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Hypoxia-inducible factor （HIF） are a family of molecules that are critical for regulating cellular adaptation to hypoxia, which can be activated by hypoxia and other stimuli, and then translocated into nucleus, where they modulate the transcription of target genes. Under the pathological conditions, such as inflammation and tumor, tissue microenvironment is often hypoxic. Innate lymphoid cells （ILCs） and T lymphocytes serve as important effector cells in defense against infection and tumor, and their functions are regulated by HIF proteins. In recent years, increasing evidence highlights the importance of HIF1α and HIF2α, two members of the HIF family, in regulating ILCs and T cells, providing new insights into mechanisms and treatment of diseases. This review summarizes the research progress on the role of HIF family proteins in ILCs and T cells and the underlying mechanisms. [ABSTRACT FROM AUTHOR]

Adoptive cell immunotherapy has been a hot spot in tumor research in recent years. Chimeric antigen receptor T cells (CAR-T) have achieved great success in hematological tumors and have changed the current tumor treatment landscape to a certain extent. However, the application of CAR-T therapy in clinics is limited due to its serious side effects and high treatment costs. Natural killer (NK) cells are important immune cells in the body and have native cytotoxicity and well safety. NK cells based on CAR engineering (CAR-NK) have shown powerful anti-tumor activity and safety in preclinical research and could be the next generation of CAR platform-based cellular immunotherapy. This review will systematically introduce the current research status of CAR-NK cells in lymphoma.

In this study, eight potential functional strains were selected to interfered with RAW264.7 murine macrophages and human peripheral blood mononuclear cells (PBMCs). Then, changes in phagocytic activity of RAW264.7 cells and natural killer (NK) cell activity were detected and the screened potential probiotics were further intervened in BALB/c mice to explore their immunomodulatory efficacy in vivo. In cell experiments, the results showed that the intervention of different strains significantly increased the phagocytic activity of RAW264.7 cells (P0.05). In conclusion, Bifidobacterium bifidum FL-228.1 can improve innate immune function and have a more comprehensive effect on the immune system by regulating immune cell activity, cytokine expression and mRNA levels of immune molecules related to antimicrobial peptides.

Adoptive cell immunotherapy has been a hot spot in tumor research in recent years. Chimeric antigen receptor T cells (CAR-T) have achieved great success in hematological tumors and have changed the current tumor treatment landscape to a certain extent. However, the application of CAR-T therapy in clinics is limited due to its serious side effects and high treatment costs. Natural killer (NK) cells are important immune cells in the body and have native cytotoxicity and well safety. NK cells based on CAR engineering (CAR-NK) have shown powerful anti-tumor activity and safety in preclinical research and could be the next generation of CAR platform-based cellular immunotherapy. This review will systematically introduce the current research status of CAR-NK cells in lymphoma. [ABSTRACT FROM AUTHOR]

Copyright of Science & Technology of Food Industry is the property of Science & Technology of Food Industry Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

We aimed to evaluate the immunological characteristics of different TCM patterns of primary immune thrombocytopenia( ITP) patients with failed first-line treatment and the response to eltrombopag. Methods From January, 2018 to September, 2021, the clinical data of 58 first-line failed ITP patients treated with eltrombopag were enrolled at First Affiliated Hospital of Zhejiang Chinese Medical University. Enrolled patients were divided into four groups based on TCM patterns, and their immune indicators, including lymphocyte subsets, immunoglobulin levels, and complement factors, with the efficacy of eltrombopag compared. Patients were classified into pattern of qi failing to control blood (22 cases), pattern of internal blockade of static blood(9 cases), pattern of bleeding due to blood heat (13 cases), and pattern of endogenous heat due to yin deficiency ( 14 cases) group based on TCM patterns. Results Comparative analysis of immune indexes of ITP patients with different TCM patterns showed that the percentage and absolute count of NK cells in the pattern of internal blockade of static blood group(16. 91%±7. 24%,(230. 33±188. 35) ×10 6 L -1 ) were significantly higher than those in the pattern of qi failing to control blood group(9. 76%±6. 33%,(109. 38± 76. 60)×10 6 L -1 ) and the pattern of bleeding due to blood heat group(5. 82%±3. 70%,(52. 50±40. 91) ×10 6 L -1 )(P<0. 05). However, there was no statistical difference in the levels of immunoglobulin and complement factors among all the groups. Comparison of the efficacy response of patients with different TCM patterns to eltrombopag showed that the effective rates in the pattern of qi failing to control blood, pattern of internal blockade of static blood, pattern of bleeding due to blood heat, and pattern of endogenous heat due to yin deficiency group were 68. 2%(15 / 22), 22. 2%(2 / 9), 61. 5%(8 / 13), and 78. 6%(11 / 14), respectively; the curative effect in the pattern of internal blockade of static blood group was different from that in the pattern of endogenous heat due to yin deficiency and pattern of qi failing to control blood group ( P < 0. 05). Conclusion The proportion of NK cells in patients’ peripheral blood may be involved in the difference of therapeutic response to eltrombopag in ITP patients with different TCM patterns. High NK cell counts are more common in ITP patients of the pattern of internal blockade of static blood group, which is related to the lower expected therapeutic response to eltrombopag. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To probe into the effects of Huiyang Guben decoction on natural killer ( NK) cell reconstruction after allogeneic hematopoietic stem cell transplantation ( allo-HSCT) and its molecular mechanism. METHODS: (1) Clinical experiment. Totally 40 patients undergoing allo-HSCT in the hospital from Jul. 2021 to Sept. 2022 were divided into the control group (n = 20) and HYGB group (n = 20) according to the treatment method. The HYGB group was intervened with Huiyang Guben decoction for 4 weeks before allo-HSCT. The efficacy of two groups was evaluated after allo-HSCT treatment. The peripheral blood NK cells were counted, the expression of peripheral blood NK cell marker molecules CD56 and CD16 was detected. The peripheral blood NK cells of two groups were isolated, the cell viability was measured in vitro, and the expression of phosphatidylinositol 3 kinase / protein kinase B (PI3K/ Akt) pathway protein was detected. (2) In vitro cell experiment. NK cells were isolated and cultured from the peripheral blood of healthy people, and then divided into the control group ( negative control ), HYGB group (intervention with Huiyang Guben decoction) and LY294002 group ( intervention with Huiyang Guben decoction + PI3K inhibitor LY294002). The cell viability and apoptosis of three groups were measured. RESULTS: (1) The good rate of hematopoietic reconstruction in the control group was 90%(18 / 20), the recurrence rate was 10%(2 / 20), and the nontransplantation-related mortality was 5%(1 / 20). The good rate of hematopoietic reconstruction in the HYGB group was 100%(20 / 20), the recurrence rate was 5% (1 / 20), and there was no death. There was no significant difference in efficacy between two groups ( P > 0. 05). However, the incidence of acute graft-versus-host disease (aGVHD) in the HYGB group was 10% ( 2 / 20), significantly lower than 45% ( 9 / 20) in the control group, the proportion of NK cells in peripheral blood and the expression of CD56 and CD16 were significantly higher than those in the control group, the proliferation activity of NK cells in vitro was higher than that in the control group, and the apoptosis rate was lower than that in the control group, with statistically significant differences (P<0. 05). It was also found that PI3K/ Akt pathway was significantly activated in NK cells in HYGB group compared with the control group (P<0. 05). (2) Compared with the HYGB group, the proliferation activity of NK cells in LY294002 group decreased and the apoptosis rate increased, with statistically significant differences (P<0. 05). LY294002 reversed the effect of Huiyang Guben decoction on promoting NK cell activity in vitro. CONCLUSIONS: Huiyang Guben decoction can promote the reconstruction and activity maintenance of NK cells after allo-HSCT by activating PI3K/ Akt pathway. [ABSTRACT FROM AUTHOR]

