Low Tumor Infiltrating Mast Cell Density Reveals Prognostic Benefit in Cervical Carcinoma.

Objectives: Research on the role of mast cells (MCs) in cervical tumor immunity is more limited. Therefore, our study aimed to evaluate the prognostic value of MCs and their correlation with the immune microenvironment of cervical carcinoma (CC). Methods: The Cancer Genome Atlas (TCGA) data was utilized to obtain the degree of immune infiltration of MCs in CC. Meanwhile, this study retrospectively collected patient clinical characteristic data and tissue specimens to further verify the relevant conclusions. Mast cell density (MCD) was measured by the CIBERSORT algorithm in TCGA data and immunohistochemical staining of tryptase in CC tissues. Finally, differentially expressed genes (DEGs) of TCGA data were performed using "limma" packages and key gene modules were identified using the MCODE application in Cytoscape. Results: The results showed MCs were diffusely distributed in CC tissues. Moreover, we found that low tumor-infiltrating MCD was beneficial for overall survival (OS) in the TCGA cohort. Consistent conclusions were also obtained in a clinical cohort. In addition, a total of 305 DEGs were analyzed between the high tumor-infiltrating MCD and low tumor-infiltrating MCD group. Seven key modules, a total of 34 genes, were screened through the MCODE plug-in, which was mainly related to inflammatory response and immune response and closely correlated with cytokines including CSF2, CCL20, IL1A, IL1B, and CXCL8. Conclusion: In short, high tumor-infiltration MCs in CC tissue was associated with worse OS in patients. Furthermore, MCs were closely related to cytokines in the tumor microenvironment, suggesting that they collectively played a role in the immune response of the tumor. Therefore, MCD may be a potential prognostic indicator and immunotherapy target of CC.

Comprehensive Analysis of the Expression and Prognosis for MCMs in Human Gastric Cancer.

PURPOSES: Minichromosome maintenance (MCM) proteins play an important role in replication and cell cycle progression. Even so, their expression and prognostic roles in cancer remain controversial. METHODS: To address this issue, the study investigated the roles of MCMs in the prognosis of GC by using ONCOMINE, GEPIA2, UALCAN, Cancer Cell Line Encyclopedia (CCLE), the Human Protein Atlas, Kaplan-Meier Plotter, cBioPortal, GeneMANIA, and DAVID databases. RESULTS: Over expressions of mRNA and cell lines were found in all members of the MCM family, and MCMs were found to be significantly associated with pathological tumor grades in GC patients. Besides, higher mRNA expressions of MCM1/5/7 were found to be significantly associated with shorter overall survival (OS) and progression-free survival (FP) in GC patients, while higher mRNA expression of MCM4/6/9 were connected with favorable OS and FP. Moreover, a high mutation rate of MCMs (68%) was also observed in GC patients. CONCLUSIONS: The results indicated that MCM1/5/7 were potential targets of precision therapy for patients with GC. And MCM4/6/9 were new biomarkers for the prognosis of GC. The results of the study will contribute to supplement the existing knowledge, and help to explore therapeutic targets and enhance the accuracy of prognosis for patients with GC.

Tumor-Associated CD163+ M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer.

