Objective To explore the relationships between resistin rs1477341 , rs3219175 and type2 diabetes mellitus (T2DM) in Guangdong. Methods 180 patients of diagnosed T2DM in the Second Affiliated Hospital of Guangzhou Medical University from January 2013 to December 2014 were enrolled in the study as the treatment group and another 180 subjects taking health examination as the control group. The genotype distributions of rs1477341 locus A/T, rs3219175 locus A/G were compared and the relationship between the genotypes and T2DM were anlysed by the logistic regression. Results The frequencies of the rs1477341, rs3219175 locus genotype reached the genetic equilibrium, showed their representativeness of a population group. The frequency distribution of rs1477341 genotype was statistically insignificant across the two groups (P > 0.05). The frequency of rs3219175 genotype AA+AG was decreased while genotype GG increased significantly compared to the control group (P < 0.05). The logistic regression results suggest genotype GG may increase disease risk of T2DM which the morbidity rate of AG homozygote was 0.630 times as high as that of GG homozygote (OR =0.630, 95%CI:0.405 ~ 0.980, P < 0.05) and the morbidity rate of GG homozygote was 1.551 times as higher as that of AA+AG homozygote (OR = 1.551, 95%CI:1.016 ~ 2.368, P < 0.05). Conclusions rs1477341 genotype has no correlation with T2DM among the population in Guangdong , but rs3219175 genotype does. GG may have a higher susceptibility to T2DM.

Newcastle disease virus-like particles (NDV VLPs) are composed of matrix protein (M) as the skeleton,with the insertion of hemagglutinin-neuraminidase and/or fusion protein.NDV VLPs are reported to be immunogenic and can induce specific humoral and cellular immune responses.However,its relationship with innate immunity remains elusive.Dendritic cells (DCs) are a group of specialized antigen presenting cells,which are crucial in connecting innate immunity and adaptive immunity.In this study,NDV VLPs and murine DCs were used to investigate the connection between NDV VLPs and innate immunity.The DC maturation induced by NDV VLPs (M+ HN) was evaluated.The results showed that NDV VLPs could be effectively taken up by DC and presented to naive T cells.NDV VLPs-induced DC significantly up-regulated the expression of MHC Ⅱ and costimulatory molecules on DC surface,and subsequently promoted the secretion of proinflammatory cytokines.This experiments also showed that different assembled NDV VLPs induced significant stimulating ability in cytokine levels.In summary,NDV VLPs can induce DC maturation,which gives insights to better understanding of VLPs-mediated innate immunity and provide information in selecting preferred NDV VLPs candidate.