Copyright of Evaluation & Analysis of Drug-Use in Hospitals of China is the property of Evaluation & Analysis of Drug-Use in Hospitals of China Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

At present, the mechanism of chronic hepatitis B virus (HBV) infection has not been fully clarified, and HBV can cause immune cell dysfunction, immune imbalance and functional defects in the host, which might be one of the mechanisms of chronic HBV infection. In chronic HBV infection, the functions of natural killer (NK) cells are depleted, which may be associated with the upregulated expression of immune checkpoint molecules on the surface of NK cells, and blocking immune checkpoint molecules can restore the function of NK cells. This article systematically reviews the etiology of HBV, the mechanism of chronic HBV infection, and the effect of common immune checkpoint molecules on functions of NK cells in chronic hepatitis B, so as to provide new ideas for the treatment of chronic HBV infection. [ABSTRACT FROM AUTHOR]

Multiple myeloma is one type of hematological malignancy, characterized by the proliferation and accumulation of monoclonal plasma cells in bone marrow. Bone marrow microenvironment plays a key supporting role in the proliferation and survival of myeloma cells, where a large number of immune cells exist but are functionally suppressed. Based on the studies of myeloma immune microenvironment in recent years, we summarize the recent advances and existing problems in the treatment of multiple myeloma and put forward some considerations and suggestions, to provide references for researchers in this field.

Objective: To study the distribution characteristics of natural killer (NK) cells and their subsets in normal peripheral blood in China, and to explore their normal value and significance., Methods: In this study, peripheral blood was collected from 200 healthy adults. Their age range was 18-87 years. All the subjects were divided into 6 age groups: 18-30, 31-40, 41-50, 51-60, 61-70, and 71-87 years. With CD16, CD56, CD4, CD19, as surface markers, fluid cytology detection techniques were used to detect NK cells and the relative and absolute counts. SPSS 27.0 was used for systematic analysis of the data, and the measurement data were expressed as mean±standard deviations. A t test, variance analysis or rank sum test were performed to compare the differences between the age groups and the sex groups. The significance level was set at α=0.05, and P < 0.05 was considered statistically significant., Results: The range of NK B cells in the 200 healthy adult subjects was (0.46±0.24)×10 6 /L, that of CD3 - CD56 + NK cells was (13.14±7.56)×10 6 /L, that of CD56 dim CD16 + NK cells was (5.23±3.12)×10 6 /L, that of CD56 bright NK cells was (85.61±7.40)×10 6 /L, and that of NK T cells was (4.16±3.34)×10 6 /L. There were no statistically significant differences in CD3 - CD56 + NK cells and NK T cells with respect to age ( P = 0.417, P =0.217). However, there was a decreasing trend in the number of NK B cells and CD56 dim CD16 + NK cells with increasing age ( r =0.234, P < 0.001; r =0.099, P < 0.001), particularly after the age of 50. Conversely, CD56 bright NK cells showed an increasing trend with age ( r =0.143, P < 0.001)., Conclusion: The detection of NK cells and their subsets has significant reference value for the diagnosis, treatment, and prognosis of autoimmune diseases, infectious diseases, and tumors. This study provides a preliminary reference range for clinical detection of NK cell subsets, but further research with a larger sample size and multi-center trials are needed to confirm these findings.