BACKGROUND: Programmed death-ligand 1 (PD-L1) is a negative costimulatory molecule, and its main function is widely considered to be in the regulation of T cells. Tumor-associated macrophages (TAMs) are an important part of the tumor microenvironment, and they also play an important role in immunosuppression. However, the relationship between the expression of PD-L1 and TAMs in cervical carcinoma (CC) remains unclear. We detected the expression of PD-L1 and TAMs in tumor tissue to study the correlation between them. METHODS: Immunohistochemical staining of PD-L1, CD68 (pan-macrophage), and CD163 (M2-like macrophage) was performed in 120 cases of cervical squamous cell carcinoma. Logistic regression analysis was used to evaluate the predictors related to positive PD-L1 expression. We also apply the Kaplan-Meier method to study the recurrence-free and overall survival rate of CC patients. RESULTS: The increase in PD-L1 expression in tumor cells (TC) was significantly correlated with the increase in CD163 density (r=0.8550, p<0.0001), while PD-L1 in the stroma was also significantly associated with the intratumoral density of CD68- or CD163-positive cells (CD68 p<0.0001; CD163 p=0.0009). The mean infiltration rates of CD68- and CD163-positive cells in PD-L1-positive TC were significantly higher than in PD-L1-negative TC (CD68 p=0.0095; CD163 p<0.0001). In multivariate logistic regression analyses, only the density of CD163-positive cells was correlated with the expression of PD-L1 in TC cells (OR 1.52; p=0.032). In prognostic analysis, PD-L1 more than 10% was significantly correlated with short RFS (HR=2.66; p=0.028). For CD163+ macrophage evaluation, the density above the median was also significantly correlated with RFS (HR=2.48; p=0.021). CONCLUSION: CD163+ M2-like macrophage infiltration is highly associated with PD-L1 expression in CC, suggesting that macrophage infiltration can serve as a potential therapeutic target.

Effect of different types of macrophages on hepatic fibrosis in Echinococcus Granulosus mice.

OBJECTIVE: The purpose of this study is to illustrate the therapeutic effect of which kind of polarized macrophages-based cell therapy in hepatic fibrosis caused by cystic echinococcosis. METHODS: The isolation culture and polarization induction of mouse bone marrow-derived macrophages (BMDM) are established in an in vitro environment. A model of Echinococcus granulosus infection is established by direct injection of the Echinococcus granulosus suspension into the left hepatic lobe. The macrophages are labeled in vitro and the localization of the returned macrophages in the liver of the mice is determined by in vivo tracing. Macrophages of different polarization types are injected into the successfully modeled mice through the tail vein, and the results of HE, Masson, Sirius Red, Desmin immunohistochemistry and Hyp content are inspected to evaluate by liver tissue. Liver pathology and changes in the degree of fibrosis. RESULTS: Bone marrow-derived macrophages have been successfully obtained and induced into M1 and M2 macrophages by different conditions; a model of Echinococcus granulosus infection was successfully established. Macrophages labeled in vitro were returned to the model through the tail vein and they can be located in the liver; a variety of experimental results show that compared with the PBS group, the degree of fibrosis in the M0 group and the M1 group have been reduced, with statistical difference, and the M1 is better than M0 in terms of the therapeutic effect. There is no significant change in the degree of fibrosis in the M2 group. CONCLUSION: Both M1 and M0 macrophages can alleviate liver fibrosis caused by persistent infection of Echinococcus granulosus, but the treatment effect of M1 macrophages is more significant. Cell therapy based on M1 macrophages may be a new idea for treating liver fibrosis caused by persistent infection of Echinococcus granulosus.

The Leukocyte VCS Parameters Compared with Procalcitonin, Interleukin-6, and Soluble Hemoglobin Scavenger Receptor sCD163 for Prediction of Sepsis in Patients with Cirrhosis.

BACKGROUND: Patients with liver cirrhosis have a high risk of sepsis and a poor prognosis. Recently, a new standard for sepsis (Sepsis-3) has been proposed in the general population. The Coulter Lh 750 hematology analyzer can evaluate mean volume, conductivity, scatter, and distribution width of leukocyte. We tried to use Sepsis-3 criteria to study the diagnostic value of volume, conductivity, and scattering (VCS) parameters in sepsis and infection in patients with liver cirrhosis compared with traditional infection markers (PCT, IL-6, sCDl63). METHODS: A blinded, cohort study was conducted in three different ED populations within three affiliated hospitals. A total of 249 patients with liver cirrhosis were enrolled in the study. According to the "Sepsis-3" consensus criteria, clinical history, and laboratory examination, the subjects were divided into sepsis (n = 54), patients with infections (n = 95), and patients without systemic infections (n = 100). The blood samples of the patients were collected at the time of ED admission and were evaluated for the detection of sepsis. RESULTS: The differences of MNV, MNS, MMV, MMS, MLV, NDW, and MDW in the three groups were statistically significant. In the diagnosis of sepsis in patients with liver cirrhosis, the sensitivity of combined detection of MMV and MDW was 88.89%; the specificity was 74%. This sensitivity was significantly better than the 83.3% achieved using 0.97 mg/L as the cutoff for sCD163. In the diagnosis of infection in cirrhosis, the sensitivity of combination of MNV and MMS was increased to 86.32%; the specificity was 92%. The sensitivity was the same as that achieved by using 0.31 ng/mL as the cutoff value of PCT, but the specificity increased. CONCLUSION: The leukocyte VCS parameter could be potential parameters for indicating sepsis and infection in patients with liver cirrhosis. The combined detection of MMV and MDW seemed to be helpful for the diagnosis of sepsis in these patients, and the combination of MNV and MMS could better indicate infection for them.

