyses were performed using SPSS 22.0.Repeated measurement ANOVA was used to evaluate the beam-walking experimental data, and one-way ANOVA were used to analyze other experimental data among multiple groups.Results:Compared with OGD group, 0.5, 1.0 and 2.0 μmol/L P2 improved the activity of neurons under OGD conditions, of which 1 μmol/L P2 had the best effect ((2.436±0.284), (1.551±0.410), P<0.05). Western blot showed that the protein levels of bax ((76.120±3.232)%, (88.965±5.208)%, P<0.05), cleaved caspase-3 ((76.736±4.306)%, (97.781±8.111)%, P<0.05) and cleaved caspase-9 ((88.833±6.581)%, (104.962±4.788)%, P<0.05) in 1 μmol/L P2 treated group were all lower than those in OGD group, while the protein levels of bcl-2 ((56.146±3.882)%, (43.170±6.945)%, P<0.05) and phosphorylated Erk1/2 ((73.583±8.557)%, (55. 219±4.615)%, P<0.05) in 1 μmol/L P2 treated group were both higher than those in OGD group. Compared with MCAO group, on the 14th day after P2 intervention, the slip ratio of hindlimb of the paralyzed hind limbs of rats was lower ((23.438±11.540)%, (41.733±13.631)%, P<0.05), the apoptosis rate of neurons around the focus was lower ((13.144±6.485)%, (26. 699±6. 402)%, P<0.05), and the level of phosphorylated Erk1/2 protein in the brain tissues around the infarct focus was higher ((74.062±7.458)%, (53.327±7.093)%, P<0.05). Conclusion:Low doses of neural cell adhesion molecule derived peptide P2 exert neuroprotective effects on OGD neurons and ischemic stroke rats. The underlying mechanism may be related to the activation of Erk.

Objective To observe the clinical effects of cognitive behavioral therapy (CBT) combined with pulmonary rehabilitation on the treatment of patients with pneumoconiosis. Methods A total of 108 patients with pneumoconiosis were selected as the research subject using convenient sampling method. They were randomly divided into control group and CBT group, with 54 cases in each group. Patients in the control group were treated with conventional symptomatic treatment and pulmonary rehabilitation treatment for 12 weeks. While patients in the CBT group were treated with CBT treatment in addition to treatments of the control group. The therapeutic effect was analyzed. Results Before treatment, there was no statistical difference on score of Montreal Cognitive Assessment (MoCA), percentage of forced expiratory volume in one second to the predicted value (FEV₁%), forced expiratory volume in one second to force vital capacity ratio (FEV₁/FVC%), score of the Modified Medical Research Council Dyspnea Scale (mMRC), six minute walk distance (6MWD), Hospital Anxiety and Depression Scale (HADS) subscale scores of anxiety (HADS-A) and depression (HADS-D), and scores of Chronic Obstructive Pulmonary Disease Assessment Test (CAT), St. George's Respiratory Questionnaire (SGRQ), and serum levels of leptin, interleukin-6 (IL-6) and C-reactive protein (CRP) between the two groups (all P>0.05). After treatment, the score of MoCA, FEV₁%, FEV₁/FVC% and 6MWD increased (all P<0.05), while the scores of mMRC, HDS-A, HDS-D, CAT and SGRQ decreased (all P<0.05), and the levels of serum leptin, IL-6 and CRP decreased (all P<0.05) in the CBT group compared with the control group. Conclusion The combined treatment of CBT and pulmonary rehabilitation can better improve the cognitive ability, respiratory function, motor function, negative emotion and quality of life, and alleviate inflammatory response in patients with pneumoconiosis. It is of certain clinical application value.

Objective:To investigate the effect of S14G-humanin (HNG) on spatial learning and memory abilities and hippocampal neuron autophagy in amyloid precursor protein (APP)/presenolin-1 (PS1) double-transgenic mice.Methods:The APP/PSl transgenic mice were randomly divided into model group and HNG treatment group ( n=8). Another 8 C57BL/6J mice were chosen as controls; 0.5 mL double-steamed water was intraperitoneally injected into mice in the control group and model group every day, and 0.5 mL of HNG (50 μg/kg) was intraperitoneally injected into mice in the HNG treatment group every day; after 4 weeks of continuous injection, Morris water maze test was used to test the spatial learning and memory abilities of mice in each group. Western blotting and reverse transcription-PCR were used to detect the protein and mRNA expressions of ubiquitin like kinase1 (ULK1), P62, anti-microtubule associated protein 1 light chain 3 (LC3) II/LC3 I and cathepsin D in the hippocampus. Immunohistochemical staining was used to detect the deposition of amyloid β protein (Aβ) in the hippocampus of mice in each group. Results:Morris water maze test showed that the escape latency of mice in the control group, HNG treatment group and model group was statistically prolonged in turn, and the number of times traversing through the quadrant of the original platform was significantly smaller in turn ( P<0.05). Western blotting and reverse transcription-PCR showed that the ULK1 protein and mRNA expressions in the hippocampus decreased successively, P62 protein and mRNA expressions increased successively, LC3 II protein/LC3 I protein and LC3 II mRNA/ LC3 I mRNA ratio increased successively, and cathepsin D protein and mRNA expressions decreased successively in the control group, HNG treatment group and model group, with significant differences ( P<0.05). Immunohistochemical staining showed that the Aβ positive expressions in the hippocampal neurons of the control group, HNG treatment group and model group increased successively, with statistical differences ( P<0.05). Conclusion:HNG treatment can improve the spatial learning and memory abilities, which may be attributed to ameliorate autophagic network dysfunction and reduce Aβ plaques in the hippocampi of APP/PSl transgenic mice.