Objective To analyze the related factors of lymph node metastasis in N2 group of patients with non-small cell lung cancer,and found its meaning for lymph node dissection.Methods A retrospective analysis was made on 110 patients with non-small cell lung cancer from July 2014 to May 2016 in our thoracic surgery department.Potential related factors were collected,single factor analysis and variate analysis were carried out to find the relationship between N2 lymph node metastasis and potential related factors.Results Univariate analysis showed that the longest diameter of the tumor ＞ 2 cm (P =0.016),lymph node imaging (P =0.021),pleural involvement (P =0.002) were related factors of lymph node metastasis in N2 group,and these three factors were independent related factors of lymph node metastasis in N2 group.Conclusions The longest diameter of the tumor,lymph node imaging and pleural involvement are related factors of N2 lymph node metastasis.Systematic lymph node dissection is strongly recommended for patients with three related factors at the same time.

Objective To analyze the dominant genotypes of porin Ⅰ gene encoding major outer membrane protein of Neisseria gonorrhoeae in Zhejiang Province and to construct the prokaryotic expression systems of porinⅠA and porinⅠB genes.Methods Based on the previous research,the porinⅠA and porinⅠB genes were sequenced after T -A cloning, and the dominant genotypes of porinⅠA and porinⅠB genes were analyzed.Then the prokaryotic expression systems of the dominant genotypes of porinⅠA and porinⅠB genes were constructed.Ten percent SDS -PAGE was applied to measure the output of PⅠA and PⅠB proteins after inducement by 0.5 mmol/L IPTG.Results All 5 porinⅠA isolates sequenced were serovar ⅠA -6.Of the 11 porinⅠB isolates sequenced,there were 5 serovarⅠB -3 isolates,3 serovarⅠB -3 /6 isolates,1 serovarⅠ B -6 isolate and 2 mutant isolates.Outputs of P Ⅰ A and P Ⅰ B expressed by the constructed prokaryotic expression systems PET -42 -PⅠA and PET -42 -PⅠB were as high as 30% and 20% respectively. Conclusion ⅠA -6 is the dominant genotypes of porinⅠA gene and ⅠB -3 is the dominant genotypes of porinⅠB gene in Zhejiang Province.Comparing to the porinⅠB gene,porinⅠA gene is more conserved.The prokaryotic expression systems with high efficiency of porinⅠA and porinⅠB genes were successfully constructed,which may be helpful in the further research of genetic engineering vaccine and clinical detection of Neisseria gonorrhoeae.

OBJECTIVETo investigate the effect of arctiin on advanced oxidation protein product (AOPP)-induced epithelial-to-mesenchymal transition (EMT) in tubular cells and explore the mechanisms underlying this effect.

METHODSHuman proximal tubular cells (HK-2 cells) were treated with bovine serum albumin (BSA) or AOPPs in the presence or absence of arctiin. The expressions of E-cadherin, vimentin, and GRP78 at the protein and mRNA levels in the cells were examined using Western blotting and quantitative real-time PCR. The level of reactive oxygen species (ROS) was measured by flow cytometry with DCFH-DA as the fluorescent probe.

RESULTSCompared with BSA-treated cells, the cells treated with AOPPs showed decreased expression of epithelial cell marker E-cadherin and overexpression of mesenchymal marker vimentin and endoplasmic reticulum stress marker GRP78 with an increased ROS level. These changes induced by AOPPs were partly inhibited by arctiin.

CONCLUSIONArctiin can ameliorate AOPP-induced EMT in tubular cells by inhibiting endoplasmic reticulum stress, and oxidative stress response may participate in this process.

Advanced Oxidation Protein Products ; adverse effects ; Cadherins ; metabolism ; Cell Line ; Endoplasmic Reticulum Stress ; Epithelial Cells ; cytology ; drug effects ; Epithelial-Mesenchymal Transition ; Furans ; pharmacology ; Glucosides ; pharmacology ; Heat-Shock Proteins ; metabolism ; Humans ; Kidney Tubules ; cytology ; drug effects ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Vimentin ; metabolism

Based on the character of the molecular structure, the prodrugs of phosphates and phosphonates were divided into two categories. The first is the drug which contained the phosphate group, introducing protected groups to increase lipophilicity and improve bioavailability. The other one is the drug which had no phosphate group, introducing the phosphate group into molecules to enhance the solubility, regulate the distribution coefficient and enhance the drug-like property. This review focuses on the application of phosphates and phosphonates in drug research and development based on improvement of physico-chemical property, drug safety and the pharmacokinetics.
Animals ; Biological Availability ; Drug Design ; Drug Stability ; Humans ; Molecular Structure ; Organophosphonates ; chemical synthesis ; chemistry ; pharmacokinetics ; Phosphates ; chemical synthesis ; chemistry ; pharmacokinetics ; Prodrugs ; chemical synthesis ; chemistry ; classification ; pharmacokinetics ; Solubility ; Structure-Activity Relationship ; Tissue Distribution

