Objective: To study the relationship between mental health and achievement motivation of undergraduates. Method: 532 undergraduates completed SCL-90 and Academic Achievement Motive Scale. Results:17.29% subjects had mental problems. Undergraduates had higher score in self-oriented motive than society-oriented motive, while the score of SCL-90 correlated positively with score in society-oriented motive. In students with poor academic achievement, the score of SCL-90 negatively correlated with score in self-oriented motive. Conclusion:Society-oriented motive had negative influence on mental health of undergraduates and in those with poor academic achievement, poor mental health positively correlated with lower self-oriented motive.

Objective To study the immunoregulatory ability of mesenchymal stem cell (MSC) on T cells. Methods MSC were isolated and cultured from bone marrow and CD3 + T cells from peripheral blood. Then the action of MSC on T cell proliferation was investigated. ELISA assay tested IFN-? and IL-10 secretion of CD3 + T cells. Results The data showed that MSC inhibited T cell proliferation in a dose dependent manner when there was 2?10 5/well (96-well plate) CD3 +T cells. While MSC stimulated T cell proliferation when there was 5?10 4/well CD3 + T cells. High dose CD3 +T cells produced high level IFN-? and IL-10 in the absence of MSC, while when there were MSC in the culture, 5?10 4/well CD3 +T cells secreted more IFN-? and IL-10 than 2?10 5/well CD3 + T cells. Conclusion The result suggested that the effect of MSC on T cells was complex, not only correlating with the cell concentration of MSC but also of T cells.

BACKGROUND: There are several types of endogenons stem/progenitor cells in lung that develop from either endoderm or mesoderm precursors.To elucidate the characteristics of the intrapulmonary stem ceils wig help to understand their biological behavior in lung injury.OBJECTIVE: To isolate the mesenchymal stem cells (MSCs) from mouse lung tissues,and identify their morphology and growth characteristics,cell surface antigens,differentiation potential in vitro,stem cell properties,and to investigate the protective role in bleomycin challenged lung.DESIGN,TIME AND SETTING: The present randomized controlled in vivo animal experiment based on in vitro observation of cytology was performed at the Laboratory of Cell Biology,Institute of Basic Medical Science,the Academy of Military Medical Science between October 2005 and August 2007.MATERIALS: Male(3-4 weeks) and female C57BL/6 (6-8 weeks) mice were used.Twenty female C57BL/6 mice were randomly and evenly divided into a pulmonary MSCs (PMSCs)-treated group and a myelosuppression group.METHODS: The lungs from male C57BL/6 mice were digested with collagenase Ⅱ,followed by centrifugation over a Ficoll step.The interface fraction was collected and cultured by adherent method.When the monolayer of adherent cells reached 70%-80%confluence,the adherent cells were detected and expanded in culture medium.The mice in the PMSCs-treated and myelosuppression groups were intraperitoneally administered busulfan to inhibit the immigration of bone marrow stem cells.In addition,pulmonary fibrosis injury was induced in these mice with bleomycin.The PMSCs-treated group was intravenously administered 5×105 PMSCs.At the same time,the myelosuppression group received 100 μ L of phosphate buffered saline.After 14 days,the lungs were taken to prepare paraffin-embedded section.MAIN OUTCOME MEASURES: Cell morphology,immunophenotype,specific markers of the induced cells,expression of gene peroxisome proliferator activated receptor γ,osteopontin,osteocalcin,prosurfactant protein C(SP-C),Oct-4 and Nanog,histological alteration of lung tissue sections.RESULTS: MSCs were successfully isolated from mouse lung tissue.The pulmonary MSCs(PMSCs) were fibroblast-like cells,and expanded rapidly in vitro for up to 40 passages.The phenotype of the PMSCs was Scu- 1+ ,CD44+ ,CD29+ ,CD105+ ,CD54+ ,CD34-,CD45-,CD11b,c-kif,and CD31.They did not express alveolar epithelial cell specific markers,such as SP-C,aquaporin-5,and clara cell secretory protein.It was noticeable that stem cell markers octamer-binding transcription factor 4 (Oct-4) and Nanog were expressed continuously in culture expanded cells.They could differentiate into adipocytes,osteoblasts and alveolar epithelial cells in vitro.Colony-forming assay demonstrated that the colony forming efficiency of the PMSCs was nearly 3%.The cells from single colony were capable of differentiating to osteocytes,adipocytes and alveolar epithelial cells.Intravenous administration of PMSCs could alleviate pulmonary injury and fibrosis induced by bleomycin,despite the bone marrow was intact or not.CONCLUSION: The PMSCs are able to extensively propagate in vitro,can efficiently switch to both mesenchymal and alveolar epithelial lineages in vitro,generate adipocytes,osteocytes,and the alveolar epithelial cells,and reduce pulmonary injury and fibrosis in bleomycin challenged lung in vivo,strongly suggesting their promising applications in cellular therapy against the lung injury.