Multiple myeloma is one type of hematological malignancy, characterized by the proliferation and accumulation of monoclonal plasma cells in bone marrow. Bone marrow microenvironment plays a key supporting role in the proliferation and survival of myeloma cells, where a large number of immune cells exist but are functionally suppressed. Based on the studies of myeloma immune microenvironment in recent years, we summarize the recent advances and existing problems in the treatment of multiple myeloma and put forward some considerations and suggestions, to provide references for researchers in this field. [ABSTRACT FROM AUTHOR]

Copyright of Practical Oncology Journal is the property of Journal of Practical Oncology Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective To investigate and screen key genes and related signaling pathways in the process of natural killer (NK) cells inhibiting the development and progression of hepatocellular carcinoma (HCC), Methods The gene microarray data GSE120123 was downloaded from the National Center for Biotechnology Information (NCBI) data-base Gene Expression Omnibus (GEO), and we selected 4 samples of NK cells, 2 of which were from normal liver tissues and 2 of which were from HCC tissues. The differentially expressed genes (DEGs) were analyzed and were input into the DAVID database for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis, and we constructed protein-protein interaction using Cytoscape software. Module analysis was also performed using STRING and Cytoscape. Finally, the key genes were selected by CytoHubba plug-in. Results A total of 317 DEGs (218 down-regulated genes and 99 up-regulated genes) were identified. GO and KEGG analysis showed that, leukocyte cell adhesion, negative regulation of RNA polymerase II promoter transcription, cytosolic adhesion molecules, Rap1 signaling pathway, cytokine-cytokine receptor interaction, chemokine signaling and other signaling pathways were found to be involved in the inhibition of development and progression of HCC by NK cells. The PPI network consisted of 268 nodes and 242 edges. Finally, 4 key genes were screened out, including CCR1, CCR5, CXCR6, and CXCL12. Conclusion CCR1, CCR5, CXCR6, and CXCL12 were key potential key genes of NK cells in inhibiting development and progression of HCC. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of ARL67156, a small-molecule inhibitor of CD39, on cytotoxicity of natural killer (NK) cells against gastric cancer cells., Methods: Human peripheral blood-derived primary NK cells isolated and purified using a magnetic bead antibody method were treated with 100 μmol/L ARL67156 for 24 h, and the signaling pathway of NK cell activation was detected by Western blotting. The level of interferon-γ (IFN-γ) in the supernatant of NK cells co-cultured with gastric cancer cells was detected using ELISA, and NK cell CD107a degranulation was measured with flow cytometry. The cytotoxicity of NK cells against co-cultured gastric cancer cells was evaluated using flow cytometry. In a nude mouse model bearing subcutaneous gastric cancer xenografts, the therapeutic effect of intravenous transfusion of NK cells and intraperitoneal injection of ARL67156 was assessed by measuring the changes in tumor volume., Results: (25.97 ± 5.69) % of peripheral blood NK cells from healthy individuals positive for CD39 expression. Treatment with ARL67156 significantly upregulated the activation molecules including NKG2D, DAP10, CD57, and CD16 and reduced the expressions of the inhibitory receptors TIGIT and KIR, thereby promoting the secretion of IFN-γ and CD107a degranulation in NK cells ( P < 0.05). In both the in vitro and in vivo experiments, ARL67156 significantly enhanced the cytotoxicity of NK cells against gastric cancer cells ( P < 0.05)., Conclusion: ARL67156 activates NK cells through the vav1-Syk signaling pathway to enhance their cytotoxicity against gastric cancer cells, which may serve as a new strategy for NK cell immunotherapy for gastric cancer.