Objective:To explore the relationship between ABCB1 or ABCG2 gene polymorphisms and therapeutic effects of gefitinib in non-small cell lung cancer (NSCLC) patients, and establish a prediction model of efficacy.Methods:A total of 176 NSCLC patients with epidermal growth factor receptor (EGFR) -sensitive mutation treated with gefitinib admitted to Department of Pulmonary Disease of Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine of Hebei Province from December 2018 to December 2020 were employed as subjects, and all patients were detected ABCB1 and ABCG2 gene polymorphisms. Patients were divided into remission group and non-remission group according to curative effect after 3 months of gefitinib treatment. The clinical data, ABCB1 and ABCG2 gene polymorphisms, levels of serum carcinoembryonic antigen (CEA) and carbohydrate antigen 125 (CA125) were compared between the two groups. The related factors of failure to remission after treatment were analyzed by multivariate logistic regression analysis. Combined with ABCB1 and ABCG2 gene polymorphisms, the prediction model for gefitinib efficacy was constructed and the nomogram was drawn.Results:During the follow-up period, 5 patients were lost to follow-up and 7 patients withdrew from the trial due to intolerable adverse effects, finally 108 patients were employed as remission group, and 56 patients were employed as non-remission group. The numbers of GG, GT and TT at ABCB1 rs2032582 in the remission group were 49, 50 and 9, and those in the non-remission group were 12, 35 and 9, with a statistically significant difference ( χ2=9.56, P=0.008). The numbers of GG, GA and AA at ABCG2 rs2231137 in the remission group were 13, 72 and 23, and those in the non-remission group were 11, 42 and 3, with a statistically significant difference ( χ2=7.74, P=0.021). Before treatment, the levels of serum CEA in the remission group and the non-remission group were (34.28±5.11) ng/ml and (37.88±7.05) ng/ml, with a statistically significant difference ( t=3.74, P<0.001). The levels of CA125 of the two groups were (27.24±6.50) U/ml and (33.31±6.09) U/ml, with a statistically significant difference ( t=-5.79, P<0.001). Multivariate logistic regression analysis showed that TT at rs2032582 of ABCB1 gene ( OR=12.99, 95% CI: 3.17-53.23, P<0.001), GG at rs2231137 of ABCG2 gene ( OR=7.75, 95% CI: 1.36-44.07, P=0.021) and GA ( OR=6.94, 95% CI: 1.47-32.84, P=0.015), CA125 ( OR=1.18, 95% CI: 1.10-1.28, P<0.001) were independent risk factors of failure to remission in NSCLC patients with EGFR sensitive mutation after treatment. The consistency index (C-index) of nomogram for predicting failure to remission was 0.92 (95% CI: 0.86-0.94) . Conclusion:ABCB1 rs2032582 and ABCG2 rs2231137 polymorphisms are related to therapeutic effects of gefitinib in the NSCLC patients, the nomogram based on the two genes combined with serum CA125 can predict efficacy of gefitinib.

Objective To analyze the infiltration abundance of macrophage M2 in breast cancer tissues and explore the correlation between VSIG4 and macrophage M2 and the potential mechanism of regulating the invasion and migration of breast cancer patients. Methods We downloaded the RNA-seq data of TCGA-BRCA and assessed the infiltration abundance of immune cells in the samples by CIBERSORT, and established a prognostic risk prediction model. Then, we analyzed the effect of macrophage M2 and VSIG4 on the prognosis of breast cancer patients. In addition, we analyzed the signaling pathway associated with VSIG4 by gene set enrichment analysis and predicted its upstream regulation of miRNA. Results The infiltration abundance of macrophage M2, age, PR status and pathological stage were involved in the establishment of risk prediction model, and the model had a good prediction performance (AUC=0.816). High infiltration of macrophage M2 (HR=1.35, P < 0.05) and high expression of VSIG4 (HR=1.4, P=0.039) suggested poor prognosis of breast cancer patients. VSIG4 could be regulated by upstream miR-29a-3p and significantly correlated with Toll-like receptor, cell adhesion, production and release of cytokine. Conclusion VSIG4 is significantly associated with breast cancer patients' prognosis and infiltration of macrophage M2, regulated by the upstream miR-29a-3p and promotes the invasion and migration of breast cancer cells. It can be used as a potential prognostic marker for breast cancer.

OBJECTIVETo observe the cytotoxity of CD138-CAR-T cells on human multiple myeloma cell RPMI8226 and U266 cells and explore the impact of pomalidomide on the cytotoxity of CD138-CAR-T on RPMI8226 and U266 cells.

METHODSThe cytotoxity of CD138-CAR-T and CD138-CAR-T combined pomalidomide on RPMI8226 and U266 was detected by CFSE/7AAD. The effctor cells were co-cultured with target cells at 5:1 for 18 h, and then the supernatant were collected and used for ELISA assays.

RESULTSAfter 18 h co-culture, the cytotoxity of CD138-CAR-T on RPMI8226 and U266 was significantiy higher than control (P<0.01). There was no significant change on the cytotoxity of pomalidoide combined with CD138-CAR-T on RPMI8226 and U266. The results showed that co-cultured system contribted to a markedly increased production of IFN-γ, after adding pomalidomide to the co-cultured system. It can significantly enhance the production of IFN-γ, compared with CD138-CAR-T alone.

