Objective To investigate the role of NF-KB binding element in regulation of NOD2. Methods Promoter region of NOD2 containing the NF-κB binding site was amplified by PCR from human genome DNA and correctly connected to the vector pEGFP-N3 which had been cut out promoter by restriction enzyme to obtain the GFP expression vector driven by human NOD2 gene promoter. The constructed plasmids were transiently transferred into cell line HeLa by LipofectAMINETM2000 and the GFP expression was ob- served by the inversion fluorescence microscope. The NF-κB binding site in the constructed vector pEGFP- N3-NOD2wt was deleted by the QuikChange site-directed mutagenesis kit. The recombinant plasmid mpEG- FP-N3-NOD2 was transiently transferred into cell line HeLa by LipofectAMINETM2000, and the GFP expres- sion was observed by the inversion fluorescence microscope. Results The constructed pEGFP-N3-NOD2wt plasmids and mpEGFP-N3-NOD2 were the same as the design confirmed by restriction digestion and se- quence analysis. The results of the cell transient transfection indicated that different strength of GEP ex- pressed by recombinant plasmids in HeLa cells could be observed. The GFP expression of constructed mpEGFP-N3-NOD2 was lower than that of pEGFP-N3-NOD2wt. Conclusion The GFP expression vector driven by human NOD2 gene promoter which contains the NF-κB binding site, and the site deleted plasmid were successfully constructed. The GFP expression of recombinant plasmid mpEGFP-N3-NOD2, deletion of the NF-KB binding site, was obviously weaken in HeLa. The results indicate that NF-KB binding element may play a positive role in regulation of NOD2 gene, which establishes favourable bases for further study on the mechanism of NOD2 gene expression and regulation.

Objective To explore the effects of progressive relaxation training on the nailfold microcirculation for psychological stress state population .Methods According to the mental health screening criteria ,60 cases of psychological healthy individuals were selected as the research object ,took the attention distribution test as psychological stress stimulation ,test self rating anxiety scale(SAS) and nailfold microcirculation function changes of test before and after stress .60 cases were divided into intervention group and normal control group ,30 cases in each group .The intervention group adopted progressive relaxation training intervention after stress ,while normal control group taking natural rest ,then detected the change of nailfold microcirculation .Results The SAS level and the nailfold microcirculation parameters before and after stress between the two groups were significantly different (P<0 .05);the difference of nailfold microcirculation morphological integral and the total integral between the two groups were statisti-cally significant(P<0 .01);and there were no difference on flow integral and around the loop integral between the two groups (P>0 .05) .Conclusion Progressive relaxation training can effectively improve the microcirculation state of the psychological stress a-mong people ,under certain conditions ,the effective intervention to the stress state population can influence the microcirculation .

Objective To investigate inhibitory effects of matrine (Ma) combined with vincristine (VCR), cytosine arabinoside (Ara-c), harringtonine (HRT), adriamycin (ADM), and daunorubicin (DNR) on proliferation of K562 cells. Methods MTT colorimetric assay was used to detect the inhibition rate of Ma combined with antineoplastic on K562 cells. Results The proliferation of K562 cells was inhibited by Ma at the concentration of 160 ?g/ml to 400 ?g/ml. The inhibitory rates of Ma combined respectively with VCR, Ara-c, HRT, ADM, and DNR were significantly higher than those of VCR, Ara-c, or HRT alone (P0.05). Conclusion Ma can inhibit the proliferation of K562 cells in a dose-dependent manner. The anti-proliferation effect of VCR, Ara-c, and HRT on K562 cells could be enhanced by Ma.

