免疫细胞特征与细菌性肺炎风险遗传因果关系的孟德尔随机化研究

目的应用孟德尔随机化(Mendelian randomization,MR)技术探究免疫细胞特征与细菌性肺炎之间的相关性。方法选取已知731种免疫细胞特征,从全基因组关联研究(genome-wide association study,GWAS)开放数据库获取数据,使用双样本MR方法揭示免疫细胞特征是否对细菌性肺炎发生风险具有直接影响。两样本MR分析主要采用逆方差加权法(inverse variance weighting,IVW),根据效应指标优势比(OR)和95%置信区间(CI)评估结果,并进行敏感性分析,包括但不限于结果的稳健性验证、潜在的异质性检验以及多效性检验等确保最终结论的科学性及准确性。为避免反向因果关系,将细菌性肺炎作为暴露因素,筛选的六种免疫细胞特征作为结局事件,进行反向MR分析。结果通过MR分析识别出CD33^(-)HLA-DR^(+)髓样细胞、CD86^(+)树突状细胞、CD39^(+)调节性T细胞、HLA-DR^(+)自然杀伤细胞、CD14^(+)CD16^(+)单核细胞及程序性死亡配体1(PD-L1)单核细胞等六种检验效能P<0.01的免疫细胞特征与细菌性肺炎风险存在因果关联。反向MR结果未发现细菌性肺炎与上述六种免疫细胞特征存在因果关系。结论本研究揭示了遗传因素在免疫细胞特征及功能与细菌性肺炎发病风险之间的关系,筛选并鉴定了多种与细菌性肺炎存在显著联系的免疫细胞特征。

免疫细胞特征与骨质疏松症因果关系的双向孟德尔随机化研究

目的:探讨免疫细胞特征与骨质疏松症(osteoporosis,OP)的因果关系。方法:分别从IEU OpenGWAS project数据库和FINNGEN数据库中筛选并获得731个免疫细胞特征的全基因组关联研究(genome-wide association study,GWAS)数据集和OP的GWAS数据集。基于工具变量筛选标准,筛选符合要求的免疫细胞特征的单核苷酸多态性(single nucleotide polymorphism,SNP)位点和OP的SNP位点。将筛选的免疫细胞特征的SNP位点作为工具变量,采用逆方差加权法(inverse variance weighted,IVW)、MR-Egger、加权中位数、简单模式和加权模式等进行正向孟德尔随机化(Mendelian randomization,MR)分析,评估免疫细胞特征与OP的因果关系。采用MR-presso检验进行水平多效性检验,采用Cochran’s Q检验评估IVW法和MR Egger法分析结果的异质性,采用留一法评估MR分析结果的稳定性。将筛选的OP的SNP位点作为工具变量,以正向MR分析获得的与OP具有可靠因果关系的免疫细胞特征为结局进行反向MR分析。结果:32个免疫细胞特征与OP存在可靠的因果关系,其中包含7个绝对细胞计数(absolute cell counts,AC)特征、13个中位荧光强度(median fluorescence intensity,MFI)特征、12个相对细胞计数(relative cell counts,RC)特征。在7个AC特征中,Sw mem AC、IgD^(-)CD38dim AC、HLA DR^(+)NK AC与OP呈负向因果关系,CD62L-myeloid DC AC、CD33br HLA DR+AC、DN(CD4^(-)CD8^(-))AC、CD25^(++)CD8br AC与OP呈正向因果关系;在13个MFI特征中,BAFF-R on IgD^(-)CD38br、CD3 on CD8br、CD3 on CD39^(+)CD4^(+)、CD16-CD56 on NK、CD28 on CD4 Treg、CD16 on CD14^(-)CD16^(+)monocyte、CD8 on TD CD8br与OP呈负向因果关系,CD19 on IgD-CD38br、CD86 on myeloid DC、HLA DR on CD14^(+)CD16^(-)monocyte、HLA DR on CD14^(+)monocyte、CD45 on CD33br HLA DR^(+)CD14^(-)、HLA DR on CD33br HLA DR^(+)CD14dim与OP呈正向因果关系;在12个RC特征中,IgD^(+)CD38dim%lymphocyte、CD11c^(+)CD62L^(-)monocyte%monocyte、TD CD8br%CD8br、CD39^(+)CD8br%T cell与OP呈负向因果关系,IgD-CD38dim%B cell、CD62L-DC%DC、CD8br%leukocyte、CD8br and CD8dim%leukocyte、NKT%T cell、NKT%lymphocyte、HLA DR^(+)CD8br%lymphocyte、CD3-lymphocyte%leukocyte与OP呈正向因果关系。结论:部分免疫细胞特征与OP之间存在因果关系,这为探究免疫系统与OP间的作用机制提供了线索和方向。