Objective: To investigate the regulatory effect of interferon-α (IFN-α) on the apoptosis and killing function of CD56 dim CD57 + natural killer (NK) cells in systemic lupus erythematosus (SLE) patients, and to explore the specific mechanism., Methods: A total of sixty-four newly treated SLE patients and sixteen healthy controls (HC) enrolled in the Second Hospital of Dalian Medical University were selected as the research subjects. And the gene expression levels of molecules related to NK cell-killing function were detected by real-time quantitative polymerase chain reaction. CD56 dim CD57 + NK cells were co-cultured with the K562 cells, and the apoptotic K562 cells were labeled with Annexin-Ⅴ and 7-amino-actinomycin D. Peripheral blood mononuclear cells were treated with 20, 40, and 80 μmol/L hydrogen peroxide (H 2 O 2 ), and treated without H 2 O 2 as control, the expression level of perforin (PRF) was detected by flow cytometry. The concentration of IFN-α in serum was determined by enzyme linked immunosorbent assay. The expression levels of IFN-α receptors (IFNAR) on the surface of CD56 dim CD57 + NK cells were detected by flow cytometry, and were represented by mean fluorescence intensity (MFI). CD56 dim CD57 + NK cells were treated with 1 000 U/mL IFN-α for 24, 48 and 72 h, and no IFN-α treatment was used as the control, the apoptosis and the expression levels of mitochondrial reactive oxygen species (mtROS) were measured by flow cytometry and represented by MFI., Results: Compared with HC( n =3), the expression levels of PRF1 gene in peripheral blood NK cells of the SLE patients ( n =3) were decreased (1.24±0.41 vs . 0.57±0.12, P =0.05). Compared with HC( n =5), the ability of peripheral blood CD56 dim CD57 + NK cells in the SLE patients ( n =5) to kill K562 cells was significantly decreased (58.61%±10.60% vs . 36.74%±6.27%, P < 0.01). Compared with the control ( n =5, 97.51%±1.67%), different concentrations of H 2 O 2 treatment significantly down-regulated the PRF expression levels of CD56 dim CD57 + NK cells in a dose-dependent manner, the 20 μmol/L H 2 O 2 PRF was 83.23%±8.48% ( n =5, P < 0.05), the 40 μmol/L H 2 O 2 PRF was 79.53%±8.56% ( n =5, P < 0.01), the 80 μmol/L H 2 O 2 PRF was 76.67%±7.16% ( n =5, P < 0.01). Compared to HC ( n =16), the serum IFN-α levels were significantly increased in the SLE patients ( n =45) with moderate to high systemic lupus erythematosus disease activity index (SLEDAI≥10) [(55.07±50.36) ng/L vs . (328.2±276.3) ng/L, P < 0.001]. Meanwhile, compared with HC ( n =6), IFNAR1 expression in peripheral blood CD56 dim CD57 + NK cells of the SLE patients ( n =6) were increased (MFI: 292.7±91.9 vs . 483.2±160.3, P < 0.05), and compared with HC ( n =6), IFNAR2 expression in peripheral blood CD56 dim CD57 + NK cells of the SLE patients ( n =7) were increased (MFI: 643.5±113.7 vs . 919.0±246.9, P < 0.05). Compared with control ( n =6), the stimulation of IFN-α ( n =6) significantly promoted the apoptosis of CD56 dim CD57 + NK cells (20.48%±7.01% vs . 37.82%±5.84%, P < 0.05). In addition, compared with the control ( n =4, MFI: 1 049±174.5), stimulation of CD56 dim CD57 + NK cells with IFN-α at different times significantly promoted the production of mtROS in a time-dependent manner, 48 h MFI was 3 437±1 472 ( n =4, P < 0.05), 72 h MFI was 6 495±1 089 ( n =4, P < 0.000 1), but there was no significant difference at 24 h of stimulation., Conclusion: High serum IFN-α level in SLE patients may induce apoptosis by promoting mtROS production and inhibit perforin expression, which can down-regulate CD56 dim CD57 + NK killing function.

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objectives: Paired box gene 6 ( PAX6 ) plays a major role in the regulation of embryonic development. Abnormal expression of PAX6 is associated with the development of various tumors. PAX6 can play a role in promoting or suppressing cancer in different tumors. This study aim to observe the effect of overexpression of PAX6 on the growth of hepatocellular carcinoma cells, and the killing of hepatocellular carcinoma cells via natural killer (NK) cell and the possible mechanism., Methods: The protein levels of PAX6, soluble major histocompatibility complex class I-like protein A (sMICA) and soluble UL16 binding protein 2 (sULBP2) in peripheral blood from 68 cases of hepatocellular carcinoma (HCC) patients and 10 healthy volunteers were detected by ELISA. Hepatocellular carcinoma cell line (HepG2, LM3) and human normal liver cells (LO2) were cultured at 37 ℃ and 5% CO 2 condition in vitro. The PAX6 overexpressed plasmid (PAX6-OE) and empty vector (NC) were transferred into HepG2 and LM3 cells to construct stable cell lines. The mRNA and protein expression levels of PAX6 in HepG2 and LM3 cells were detected by real-time PCR, Western blotting and immunofluorescence, respectively. PAX6 was overexpressed in HepG2 and LM3 cells, the cell growth and migration ability were detected by CCK-8 method and cell scratch assay, and the levels of sMICA and sULBP2 in the supernatant were detected by ELISA. Matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9) and disintegrin and metalloproteinase 10 (ADAM10) in HepG2 and LM3 cells were detected by Western blotting. The killing ability of NK cells against these 2 HCC cells was detected by flow cytometry., Results: Compared with the healthy volunteers, the expressions of PAX6 in the HCC patients were significantly decreased ( P =0.002), while the expression of sMICA and sULBP2 were significantly increased ( P =0.004 and P <0.001, respectively). Real-time PCR and Western blotting results showed that compared with LO2 cells, mRNA and protein expressions of PAX6 in HepG2 and LM3 cells were significantly decreased (all P <0.05). Immunofluorescence results also showed that the expressions of PAX6 in HepG2 and LM3 were lower than those of LO2 cells. Compared with the NC group, the ability of proliferation and migration of HepG2 and LM3 cells were decreased (both P <0.05). The protein expressions of MMP2, MMP9 and ADAM10 in HepG2 and LM3 cells in the PAX6-OE group were significantly decreased, and the levels of sMICA and sULBP2 in superneant of HepG2 and LM3 cells in the PAX6-OE group were significantly lower than those in the NC group (all P <0.05). Flow cytometry results showed that compared with the NC group, the proportion of NK cells killing HepG2 and LM3 cells in PAX6-OE group was significantly increased (both P <0.05)., Conclusions: The expression of PAX6 is decreased in serum of HCC patients and hepatocellular carcinoma cell lines. Overexpression of PAX6 can inhibit the growth of hepatocellular carcinoma cells, enhance the killing efficiency of NK cells against hepatoma cells. The mechanism is related to the inhibition of the expression of metalloproteinase via PAX6 and the decrease of the secretion levels of sMICA and sULBP2.