CONCLUSIONCD138-CAR-T had significantly cytotoxity on U266 and RPMI8226. Pomalidomide could promote CD138-CAR-T cells IFN-γ production.

The nucleocapsid protein (N protein) has been found to be an antigenic protein in a number of coronaviruses. Whether the N protein in severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is antigenic remains to be elucidated. Using Western blot and Enzyme-linked Immunosorbent Assay (ELISA), the recombinant N proteins and the synthesized peptides derived from the N protein were screened in sera from SARS patients. All patient sera in this study displayed strong positive immunoreactivities against the recombinant N proteins, whereas normal sera gave negative immunoresponses to these proteins, indicating that the N protein of SARS-CoV is an antigenic protein. Furthermore, the epitope sites in the N protein were determined by competition experiments, in which the recombinant proteins or the synthesized peptides competed against the SARS-CoV proteins to bind to the antibodies raised in SARS sera. One epitope site located at the C-terminus was confirmed as the most antigenic region in this protein. A detailed screening of peptide with ELISA demonstrated that the amino sequence from Codons 371 to 407 was the epitope site at the C-terminus of the N protein. Understanding of the epitope sites could be very significant for developing an effective diagnostic approach to SARS.
Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; chemistry ; immunology ; Humans ; Nucleocapsid Proteins ; chemistry ; immunology ; Peptide Fragments ; chemical synthesis ; Plasmids ; Recombinant Proteins ; immunology ; isolation & purification ; metabolism ; SARS Virus ; genetics ; immunology ; metabolism

Objective To observe and compare the effect of Cordyceps sinensis ( bailing ) capsules com-bined with weight-bearing breathing exercises, and weight-bearing breathing exercises combined with tiotropium bro-mide and seretide, on patients with stable but moderate-to-severe chronic obstructive pulmonary disease (COPD). Methods Sixty-three patients with moderate-to-severe COPD were randomly divided into an observation group and a control group. Both groups performed weight-bearing breathing exercises, supplemented in the observation group with the oral administration of bailing capsules. The control group instead inhaled tiotropium bromide and seretide. Six-mi-nute walking distance, the COPD assessment test ( CAT scores) and concentrations of interleukin-6 ( IL-6) , interleu-kin-8 ( IL-8) and tumor necrosis factor-α ( TNF-α) were observed after 1 ( T1) , 30 ( T2) and 58 ( T3) days of the treatments. Results At T1 there were no significant differences between the two groups in any of the measurements ( P≤0.05) . At T2, there were still no significant differences except that a significant decrease in IL-8 and TNF-αlevels was observed in the control group. At T3 the average CAT scores had decreased significantly in both groups compared to before the treatment but there was no significant difference between the two groups. In the observation group, the average 6MWT distance had increased significantly compared to before the treatment and compared to the control group, where there was no significant improvement. The average IL-6, IL-8 and TNF-αreadings of the control group were significantly lower than those of the observation group at T3 and compared to before the treatment. No sig-nificant changes in those indicators were observed in the observation group at T3. Conclusions Bailing capsules combined with weight-breathing exercises are more effective for relieving dyspnea symptoms and improving exercise capacity than weight-breathing exercises combined with tiotropium bromide and seretide. However, in controlling air-way inflammation and airway hyper-responsiveness, the triple inhalation combined with weight-bearing breathing exer-cises is more effective.

In comparison with the traditional two-dimensional tumor cell culture, the three-dimensional tumor spheroid culture can not only provide with an in vivo-like growth environment for tumor cells, but also maintain maximum cell activities. Therefore, the three-dimensional tumor spheroid culture is widely used in oncology research. In particular, the three-dimensional tumor cells retain the material and structural basis of the microenvironment of tumor in vivo, which is closer to the actual physiological environment, allowing it to be an ideal in vitro model for evaluating the tumor treatment response and drug resistance in tumors. This review summarizes the mechanisms of drug resistance in three-dimensional tumor cells, especially those induced by the morphology and microenvironment of three-dimensional tumor spheres, and puts forward the problems existing in the current three-dimensional tumor cells model, as well as the future development direction.