Objective: This study was aimed to explore antitumor responses induced by recombinant vaccinia viruses expressing a point mutant p53 (rVV-p53FL) and enhancive effects of recombinant vaccinia viruses expressing costimulatory molecule B7 (rVV-B7). Methods: A 135 Cys to Tyr point mutant p53 protein was used as the model of tumor associated antigen. rVV-of3FL and rVV-B7 were used as vaccines to test their induction of CTLs and antitumor immunity. Results: Immunization BABL/c mice with rVV-p53FL could elicited specific CD8+ CTLs that could effecively lyse P815-mp53 cells, a transfectant of the murine P815 mastocytoma containing the mutant p53 gene. Treatment with rVV-p53FL could survive a part of mice challenged with 1 ? 106 P815-mp53. Treatment with rVV-p53FL could significantly prolong survival of tumor-bearing force. Admixture at 1: 1 ratio of rVV-p53FL and rVV-B7 could enhance therapeutic antitumor effects of rVV-p53FL. ~Conclusion: Mutant P53 over-expressed in tumor cells can render cells targets for specific CTLs generated by immunization with mutant p53 protein based vaccine. Costimulatory molecule H7 can enhance tumor-associated antigen inducing antitumor responses.

Objective To explore the correlation of baseline CCL19+dendritic cell(CCL19+DC)infiltration in lung adenocarcinoma microenvironment with immunotherapy efficacy and CD8+T cell infiltration.Methods We retrospectively analyzed the data of patients with lung adenocarcinoma hospitalized at First Affiliated Hospital of Henan University of Science and Technology from January,2020 to December,2023,and collected tissue samples from 96 patients undergoing immunotherapy for assessing CCL19+DC and CD8+T cell infiltration using immunofluorescence assay.We evaluated the predictive value of baseline CCL19+DCs for patient responses to immunotherapy using receiver-operating characteristics(ROC)curves and analyzed the correlations of baseline CCL19+DC expression with immunotherapy efficacy and CD8+T cell and cytotoxic T lymphocyte(CTL)infiltrations.In co-culture systems of lung adenocarcinoma PC9 cells,CD8+T cells and DCs(overexpressing CCL19 with or without anti PD-1 antibody treatment),the expressions of granzyme B,perforin,IFN-γ,and Ki-67 in T cells were analyzed using flow cytometry.Results The patients with partial or complete remission following immunotherapy had a significantly higher baseline CCL19+DC infiltration level in lung adenocarcinoma tissues than those with poor responses.CCL19+DC infiltration had an area under ROC curve of 0.785,a sensitivity of 75.6%,and a specificity of 62.8%for predicting immunotherapy efficacy.The expression of CD8+T cell surface molecules Granzyme B(P<0.01),Perforin(P<0.01),IFN-γ(P<0.01)and Ki-67(P<0.001)in patients with high expression of CCL19+DC were higher than those in patients with low expression of CCL19+DC.The baseline CCL19+DC infiltration level was positively correlated with immunotherapy efficacy(P=0.003),CTL infiltration of(r=0.6657,P<0.001)and CD8+T cell infiltration(P=0.007).In the co-cultured cells,CCL19 overexpression combined with anti-PD1 treatment of the DCs more strongly enhanced cytotoxicity and proliferation of CD8+T lymphocytes than either of the single treatments(P<0.01 or 0.001).Conclusion The baseline CCL19+DC infiltration level in lung adenocarcinoma microenvironment is positively correlated with immunotherapy efficacy and CTL infiltration and can thus predict the response to immunotherapy.