Objective To observe the effect of NKG2D/RAE-1 pathway on mediating natural killer (NK) cell and CD8+T cell (both cells were referred to as NK/T cells) killing hypophysoma cells and to investigate the immune escape mechanism of hypophysoma from the cytological level .Methods The hypophysoma cell line AtT 20 was selected .The im-munofluorescent labeling method was used to detect the expression level of NKG 2D ligand RAE-1, its position and distribu-tion in the hypophysoma cells .NK/T cells were isolated by NK/T cell sorting kit, and NK/T cells were placed in the upper chamber of the Transwell chamber, the target cells AtT20 were placed in the lower chamber of the Transwell chamber and then were randomly divided into the control group and experimental group .The control group was co-cultured under normal conditions for 0, 4, 8, and 24 h. In the experimental group, the anti-RAE1 antibody was added to the culture medium, and the cells were co-cultured for 0, 4, 8, and 24 h. MTT assay was used to detect the inhibitory rate of cell proliferation at each time point .Results RAE-1 was clearly expressed in AtT 20 cell membrane, and there was a weak expression in the cytoplasm .The positive expression rate of RAE-1 protein in AtT20 cells was 100%, among which the strong expression rate was 30%(15/50), the moderate expression rate was 60%, the weak expression rate was 10%, and there was no neg-ative expression. In the control group, at 4, 8 and 24 h after culture, NK/T cell proliferation inhibition rate increased as compared with that at 0 h (all P<0.05).In the experimental group, the proliferation inhibition rate gradually increased at 0, 4, 8 and 24 h (all P<0.05), and at 8 and 24 h, the proliferation inhibition rates in the experimental group were sig-nificantly higher than those of the control group (all P<0.05).Conclusions NK/T cells kill the hypophysoma cells through NKG2D/RAE-1 pathway, meanwhile, RAE-1 has a negative feedback effect on NK/T cells and consequently in-hibits the killing activity of immune cells against the tumor cells .NKG2D/RAE1 pathway can be used as immunotherapy target for hypophysoma. [ABSTRACT FROM AUTHOR]

Objective: To investigate the characteristic functional changes of the decidual natural killer (NK) cells and γδ T cells, two immunocytes in the decidua, at the maternal-fetal interface in in vitro fertilization-embryo transfer (IVF-ET) pregnancy., Methods: Decidual samples were collected from 12 women of natural pregnancy (NP) and 32 women of IVF-ET pregnancy, who were enrolled in the NP group and the IVF-ET group, respectively. Then part of the decidual samples were paraffin-embedded for HE staining and immunofluorescence staining, while the rest of the samples were digested and Percoll was used for isolating decidual immunocytes (DICs) by gradient centrifugation. Flow cytometry was used to determine the cell counts of decidual NK cells and γδ T cells and the expression levels of their surface activation markers, CD69 and NKG2D in the NP and the IVF-ET groups. In addition, the expression levels of IFN-γ, TNF-α, IL-17A, and IL-10, the intracellular cytokines, and granzyme B, perforin, and granulysin, the cytolytic granules, were measured. The characteristic changes in the relevant immunological indicators were compared and analyzed., Results: HE staining of the tissue specimens showed that the typical structure of decidua was observed, and that lymphocytes were enriched in the decidua. Immunofluorescence staining showed that the percentage of decidual NK (dNK) cells in nucleated cells of the IVF-ET group was significantly lower than that of the NP group ( P <0.05). Flow cytometry analysis of DICs showed that, compared with those of the NP group, the percentage of dNK cells of the IVF-ET group was decreased ( P <0.05) and the expression levels of IL-10 and perforin were significantly decreased in the IVF-ET group ( P <0.05). However, there was no significant difference in the decidual γδ T (dγδT) cell count between the two groups. The expression of IL-10, IL-17A, and perforin was downregulated in the IVF-ET group ( P <0.05). There was no significant difference in the expression of IFN-γ, TNF-α, granzyme B, and granulysin, the cellular function indicators ( P >0.05)., Conclusion: The dNK cell count and the secretion of some intracellular cytokines of dNK and dγδT cells of women of IVF-ET pregnancy decreased to some degree, which suggests that certain changes may have taken place in the immunological microenvironment at the maternal-fetal interface. The specific effect of these changes on pregnancy outcomes needs further investigation., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Sciences).)

Objective: To amplify the plasmid pIRES2-EGFP-Livin encoding functional human Livin gene to transfect natural killer cell (NK) and gastric carcinoma cell line SGC-7901. and test the expression in NK and gastric carcinoma cell line SGC-7901. Methods: The plasmid pIRES2-EGFP-Livin encoding functional human livin gene was amplified, the concentration and the purity of plasmid pIRES2-EGFP-Livin were identified. NK from peripheral blood of healthy people were acquired And the plasmid pIRES2-EGFP-Livin was transfected into NK and gastric carcinoma cell line SGC-7901 using HP transfection reagent in vitro. The transfection efficiency and the expression of human livin in NK and gastric carcinoma cell line SGC-7901 were compared. Results: A large quantity of NK cells were successfully amplified by serum-free medium in vitro; amounts of no endotoxin plasmid were acquired from Plasmid Midi Kit; plasmid had no mutations with high concentration and purity. The green bands of EGFP expression could be clearly observed in pIRES2-EGFP-Livin-transfected gastric carcinoma cell line SGC-7901. while could not be observed in NK. Conclusion: Effective expressions of Livin protein had been verified in gastric carcinoma cell line SGC-7901 transfected by Plasmid pIRES2-EGFP-Livin but not in NK. [ABSTRACT FROM AUTHOR]