AIM and METHODS: To examine the relationship of glutathione S-transferase M1( GSTM 1) and T1( GSTT 1) with the occurrence of lung cancer, The case-control study was conducted among 161 lung cancer and 165 healthy controls. The genetic polymorphisms of GSTM1 and GSTT1 were detected with the method of multiplex polymerase chain reaction. Logistic regression analyses were used to assess the interaction of between different genotypes as well as between null genotypes and smoking. RESULTS: The frequences of GSTM 1 and GSTT 1 null genotypes had no obvious difference between lung cancer and healthy controls. In non-smoking subjects, the frequence of GSTM 1 null genotype was significantly different between lung cancer and healthy controls. Furthermore, GSTM 1 null genotype was significantly overrepresented in adenocarcinoma patients aged 60 or over, compared with controls.The results from interaction analyses showed although smoking and GSTM 1 deletion were associated with an increased risk of lung cancer, GSTM1 and GSTT 1 null genotypes combined with smoking did not have interaction effect on the risk of lung cancer. The risk for adnocarcinoma in the individuals at the age of 60 or over and in nonsmokers without GSTM1 gene but with GSTT1 functional genotype decreased by 48.5% and 45.3%, respectively. CONCLUSION: Our results suggest that GSTM1 deletion is an important host risk for lung cancer, and imply that GSTT1 functional genotype protects the old (aged 60 or over) and nonsmokers who are lack of GSTM1 gene from the risk of adenocarcinoma.

Aim To investigate the effect of matrine on cytoskeletonof K562 cells. Methods Micropipette aspiration technique was adoptedto investigate the viscoelasticity of K562 cells, while the different ex-pression of cytoskeletal protein gene was analyzed by DNA microar-ray. Results In matrine-treated K562 cells, the viscoelastic propertiesKI, K2 and were decreased significantly from 726 ± 215 to 432 ±67,433 ±119 to 242±31, 72±38 to 50±15 respectively, and the geneexpression of prefoldin and ezrin was much stronger than that of controlcells. Conclusion The strueture and function can be changed in ma-trine-treated K562 cells.

BACKGROUND:Studies have shown that sacral nerve-root stimulation based on anodes block technique can effectively reconstruct the bladder voiding function of the rabbits with spinal cord injury. But the corresponding technology of stimulating electrode has not been reported so far.
OBJECTIVE:To design and develop the stimulating electrodes matching with both rabbit sacral nerve roots and anodal blocking technique, to observe the ultrastructure and morphological change of rabbit sacral nerve roots which implanted in electrode stimulation for a long-term and to assess the safety of stimulating electrodes.
METHODS:Thirty New Zealand rabbits were included, 10 rabbits were randomly selected from them and sacrificed after anesthesia, and then cut the anterior roots of bilateral S 2 and S 3 immediately;after measuring the diameter under the light microscope, the sleeve type stimulation electrode matched with the diameter was made. The remaining 20 rabbits were randomly divided into control group and implantation group, with 10 rabbits in each group. In the implantation group, the stimulating electrodes were implanted into the forepart of S 2 and S 3 nerve roots after anesthesia, and then sacrificed after fed for half a year for col ecting the samples. Then ultrastructure change of sacral nerve roots with the implantation was observed.
RESULTS AND CONCLUSION:Structure of nerve cel s of sacral nerve roots remained in good condition under a light microscope after long-term implantation of the stimulating electrodes. No obvious degeneration of axons, no inflammatory infiltration and glial scar formation were observed. In the implantation group, myelins arranged closely without demyelination phenomenon, and there was no atrophy of neuronal nuclear, no nuclear sag, no increased nuclear decompression and heterochromatin in neurons under the light microscope. Immunohistochemical analysis showed, compared with the control group, there were no significant differences in the expressions of glial fibril ary acidic protein, Bax, Bcl-2 and Caspase-3 proteins of nerve roots in the implantation group. The stimulation electrode of rabbit sacral nerve root is developed successful y, that is, the implantation is simple and safe as it can be used for long-term implantation without histopathological changes and apoptosis.

Objective:To explore the effect of mind mapping combined with scenario simulation teaching on probation teaching of abdominal trauma.Methods:The probation undergraduates from Batch 2017 five-year clinical medicine of Air Force Medical University were selected as research objects and randomly divided into experimental group ( n=98) and control group ( n=92). The experimental group was taught by mind mapping combined with scenario simulation teaching method, while the control group was taught by traditional teaching mode. After the end of the course, the effect of teaching was evaluated from two aspects: theoretical evaluation and teaching satisfaction evaluation. SPSS 19.0 was used for t test and chi-square test. Results:The results of theoretical test in the experimental group were significantly better than those of the control group [(84.03±8.99) vs. (78.53±8.97)], and the difference was statistically significant ( P<0.05). Compared with the control group, the experimental group had obvious advantages in learning interest, teamwork awareness and clear thinking, and achieved good teaching effect. Conclusion:Therefore, this teaching model helps to improve the teaching quality and provides some reference experience.