Objective To explore the correlation of baseline CCL19+dendritic cell(CCL19+DC)infiltration in lung adenocarcinoma microenvironment with immunotherapy efficacy and CD8+T cell infiltration.Methods We retrospectively analyzed the data of patients with lung adenocarcinoma hospitalized at First Affiliated Hospital of Henan University of Science and Technology from January,2020 to December,2023,and collected tissue samples from 96 patients undergoing immunotherapy for assessing CCL19+DC and CD8+T cell infiltration using immunofluorescence assay.We evaluated the predictive value of baseline CCL19+DCs for patient responses to immunotherapy using receiver-operating characteristics(ROC)curves and analyzed the correlations of baseline CCL19+DC expression with immunotherapy efficacy and CD8+T cell and cytotoxic T lymphocyte(CTL)infiltrations.In co-culture systems of lung adenocarcinoma PC9 cells,CD8+T cells and DCs(overexpressing CCL19 with or without anti PD-1 antibody treatment),the expressions of granzyme B,perforin,IFN-γ,and Ki-67 in T cells were analyzed using flow cytometry.Results The patients with partial or complete remission following immunotherapy had a significantly higher baseline CCL19+DC infiltration level in lung adenocarcinoma tissues than those with poor responses.CCL19+DC infiltration had an area under ROC curve of 0.785,a sensitivity of 75.6%,and a specificity of 62.8%for predicting immunotherapy efficacy.The expression of CD8+T cell surface molecules Granzyme B(P<0.01),Perforin(P<0.01),IFN-γ(P<0.01)and Ki-67(P<0.001)in patients with high expression of CCL19+DC were higher than those in patients with low expression of CCL19+DC.The baseline CCL19+DC infiltration level was positively correlated with immunotherapy efficacy(P=0.003),CTL infiltration of(r=0.6657,P<0.001)and CD8+T cell infiltration(P=0.007).In the co-cultured cells,CCL19 overexpression combined with anti-PD1 treatment of the DCs more strongly enhanced cytotoxicity and proliferation of CD8+T lymphocytes than either of the single treatments(P<0.01 or 0.001).Conclusion The baseline CCL19+DC infiltration level in lung adenocarcinoma microenvironment is positively correlated with immunotherapy efficacy and CTL infiltration and can thus predict the response to immunotherapy.

Objective The objective of this study was to investigate the proliferation,autophagy and the potential mechanism of Ubiquitin-like with PHD and ring finger domains 1(UHRF1)in lung adenocarcinoma cells. Methods The expression of UHRF1 in lung adenocarcinoma tissues was determined by the bioinformatics website(TCGA). The expression of UHRF1 in lung adenocarci-noma cell lines(PC-9,A549 and H1299)and human bronchial epithelial cells(16HBE)was detected by qRT-PCR and Western blot. After transfection of UHRF1-shRNA,CCK-8,clone formation and ki67 were performed to detect the changes in the prolifera-tive capacity of lung adenocarcinoma A549 cells. Western blot was used to detect the changes of autophagy-associated proteins(LC3-I/II and Beclin-1)and proliferation-related proteins(CDK6,Rb and PCNA). Transmission electron microscopy was used to ob-serve the effect of UHRF1 on autophagosomes in A549 cells. Results The expression of UHRF1 in lung adenocarcinoma tissues was significantly higher than that in adjacent tissues. Compared with normal bronchial epithelial 16HBE cells,the mRNA and protein levels of UHRF1 in lung adenocarcinoma A549 and H1299 cells were significantly increased. In addition,CCK-8 assay and colony forma-tion experiments showed that silencing UHRF1 reduced the growth of A549 cells. Ki-67 immunofluorescence staining showed that the proliferation ability of A549 cells after knocking out UHRF1 was significantly lower than that in the normal control group. Further-more,knockdown of UHRF1 resulted in an increased expression of CKD6 and PCNA proteins in comparison with the control-siRNA group. The expression of Rb protein was down-regulated in the UHRF1-siRNA group. Silencing UHRF1 increased the ratio of LC3-II/LC3-I, induced up -regulation of Beclin -1 expression and promoted the formation of autophagic bodies in A549 cells. Conclusion UHRF1 is highly expressed in lung adenocarcinoma,and silencing UHRF1 can inhibit proliferation. This effect may be regulated by promoting autophagy.

Objective: To establish aNBR1-knockout lung cancer mouse model through CRISPR-Cas9 technology by using adeno-associated virus(AAV) as a vector to specifically inhibitNBR1expression and to investigate the impact ofNBR1knockout on tumor growth and immune cell infiltration and regulation. Methods: sgRNAs targeting mouseNBR1(Gene ID: 17966) was designed using the online tool CRISPOR(http://crispor.tefor.net/crispor.py). AAV6 was utilized as the vector for sgRNA delivery, and the efficiency of gene knockout was confirmed using PCR and DNA sequencing methods. To determine the best AAV infection approach in mice, 6 C57BL/6J mice were randomly divided into intranasal and endotracheal groups. After 28 days, lung tissue sections were assessed for enhanced green fluorescent protein expression to identify the more efficient infection method for subsequent experiments. Lung tumor growth, as well as immune cell infiltration and activation status in tumor tissues, were detected using methods including HE staining, immunohistochemistry, immunofluorescence, and flow cytometry. Results: DNA sequencing and immunofluorescence results indicated successful construction of the AAV6-U6-sgNBR1-CAG-Cre-GFP vector with stable knockout efficiency. Fluorescence microscopy showed higher efficiency of lung infection in mice through intratracheal administration(P<0.05). HE staining revealed reduced tumor area in mouse lungs after targetedNBR1knockout compared to the control group(P<0.01). Immunofluorescence and flow cytometry results demonstrated enhanced functional activity of CD8+T lymphocytes in lung cancer tissues of mice with targetedNBR1knockout, characterized by increased effector T lymphocytes and decreased exhausted T lymphocytes(P<0.01). Conclusion Using CRISPR/Cas9 technology, we construct a lung cancer mouse model with targetedNBR1knockout. We verify that targeted inhibition of NBR1 expression significantly enhances the functional activity of CD8+T lymphocytes in lung tissues, resulting in suppressed tumor growth, reduced tumor burden, and extended survival in lung cancer mice. This study lays an experimental foundation for investigations into the mechanisms and functions ofNBR1and other genes in lung adenocarcinoma cells.