Natural killer cells(NK) as the key components of the innate immune system are enriched in liver and have organ specificity on immunophenotype and function that show anti-infection and anti-tumor responses. Under normal circumstances, the stimulatory ligands expressed on the target cell interact with the activated receptors and stimulate NK cells to release cytotoxic substances and induce target cells apoptosis. However tumor cells are able to escape the immune surveillance of NK cells through a variety of ways.Over the past decade, growing evidence indicates that the abnormal expression of tumor-associated antigens, cytokines and interactions with other immune cells in the tumor microenvironment affected cytotoxic activity of natural killer cells, and played a central role in immune escape of tumor cells. This review focuses on recent studies about the natural killer cells involved in immune escape of liver tumor and may lead to provide reference points for antitumor therapeutic targets. [ABSTRACT FROM AUTHOR]

Objective: To observe the effect of Natural killer cells on hyperlipidemia of patients with gestational diabetes. Methods: A total of 95 gestational diabetes patients with hyperlipidemia, confirmed in Affiliated Hospital of Qiingdao University during February 2013 to April 2015, were chosen as research group; 100 healthy pregnant women from a medical physical examination, as control group at the same time. The levels of blood glucose, glycosylated hemoglobin, body mass index,total cholesterol, triglycerides, low density lipoprotein cholesterol, high density lipoprotein cholesterol, interleukin-2, IL-6, interferon-gamma, NK cells and the cell subtype were compared between the two groups. The correlation of NK cells with blood fat was analyzed by the Person correlation analysis. Results: (1) The levels of fasting blood glucose, 2 h postprandial blood glucose, glycosylated hemoglobin, BMI in the research group were signifi- cantly higher than those in the control group, the differences were statistically significant (P<0.05); (2) The levels of TC, TG, LDL -C in the research group were significantly higher than those in the control group, the level of HDL-C was significantly lower than that of the control group, the differences were statistically significant (P<0.05); (3) The levels of IL-2, IFN-γin the research group were significant- ly lower than those in the control group, with statistically significant differences (P<0.05); the level of IL-6 in the research group was sig- nificantly higher than that in the control group, with statistically significant differences (P<0.05); (4)The number of NK cells, CD4+/CD3+, CD4+/CD8+ CD56++CD4+/CD3+ in the research group were lower than those in the control group (P<0.05); CD8+/CD3+, CD18+/CD8+, CD19+/CD3+ were higher than those in the control group (P<0.05), there was no statistical significance in CD28+/CD8+ between the two groups (P>0.05). (5) NK cell count was negatively correlated with the levels of TC, TG, LDL-C (r=-0.527, -0.598, -0.485), positively correlated with HDL-C (r=0.528). Conclusion: The number of NK cells in the pregnant women with gestational diabetes declines, their activity also reduces, which might be correlated with change of different phenotypes of NK cells participating in the body's immune response, leading to blood lipid regulation imbalance and the process of hyperlipemia. [ABSTRACT FROM AUTHOR]

Objective: To investigate the changes of subpopulation and function of natural killer (NK) cells in adult patients with infectious mononucleosis related liver injury., Methods: Seventeen healthy controls, twenty-six patients of infectious mononucleosis (IM) without liver injury, and twenty-one IM patients with liver injury were enrolled in this study. Peripheral blood was collected. Plasma and peripheral blood mononuclear cells (PBMC) were purified. Plasma interleukin-15 (IL-15) level was measured by enzyme linked immunosorbent assay. Three NK cell subpopulations, including CD56 + CD16 - , CD56 + CD16 + , and CD56 - CD16 + NK cells, were assessed by flow cytometry. PBMC were co-cultured with 721.221, K562, and P815-Ab cells, respectively. CD107a expression and CD16 mean fluorescence intensity (MFI) in NK cells was investigated by flow cytometry. PBMC from IM patients with liver injury were stimulated with anti-IL-15 neutralizing antibody, and the change of NK cell function was observed. One-way ANOVA and paired t test were used for statistical analysis., Results: The percentage of CD56 + CD16 - NK cells in IM patients with liver injury (7.36±0.92)% and IM patients without liver injury (10.67±1.37)% were lower than that of controls (12.02±2.51)% ( P <0.05). CD56 + CD16 - NK cell frequency in IM patients with liver injury also was lower than that in IM patients with liver injury ( P <0.0001). The percentage of CD56 + CD16 + NK cells in IM patients with liver injury (62.03±8.78)% and IM patients without liver injury (44.74±13.03)% were higher than that in controls (40.74±10.15)% ( P <0.05). CD56 + CD16 + NK cell frequency in IM patients with liver injury also was higher than that in IM patients with liver injury ( P <0.0001). There was no statistical difference of CD56 - CD16 + NK cell proportion among three groups ( P =0.427). Plasma IL-15 level in IM patients with liver injury (309.1±68.00) pg/ml were higher than that in IM patients without liver injury (269.6±53.57) pg/ml and controls (257.4±73.93) pg/ml ( P <0.05). NK cells induced target cell death through different receptors upon activation, the ratio of CD107a-positive cells in NK cells of IM patients with liver injury was higher than that in IM patients without liver injury and controls ( P <0.05), while CD16 MFI in NK cells in IM patients with liver injury was lower than that in IM patients without liver injury and controls ( P <0.05). The percentage of CD107a positive cells in NK cells was significantly reduced in response to anti-IL-15 neutralizing antibody stimulation in IM patients with liver injury ( P <0.05)., Conclusion: IM patients with liver injury presented imbalance of NK cell subpopulation and enhanced NK cell function. The elevation of IL-15 might contribute to promote NK cell cytotoxicity in IM patients with liver injury.