The relationship between the PD-1/PD-L1 pathway and DNA mismatch repair in cervical cancer and its clinical significance.

BACKGROUND: According to recent clinical observations, deficient DNA mismatch repair (dMMR) is capable of improving antitumor effects of the PD-1/PD-L1 pathway, suggesting that dMMR may act as a prognostic indicator of PD-1/PD-L1 antibody drugs. In this study, we examined the dMMR and PD-1/PD-L1 expression, as well as explored the correlation of dMMR status with PD-1/PD-L1 expression in cervical cancer patients, in order to optimize cervical cancer patient selection for PD-1/PD-L1 antibody drug treatment, which is helpful to avoid adverse effects and keep costs manageable. METHODS: Sixty-six tissue samples from patients with squamous cell carcinoma were collected, and data of their clinical characteristics were also gathered. Based on these samples, the expression levels of MLH1, MSH2, and PD-L1 in cancer cells were tested by immunohistochemical assay (IHC). Moreover, PD-1/PD-L1 expression in tumor-invading lymphocytes (TILs) was detected by IHC as well. Six single-nucleotide-repeat markers of microsatellite instability (MSI), including NR-27, MONO-27, BAT-25, NR-24, NR-21, and BAT-26, were tested by capillary electrophoresis sequencer analysis. According to expression of MLH1, MSH2 and the MSI test, all 66 cases were divided into dMMR or proficient DNA mismatch repair (pMMR) groups. The comparisons of dMMR and PD-L1 in cancer cells and of PD-1/PD-L1 in TILs were conducted categorized by age, childbearing history, history of abortion, ethnicity, and cancer cell differentiation subgroup. Furthermore, PD-L1 levels in cancer cells and PD-1/PD-L1 in TILs were analyzed and compared in both dMMR and pMMR subgroups. RESULTS: Of the patient samples, 25.8% were associated with dMMR. PD-L1 in cancer cells, PD-L1 in TILs, and PD-1 in TILs took up 59.1%, 47.0%, and 60.6%, respectively. The data indicated that both dMMR and PD-L1 overexpression resulted from lower cancer differentiation, more incidences of childbearing, and a history of abortion. Abortion could significantly increase PD-1 expression levels in TILs. Additionally, more incidence of childbearing or older age (35-55 years) was able to upregulate PD-L1 expression in TILs. Statistical difference of PD-L1 in cancer cells could be observed between dMMR and pMMR subgroups. In the dMMR group, PD-L1 in cancer cells and PD-1 in TILs had no correlation (rs=0.161, p=0.537), but in the pMMR group, they had good correlation (rs=0.645, p<0.001). CONCLUSION: According to prior studies and our own experiments, PD-L1 in both cancer cells and TILs and PD-1 in TILs are widely observed in cervical cancer patients, indicating that there may be potential to apply PD-1/PD-L1 antibody drugs in cervical cancer. dMMR patients are associated with higher PD-L1 expression compared with pMMR ones, which suggested that PD-1/PD-L1 antibody drugs may work well in dMMR cervical cancer patients. Moreover, in patients with more incidences of childbearing or abortion, dMMR may be a molecular detection target for clinical application of PD-1/PD-L1 antibody drugs.

Transmembrane protein 166 expression in esophageal squamous cell carcinoma in Xinjiang, China.