Culture and Identification of Human Amniotic Mesenchymal Stem Cells

Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.

Prognostic significance of cell infiltrations of immunosurveillance in colorectal cancer

AIM: To determine whether the mast cell (MCs) and tumor-associated macrophage (TAMs) counts have any correlation with clinical outcome in colorectal cancer, and to investigate whether MCs undergo phenotypic changes in colorectal cancer.METHODS: The MC and TAM counts were determined immunohistochemically in 60 patients with colorectal cancer and the depth of invasion, lymph node metastasis rate, distant metastasis rates, and survival rates were compared between patients with low (less than the mean number of positive cells) and high (more than the mean number of positive cells) cell counts.RESULTS: Both patients with a low MC count and patients with a low TAM count had significantly deeper depth of invasion than those with a high MC count and those with a high TAM count (P＜0.01 and P＜0.01 respectively).Patients with a high MC count and patients with a high TAM count were significantly higher showing significantly lower rates of lymph node metastasis, distant metastasis than those with a low MC count and those with a low TAM count. There were significant positive correlation between MC counts and TAM counts (r = 0.852, P＜0.01).In both cancerous tissue and normal colorectal tissue,the predominant MC phenotype was MCTC. The 5-year survival rate estimated was significantly lower in both patients with a low MC count and patients with a low TAM count than in those with a high MC count and those with a high TAM count (P＜0.05 and P＜0.01 respectively).CONCLUSION: There appears to be a direct relationship between the number of MCs and clinical outcome in patients with colorectal cancer, even though MCs exhibited no significant phenotypic changes. TAM count is of value to predict the clinical outcome or prognosis. It is more beneficial for estimating biological character of colorectal carcinoma to combine MC and TAM counts.

The expression of Elf-1 and Ki-67 and their correlations in non-small-cell lung cancer

Objective: Elf-1 is a member of the proto-oncogenes Ets-related transcription factor family and over-expressed in many human tumors, Ki-67 is an important nuclear antigen associated with cell proliferation. This study investigated the expression of Elf-1 and Ki-67 in non-small-cell lung cancer(NSCLC) and studied their correlation with the clinicopathological features. Methods: Tissue microarray from 64 cases lung cancer tissue and 10 cases normal lung tissue was constructed, immunohistochemical method was used to evaluate the protein expressions of Elf-1 and Ki-67, correlations of the expression of Elf-1 and Ki-67 to clinicopathological features of NSCLC were analyzed. Results: Expression of Elf-1 and Ki-67 in NSCLC tissues were significantly higher than in normal lung tissues(P < 0.05), the positive rate of Elf-1 and Ki-67 was 73.44% and 64.06% in NSCLC group, Overexpression of Elf-1 in NSCLC was significantly related to histopathological grading, different clinical staging and the intensity of ELF-1 expression was significantly higher in the group with lymph node metastasis than that without(P < 0.05). Overexpression of Ki-67 was also closely related to tumor differentiation, clinical stages and lymph node metastasis(P < 0.05). In addition positive correlation was found between the expressive intensity of Elf-1 and Ki-67(τ = 0.295, P = 0.018). Conclusion: The high expression and positive correlation of Elf-1 and Ki-67 in NSCLC suggest that they probably play a role in onset and progression of lung cancer, united detecting their expression could be used as an valuable molecular biological index for predicting the malignant behavior and early diagnosis of NSCLC.