Objective To investigate the effect of growth hormone on the cytotoxicity of natural killer (NK) cells from human peripheral blood mononuclear cells (PBMC) and the possible mechanism. Methods NK cells, purified by mag­netic activated cell sorting (MACS) from 10 volunteers, were divided into three groups. Cells in the groups A， B and C were co-cultured with recombinant human growth hormone ( rhGH) (A:0 μg/mL，B:5 μg/mL，C:20 μg/mL). After 48 h，the cytotoxicity of NK cells against K562 cells, with different effector-target ratio (25：1 , 50： 1，100:1 )，was analyzed by CCK-8. Real-time RT-PCR and flow cytometry were used to detect the expression of natural killer cell group 2 member D ( NKG2D)，in the levels of gene and protein molecules. Results The cytotoxicity of NK cells against K562 cells in the group B and C was significantly higher than that in the group A at different effector-target ratio，and group C was higher than group B (all P <0.01). The percentages of NK cells with positive expression of NKG2D in the groups A，B and C were 78. 8% ±2.8%，87. 5% ±3.1% and 91.8% ±3. 3%，respectively，and group B and C were higher than group A (all P <0.05 ). The relative expression of NKG2D mRNA of NK cells in the groups A，B and C was 1 ±0.01，5 ±2.32 and 10 ± 3. 21 respectively. The expression of NKG2D mRNA in the group B and C was significantly higher than that of group A，and group C was higher than group B ( all P <0.05 ) . Conclusion Growth hormone may enhance the cytotoxici­ty of NK cells from PBMC by up-regulating the expression of NKG2D. [ABSTRACT FROM AUTHOR]

Objective: To explore the significance of lymphocytes in systemic sclerosis (SSc), by detecting the levels of T lymphocytes, B lymphocytes and natural killer (NK) cells, and analyzing the correlation between the lymphocytes and clinical laboratory indexes., Methods: The numbers and proportion of T, CD4 + T, CD8 + T, B, and NK cells were detected by flow cytometry in peripheral blood of 32 SSc patients who had taken immunosuppressive drugs and 30 healthy controls (HC). The comparison of the lymphocyte subsets in SSc with them in the HC groups, and the correlation between the lymphocytes and other clinical and laboratory indicators were analyzed by the relevant statistical analysis., Results: Compared with the HC group, the numbers of T, CD4 + T, CD8 + T, and NK cells in peripheral blood of SSc group, who had taken immunosuppressive drugs, were significantly decreased ( P < 0.05). More-over, the proportion of NK cells in peripheral blood of the SSc group was also significantly lower than that in the HC group ( P =0.004). In addition, all the lymphocyte subsets were decreased in peripheral blood of more than 65% of the SSc patients who had taken immunosuppressive drugs. Compared with CD4 + T normal group, the positivity of Raynaud's phenomenon, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) was significantly increased in CD4 + T reduction group, respectively ( P ＝0.024, P < 0.001, P =0.018). ESR was higher in CD8 + T reduction group than CD8 + T normal group ( P ＝0.022). The prevalence of fingertip ulcer was significantly increased in B cell decrease group ( P =0.019). Compared with NK cell normal group, the prevalence of fingertip ulcer was significantly increased in NK cell lower group ( P =0.033), IgM was remarkablely decreased yet ( P =0.049). The correlation analysis showed that ESR was negatively correlated with the counts of T lymphocytes ( r ＝－0.455, P ＝0.009), CD4 + T lymphocytes ( r ＝－0.416, P ＝0.018), CD8 + T lymphocytes ( r ＝－0.430, P =0.014), B cells ( r ＝－0.366, P ＝0.039)., Conclusion: The number of CD4 + T, CD8 + T, B, and NK cells significantly decreased in peripheral blood of SSc patients who had used immunosuppressive drugs, some lymphocyte subsets might be related with Raynaud's phenomenon and fingertip ulcer, and reflected the disease activity by negatively correlated with ESR and CRP; the numbers of lymphocyte subsets in peripheral blood should be detected regularly in SSc patients who had taken immunosuppressive drugs.

Objective: To assess the effect of neutralizing CD96 on natural killer (NK) cell functions in mice with pulmonary Chlamydia muridarum infection and explore the possible mechanism., Methods: Male BALB/c mice were randomly divided into infection group (Cm group), anti-CD96 treatment group (anti-CD96 group) and control group ( n =5). In the former two groups, C. muridarum was inoculated via intranasal administration to establish mouse models of pulmonary C. muridarum infection, and the mice in the control group received intranasal administration of the inhalation buffer. In anti-CD96 group, the mice were injected with anti-CD96 antibody intraperitoneally at the dose of 250 μg every 3 days after the infection; the mice in Cm group received intraperitoneal injections of saline. The body weight of the mice was recorded daily. The mice were sacrificed 5 days after C. muridarum infection, and CD96 expression was detected by quantitative real-time PCR and Western blotting. HE staining and pathological scores were used to evaluate pneumonia of the mice. The inclusion body forming units (IFUs) were detected in the lung tissue homogenates to assess lung tissue chlamydia load. Flow cytometry and ELISA were used to assess the capacity of the lung NK cells to produce interferon-γ (IFN-γ) and regulate macrophages and Th1 cells., Results: C. muridarum infection inhibited CD96 expression in NK cells of the mice. Compared with those in Cm group, the mice in antiCD96 mice showed significantly milder lung inflammation ( P &lt; 0.05) and reduced chlamydia load in the lung tissue ( P &lt; 0.05). Neutralizing CD96 with anti-CD96 significantly enhanced IFN-γ secretion by the NK cells ( P &lt; 0.05) and augmented the immunoregulatory effect of the NK cells shown by enhanced responses of the lung macrophages ( P &lt; 0.05) and Th1 cells ( P &lt; 0.05)., Conclusions: Inhibition of CD96 alleviates pneumonia in C. muridarum -infected mice possibly by enhancing IFN-γ secretion by NK cells and augmenting the immunoregulatory effect of the NK cells on innate and adaptive immunity.