Objective To develop a scale suitable for evaluating the scientific research ability of medical workers in Liaoning Province,analyze its reliability and validity to provide a scientific and objective measurement method for the evaluation of scientific research ability of medical workers in Liaoning Province.Methods A survey of 6 577 medical workers in 2 grade and up medical institutions of Liaoning province was carried out by completing a self-assessment scale form of research capacity,5 812 effective questionnaires were collected,335 out of which were used for exploratory factor analysis,5 477 were used for confirmatory factor analysis and reliability and validity analysis.Results The scale of Cronbach's a coefficient is 0.982,the retest reliability is 0.967,the cumulative contribution rate of the four factors extracted by exploratory factor analysis was 80.195％.The validation factor analysis showed that the model fit well (GFI=0.907,AGFI=0.881,CFI=0.962,RMESA=0.076,NFI=0.960,IFI=0.962).Conclusions The research capability scale for Liaoning medical workers has good reliability and validity,which can be used in the evaluation of the research capability of the medical workers in Liaoning province.

Objective:To apply ¹³N-ammonia PET/CT cerebral blood perfusion imaging combined with methazolamide challenge for cerebrovascular reserve (CVR) evaluation in ischemic cerebrovascular diseases. Methods:From January, 2014 to December, 2016, 56 ischemic stroke patients with serious stenosis of unilateral internal carotid artery or middle cerebral artery accepted basal and stress PET/CT with methazolamide challenge. The patients were divided into normal-CVR group (n = 29) and reduced-CVR group (n = 27) according to the results of CVR, and followed up for 24 months. The ischemic cerebrovascular events and cerebral blood flow were observed. Results:The incidence of transient ischemic attack was more in the reduced-CVR group than in the normal-CVR group (χ² = 4.389, P < 0.05), while the incidence of ischemic stroke increased a little with no significant difference between the two groups (P > 0.05). The CBF was improved in normal-CVR group after treatment (t = 2.409, P < 0.05), and the improvement was not significant in reduced-CVR group (t = 0.648, P > 0.05). Conclusion:¹³N-ammonia PET/CT cerebral blood flow perfusion imaging combined with methazolamide challenge can be used to evaluate CVR to predict the outcome for patients with cerebral ischemic disease, which is helpful for early intervention.

Progressive tumor growth is dependent on angiogenesis. The mechanisms by which endothelial cells (ECs) are incorporated to develop new blood vessels are not well understood. Recent studies reveal that the ezrin radixin moesin (ERM) family members are key regulators of cellular activities such as adhesion, morphogenetic change, and migration. We hypothesized that ezrin, one of the ERM family members, may play important roles in ECs organization during angiogenesis, and new vessels formation in preexisting tissues. To test this hypothesis, in this study, we investigated the effects of ezrin gene silencing on the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. HUVECs were transfected with plasmids with ezrin-targeting short hairpin RNA by using the lipofectamine-2000 system. Wound assay in vitro and three-dimensional culture were used to detect the migration and angiogenesis capacity of HUVECs. The morphological changes of transfected cells were observed by confocal and phase contrast microscopy. Our results demonstrated that the decreased expression of ezrin in HUVECs significantly induced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also reduced cell migration and angiogenesis capacity in vitro, suggesting that ezrin play an important role in the process of HUVECs migration and angiogenesis.
Cell Movement ; genetics ; Cytoskeletal Proteins ; genetics ; metabolism ; Cytoskeleton ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; physiology ; Humans ; Neovascularization, Physiologic ; genetics