Objective To investigate the mechanism through which RgpB,a virulence factor of Porphyromonas gingivalis(Pg),induces chemoresistance in esophageal squamous carcinoma.Methods The autophagy-regulating factors that interact with RgpB were screened by immunoprecipitation-mass spectrometry.The interaction between RgpB and the autophagy regulator TBC1D5 was investigated using co-immunoprecipitation.The impact of Pg infection on the expression of esophageal cancer cell membrane receptor molecule Cx43 was assessed using Western blotting.Immunofluorescence assay was used to analyze the relationship among Lamp1,Cx43 and TBC1D5.The effect of Pg infection on autophagosome-lysosome fusion was evaluated using autophagy double fluorescence technique.The effects of Pg infection and a Cx43 inhibitor on proliferation of esophageal cancer cells after chemotherapy were examined with plate cloning assay and CCK-8 method.Results Immunoprecipitation-mass spectrometry identified TBC1D5 as an autophagy regulator interacting with RgpB,and co-immunoprecipitation suggested that RgpB could directly bind to TBC1D5.In Pg-infected esophageal cancer cells,the expression of Cx43 on the cell membrane was significantly higher than that in non-infected cells.Immunofluorescence assay showed that the expression of Cx43 on the membrane of esophageal cancer cells increased significantly after Pg infection,which blocked autophagosome-lysosome fusion as shown by stubRFP-sensGFP-LC3 lentivirus study.Plate cloning assay and CCK-8 assay showed that the Cx43 inhibitor significantly attenuated the effect of Pg infection for promoting proliferation of esophageal cancer cells after chemotherapy.Conclusion Pg infection in esophageal cancer blocked autophagosome-lysosome fusion in the tumor cells,thereby preventing Cx43 from lysosomal degradation and leading to chemoresistance of esophageal cancer.

Objective To investigate the mechanism through which RgpB,a virulence factor of Porphyromonas gingivalis(Pg),induces chemoresistance in esophageal squamous carcinoma.Methods The autophagy-regulating factors that interact with RgpB were screened by immunoprecipitation-mass spectrometry.The interaction between RgpB and the autophagy regulator TBC1D5 was investigated using co-immunoprecipitation.The impact of Pg infection on the expression of esophageal cancer cell membrane receptor molecule Cx43 was assessed using Western blotting.Immunofluorescence assay was used to analyze the relationship among Lamp1,Cx43 and TBC1D5.The effect of Pg infection on autophagosome-lysosome fusion was evaluated using autophagy double fluorescence technique.The effects of Pg infection and a Cx43 inhibitor on proliferation of esophageal cancer cells after chemotherapy were examined with plate cloning assay and CCK-8 method.Results Immunoprecipitation-mass spectrometry identified TBC1D5 as an autophagy regulator interacting with RgpB,and co-immunoprecipitation suggested that RgpB could directly bind to TBC1D5.In Pg-infected esophageal cancer cells,the expression of Cx43 on the cell membrane was significantly higher than that in non-infected cells.Immunofluorescence assay showed that the expression of Cx43 on the membrane of esophageal cancer cells increased significantly after Pg infection,which blocked autophagosome-lysosome fusion as shown by stubRFP-sensGFP-LC3 lentivirus study.Plate cloning assay and CCK-8 assay showed that the Cx43 inhibitor significantly attenuated the effect of Pg infection for promoting proliferation of esophageal cancer cells after chemotherapy.Conclusion Pg infection in esophageal cancer blocked autophagosome-lysosome fusion in the tumor cells,thereby preventing Cx43 from lysosomal degradation and leading to chemoresistance of esophageal cancer.