OBJECTIVE: Transmembrane protein 166 (TMEM166) expression in esophageal squamous cell carcinoma (ESCC) and remote normal esophageal tissues was examined to assess any role in tumour biology. METHODS: TMEM166 mRNA expression in 36 cases with ESCC (36 tumour samples, 36 remote normal esophageal tissue samples) was detected by RT-PCR. TMEM166 protein expression was analysed in paraffin-embedded tissue samples from the same cases by immunohistochemistry. RESULTS: Semi-quantitative analysis showed TMEM166 mRNA expression in ESCCs to be significantly lower than in remote normal esophageal tissues (0.759 ± 0.713 vs. 2.622 ± 1.690, P=0.014). TMEM166 protein expression was also significantly reduced (69.4% vs. 94.4%, P<0.01). CONCLUSION: TMEM166 mRNA and protein expression demonstrated significant reduction in ESCCs compared with remote esophageal tissues, albeit with no correlation with tumour size, differentiation, stage, and lymph node metastasis, suggesting a role in regulating autophagic and apoptotic processes in the ESCC.

[Dynamic change of IL-10 and TGF-beta1 in the liver of Echinococcus multilocularis-infected mice].

OBJECTIVE: To observe the dynamic expression and function of IL-10 and TGF-beta1 in liver of BALB/c mice infected with Echinococcus multilocularis (Em). METHODS: Sixty female BALB/c mice were randomly divided into experiment group and control group. Mice in the experiment group were each injected with 0.2 ml Em protoscolex suspension (containing about 400 protoscoleces) , while those in control group received same volume of normal saline. At 2, 8, 30, 90, 180, and 360 d after infection, 5 mice from each group were sacrificed and liver specimens were collected for pathological examination and immunohistochemical detection for IL-10 and TGF-beta1. RESULTS: In mice of the experiment group, Em cysts in different sizes were found in the abdominal cavity and the liver tissue, which gradually enlarged with the time. HE staining showed infiltration of lymphocytes in liver tissue, pathological change between the cyst wall and hepatic cells. In the control, the liver lobules showed integrity and inflammatory cells were seen occasionally. The level of IL-10 expression in liver tissue of the infected mice increased with the time, and reached a peak [(1639 +/- 1.73) %] at 90 d post-infection and maintained a high level thereafter. The expression of TGF-beta1 also reached the highest level [(23.69 +/- 2.29) %] . Both were significantly higher than the control (P < 0.01), though a low level expression was found in the control at 90d post-injection. CONCLUSION: The expressions of IL-10 and TGF-beta1 both increase in the middle and late stages of the infection. Besides, their inhibited functions do not be helpful for clearing and controlling Echinococcus multilocularis infection in livers.

[Regulation on phenotypic and functional maturation of dendritic cells by apolipoprotein A-I].

AIM: to investigate the effects of apolipoprotein A-I (ApoA-I) on the peripherial blood dendritic cell (PBDC) and monocyte derived DC (MDDC) in vitro. METHODS: isolate PBDC or monocyte by cell isolation kit, monocyte were induced to MDDC by treated with GM-CSF plus IL-4 for 6 days, and then collect PBDC and MDDC treated them with apoA-I, LPS or TNF-α for 24 hours. Then check the cell surface marker and phagocytic capacity by flow cytometry. ELISA was used to detect the levels of cytokine secretion. T cells were stained with CFSE and T cell proliferation was assessed by flow cytometry. RESULTS: collect the PBDC and MDDC with high purity. In the presence of ApoA-I, the surface markers on MDDC, such as CD40, CD86 and MHC-II, were up-regulated which were detected by flow cytometry. CD83 expression on both PBDC and MDDC was remarkably increased. ApoA-I DC demonstrated decreased the phagocytic capacity. ApoA-I also stimulated MDDC to produce IL-12 and TNF-α. Furthermore, ApoA-I can induce considerable Th cell proliferation. CONCLUSION: ApoA-I can induce the maturation and activation of MDDC and PBDC, including the cytokine secretion, specific antigen presentation and T cell proliferation and decreasing the phagocytic capacity. Therefore, ApoA-I may attribute to the immune response in AS process.