To build a CT-based radiomics predictive mode to evaluate the differentiation degree of the esophageal squamous carcinoma.
 Methods: A total of 160 patients with surgical pathology, complete clinical data and chest CT scanning before operation were retrospectively collected from January 2008 to August 2016. All patients were assigned randomly to a primary data set and an independent validation. Texture analysis was performed on CT images, while the carcinomas were performed by manual segmentation to extract the radiomics features. Radiomics features were extracted and 9 radiomics signatures were finally selected after dimension reduction. Radiomics features were extracted and established via Matlab. Multivariable logistic regression analysis was performed to build the predictive model. A 10-fold cross-validation was used for selecting parameters in the least absolute shrinkage and selection operator (LASSO) model by minimum criteria. The receiver operating characteristic (ROC) curves and areas under ROC curve (AUC) were used to compare the model performance in the primary validation and the independent validation for evaluating the differentiation degree of esophageal squamous carcinoma.
 Results: Radiomics signature showed great effect in discriminating primary data set and independent validation. The predictive model had a good performance in primary data set. The AUC was 0.791, the sensitivity was 81.6%, and specificity was 72.3%. In the independent validation, the AUC was 0.757, the sensitivity was 70.0%, and the specificity was 73.0%.
 Conclusion: The predictive model can be used for evaluating the differentiation degree of esophageal squamous carcinoma efficiently, which can be helpful to clinicians in diagnosis and choice of treatment for esophageal squamous carcinoma.
Carcinoma, Squamous Cell ; Esophageal Neoplasms ; Humans ; ROC Curve ; Retrospective Studies ; Tomography, X-Ray Computed

[Abstract] Objective： ： To explore the impact of γ-chain (γc) family cytokines (IL-2, IL-7, IL-15, IL-21) on T cell phenotypes in ex vivo culture to provide experimental evidence for ex vivo cell preparation in adoptive immunotherapy. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood of healthy volunteers; nylon column sorting, CD3+ magnetic beads sorting, CD3- magnetic beads sorting and natural sedimentation were used to sort T cells from PBMCs. The purity, recovery rate and viability of T cells sorted by the above methods were compared. The CD3/CD28 magnetic beads-activated CD3+T cells were cultured inAIMV medium with IL-2 or mixed cytokines (IL-7, IL-15, IL-21). The expansion fold and phenotypes of T cells in ex vivo culture were detected by flow cytometry. Results： ： The purity of T cells sorted by CD3- magnetic beads sorting was significantly higher than that sorted by nylon column, CD3+ magnetic beads sorting and natural sedimentation ([94.06±1.07]% vs [86.74±1.06]%, [89.61±1.40]%, [88.48 ± 1.86]%, P<0.05); The recovery rate of T cells sorted by natural sedimentation was significantly higher than that by other three methods ([60.29±1.53]% vs [45.03±2.79]%, [20.15±3.41]%, [42.98±2.82]%, P<0.05). Comprehensively, the natural sedimentation method is the best option. The ex vivo expansion fold of T cells in IL-2 group was significantly higher than that in mixed group ([262.6±143.2] times vs [73.0±25.8] times, P<0.05). The proportions of early memory T cells, Tscm+Tscm-like and Tcmin the mixed group were significantly higher than those in the IL-2 group ([55.6±1.82]% vs [39.6±1.52]%, [16.6±1.82]% vs [9.8±1.30]%, [39.0±1.58]% vs [29.2±1.79]%; all P < 0.05). Conclusion： ： Natural sedimentation sorting has advantages of low cost, high recovery and purity. Mixed cytokines of IL-7, IL-15 and IL-21 are beneficial for production of early memory T cells. This study provides an experimental data of ex vivo T cell preparation for cancer adoptive immunotherapy.