Background: Plasminogen activator inhibitor 1 (PAI-1) was previously established to impact several phenotypes in many kinds of cancer, including pancreatic cancer. However, its prognostic significance in pancreatic ductal adenocarcinoma (PDAC) needs support of further evidence. This study was designed to address the issue. Methods: PAI-1 expression was detected by tissue microarray-based immunohistochemical staining in formalin-fixed paraffin-embedded specimens from 93 PDAC patients with surgical resection from September 2004 to December 2008. Its relationships with clinicopathologic variables and tumor-specific survival (TSS) were further evaluated using Chi-square, Kaplan-Meier, log-rank, as well as Cox regression analyses. Results: Expression of PAI-1 was much higher in tumor than that in nontumor tissues, based on comparison of all samples and 74 matched ones (95 [47.5, 180] vs. 80 [45, 95], Z = -2.439, P = 0.015 and 100 [46.9, 182.5] vs. 80 [45, 95], Z = -2.594, P = 0.009, respectively). In addition, tumoral PAI-1 expression was positively associated with N stage (22/35 for N1 vs. 21/51 for N0, χ = 3.903, P = 0.048). Univariate analyses showed that TSS of patients with high PAI-1 tumors was significantly poorer than that of those with low PAI-1 tumors (log rank value = 19.00, P < 0.0001). In multivariate Cox regression test, PAI-1 expression was identified as an independent predictor for long-term prognosis of resectable PDAC (hazard ratio = 2.559, 95% confidence interval = 1.499-4.367, P = 0.001). Conclusion These results suggest that expression of PAI-1 is upregulated in PDAC and might serve as a poor prognostic indicator.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Pancreatic Ductal ; chemistry ; mortality ; pathology ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Pancreatic Neoplasms ; chemistry ; mortality ; pathology ; Plasminogen Activator Inhibitor 1 ; analysis ; Prognosis ; Proportional Hazards Models

OBJECTIVETo explore the effect of infusing G-CSF mobilized recipient spleen cells at different time after haploidentical stem cell transplantation(HSCT) on graft-versus-host disease (GVHD) in mice and its possible mechanism.

METHODSForty mice after HSCT were randomly divided into 4 groups (n=10): GVHD positive control group (control group), 1st d recipient cell infusion group after transplantation (+1 d group), 4th d recipient cell infusion group after transplantation(+4 d group), 7th d recipient cell infusion group after transplantation(+7 d group). The mice in control group were injected the normal saline of same equivalent with experimental group which were given the same amount of G-CSF-mobilized recipient spleen cells. The general manifestation and pathological change of GVHD were observed. The expression changes of CD3CD4, CD3CD8cell subsets and FasL in peripheral blood were detected by flow cytometry.

RESULTSThe incidence of GVHD was significantly decreased in +4 d group and the median survival time was longer than 60 days, which was significantly higher than that of control group (24 d), +1 d group (21 d), +7 d group (28 d). (P<0.01, P<0.01, P<0.01). The Fasl expression of peripheral blood T lymphocytes in +4 d group were significantly lower than that in the other 3 groups(P<0.05).

CONCLUSIONThe +4 d infusion of G-CSF mobilized recipient spleen cells on 4th day after haploidentical HSC transplantation can inhibit the expression of FasL in donor T lymphocytes, and significantly reduce the incidence of GVHD.

OBJECTIVETo investigate the effects of microvesicles(MV) isolated from human peripheral blood hematopoietic stem cells(PB-HSC) on immune regulation and hematopoiesis.

METHODSPB-HSCs were separated by density-gradient centrifugation and cultrued. The supernatants of PB-HSC at 48 h were harvested for isolation and purification of MV by using ultracentrifugation. The electron microscopy was used to observe the morphology of MV. The protein level in MV was quantified through bicinchoninic acid(BCA) protein assay. Flow cytometry was used to detect the immunophenotype of MV. Human peripheral blood mononuclear cells(PB-MNC) were isolated from healthy donor and treated with isolated MV. After being co-cultured for 12 h, confocal microscopy was used to observe the action mode of MV on PB-MNC. After being co-cultured for 48 h, the levels of IL-2, IL-6, IL-8, IL-10, IFN-γ and TNF-α were detected by ELISA. Flow cytometry was used to detect the changes of T cell subsets and the activation of T cell subsets as well as intracellular cytokine staining after co-culture for 48 h. The methylcellulose was used to assess the hematopoiesis-supportive function of MV as well as co-cultured supernatants.

RESULTSThe eletron microscopy revealed that MV were elliptical membrane vesicles. The protein amount in MV ranges from 29 to 110 µg. Flow cytometry showed that MV expressed mix markers on the surface, especially highly expressed MV specific marker CD63(85.86%) and hematopoietic stem cell marker CD34(33.52%). After being co-cultured for 12 h, confocal microscopy showed that MV were merged with PB-MNC. After being co-cultured for 48 h, ELISA showed that the secretion of cytokines IL-6,IL-8, IL-10 as well as TNF-α was increased while the level of IL-2 and IFN-γ was not changed much. The results of flow cytometry showed that there was no significant change in T cell subsets and T cell activation. Staining of intracellular factor showed that IL-8 was increased significantly in CD11ccells. The colony-forming experiments revealed that MV and the co-cultured supernatants could facilitate the colony formation.

CONCLUSIONMV isolated from PB-HSC have immune-regulatery function and can prornote hematopoiesis.