Objective:To evaluate thrombo-pretreatment in AngioJet mechanical thrombectomy.Methods:68 acute DVT patients were randomized into two groups: thrombo-pretreatment with low-dose urokinase via popliteal vein before operation (experimental group)comparing with those undergoing upfront surgery.Results:After pretreatment, the immediate thrombus clearance grade in the experimental group was higher than that in the control group( Z=2.446, P=0.014), the procedure time was shorter [(289.1±57.9) s vs. (342.3±75.2) s] and less hemoglobin decreased after operation [(7.2±2.4) g/L vs. (11.4±2.1) g/L]. There was no difference in the deep vein patency and the incidence of PTS between the two groups at 3 , 12 and 24 months after operation. Conclusion:Pre-treatment with low dose urokinase before operation can improve the efficiency of PMT, effectively reduce the time of AngioJet activation and the destruction of red blood cells.

Objective: To evaluate the efficacy and safety of laparoscopic surgery in the treatment of gallstones and common bile duct stones. Methods: Eighty-seven patients with gallstones complicated with common bile duct stones who underwent concurrent laparoscopic surgery at Zhoushan Hospital from December 2015 to December 2017 were enrolled.The patients were divided into A group and B group according to the digital table.A group(38 cases) underwent laparoscopic cholecystectomy (LC) combined with laparoscopic common bile duct exploration (LCBDE), and B group(49 cases) underwent endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic sphincterotomy (EST) combined with laparoscopic cholecystectomy (LC). The curative effect of the two groups was observed.The operation time, the success rate of the operation and the rate of laparotomy were recorded in the two groups.The corresponding hospitalization time and cost were compared.The safety of the two different procedures was compared after surgery, and the complications of the two groups were recorded. Results: In A group, the average diameter of common bile duct stones was (1.02±0.25)cm, the average diameter of common bile duct diameter was (1.15±0.25)cm.In B group, the mean diameter of common bile duct stones was (0.99±0.26)cm, and the average diameter of common bile duct was (1.13±0.26) cm.The differences between the two groups were not statistically significant (t=0.513, 0.437, 0.367, P=2.083, 1.533, 1.095). The successful operation rate of A group was 92.11%(35/38), which in B group was 91.84%(45/49), the difference was not statistically significant between the two groups(χ²=0.006, P=0.974). The incidence rate of complications in B group was 20.41%, which was significantly higher than that in A group, the difference was statistically significant(χ²=3.654, P=0.019). The hospitalization time, hospitalization expenses in A group were (10.6±2.6)d, (26 649.8±3 478.6)CNY, respectively, which were significantly better than those in B group (t=21.971, 17.168, all P<0.05). Conclusion The efficacy of LC combined with LCBDE for patients with gallstones complicated with common bile duct stones is better than ERCP/EST combined with LC surgery, and the safety of the former is higher than the latter.

Previous studies suggest that NGAL (neutro phil gelatinase-associated lipocalin) is involved in the transformation and development of esophageal carcinoma. Alteration of NGAL expression can trigger the change of cellular morphology in esophageal carcinoma cells. However, the mechanisms remain unclear. To get a better understanding of NGAL function in esophageal carcinoma, NGAL protein was expressed in methylotrophic yeast, Pichia pastoris, and purified by chromatography. EC1.71 cells expressed high levels of NGALR (NGAL receptor) and EC109 cells expressed low levels of NGALR were used as cells model. The trafficking and the possible function of NGAL protein were then analyzed in the esophageal carcinoma cells. The results showed that 5-FAM-labeled recombinant NGAL protein could internalize into the EC1.71 and EC109 cells. Furthermore, the internalized NGAL protein could induce the alteration of cellular morphology, resulting in generation of autophagosome, transcriptional up-regulation of genes associated with autophagy and increase of phospho-ERK1/2 (p-ERK1/2). Interestingly, the treatment with the NGAL protein did not affect the intracellular iron level. These data indicate that induced autophagy by exogenous NGAL protein is a mechanism that internalized NGAL plays important roles in esophageal carcinoma cells, independent with NGAL-mediated iron transport process, while ERK1/2 signal pathway is involved in activation of autophagy by exogenous NGAL protein.