Objective: The Clinicopathological characteristics and the reason of misdiagnosis about nasal extranodal NK/T cell lymphoma in head and neck（ENKTL） are analysised to raise awareness and reduce misdiagnosis. Method: One hundred and twenty patients with ENKTL, diagnosed pathologically in head and neck from May 2010 to April 2018, were analyzed retrospectively. All cases were divided into misdiagnosed group and non-misdiagnosed group according to whether there were misdiagnosis and mistreatment before diagnosis. The differences of clinicopathological characteristics between the two groups before and after diagnosis were compared. The differences of overall survival（OS） between the two groups after treatment were also compared. Result: The misdiagnosis rate of this study was 71.7%, and they were misdiagnosed as sinusitis, nasal polyps, upper respiratory tract infection and so on. On the whole, the 1-year OS, 3-year OS and 5-year OS were 77.8%, 65.9% and 49.3% respectively. There was no significant difference in sex, age, first symptom, initial site, B symptom, superficial lymph node enlargement and weight loss between two groups before diagnosis（ P >0.05）, however, there were significant differences in relative specific signs, hospital grade and time from first visit to definite diagnosis（ P <0.05）. The proportion of the high-level hospitals in the first visit hospital was significantly lower than that in the non-misdiagnosed group, while the time from the first visit to the final diagnosis was significantly longer than that in the non-misdiagnosed group. There was no significant difference in international prognostic index（IPI） score, expression of Ki-67, therapeutic schedule, chemotherapy regimen for patients receiving chemotherapy, and radiation dose for patients receiving radiotherapy after diagnosis between the two groups（ P >0.05）, but there was a significant difference in the clinical stages of Ann Arbor between the two groups（ P <0.05）. The clinical staging of the misdiagnosed group was later than that of the non-misdiagnosed group. The 1-year OS, 3-year OS and 5-year OS were 73.6%, 59.3% and 43.2% in the misdiagnosed group respectively, and 88.2%, 82.4% and 64.2% in the non-misdiagnosed group respectively. The overall survival rate in the misdiagnosis group was lower than that in the non-misdiagnosed group, and the overall survival time was lower than that in the non-misdiagnosed group. However, there was no significant difference in the overall survival rates between the early stage（Ⅰ and Ⅱ） and the advanced stage（Ⅲ and Ⅳ） of Ann Arbor（ P >0.05）. Conclusion: The non-specific clinical and the complexity of pathological features of ENKTL in head and neck might cause misdiagnosis, which leads to prolonged time from first visit to definite diagnosis, thus results in disease progression, and ultimately may lead to decreased overall survival rate. To avoid the misdiagnosis and early diagnosis is very important., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: To investigate the effect of epithelial-mesenchymal transition (EMT) on biological activity of natural killer (NK) cells in esophageal squamous cell carcinoma (ESCC)., Methods: Western blot selected EMT EC9706 and non EMT KYSE150 from six esophageal cancer cell lines. NK cells were collected from 10 cases of healthy volunteers. According to different co-culture conditions, the medium was divided into 1640 normal medium stimulus-NK group (NC group), EC9706 cell supernatant stimulus-NK group (EC9706 group) and KYSE150 cell supernatant stimulus-NK group (KYSE150 group). The expression of NK cells surface/intracellular molecules was detected by flow cytometry after co-culture of NK cells with different conditioned medium for 72 h. 10 ng/mL transforming growth factor-β (TGF-β) treated KYSE150 cell line for 7 d to induce EMT. KYSE150 EMT and KYSE150 supernatant were collected and co-cultured with NK cells, respectively. 72 h later, flow cytometry was used to detect the expression of surface/intracellular molecules of NK cells, degranulation ability of CD107a, killing effect of target cell K562, proliferation and apoptosis of NK cells., Results: Two EMT-treated and four non-EMT-treated esophageal squamous cell carcinoma lines were selected from the six strains, and one EC9706 and one KYSE150 respectively were selected for subsequent experiments. The purity of NK cells was more than 90% and Tcells <1%. After the supernatant of esophageal squamous cell carcinoma cells was stimulated, the surface activation type and inhibitory type receptors of NK cells in the three groups were stimulated. The effector molecule results showed that: compared with NC group and KYSE150 group, the expressions of activated type receptors NKP30, NKG2D and NKP44 in EC9706 group were decreased, and the release of granulozyme was decreased ( P <0.05). Expressions of inhibitory receptor NKG2A and CD158b increased ( P <0.05). NKP46, CD226, CD16 expressions and perforin release showed no statistically significant difference among the three groups. Compared with KYSE150 supernatant stimulation, the expressions of activated receptors NKP30, NKG2D and NKP44 decreased after KYSE150 EMT supernatant stimulation, and perforin release decreased. The degranulation of CD107a and the killing effect of target cell K562 were decreased, and the proliferation index of NK cell Ki67 was decreased ( P <0.05). Expressions of inhibitory receptor NKG2A and CD158b increased ( P <0.05). The expressions of NKP46, CD226 and CD16, granulozyme release and apoptosis of NK cells were not statistically significant., Conclusion: EMT of esophageal cancer cells can escape the immune surveillance by inhibit the activity of NK cells and reduce the release of effective molecules., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)