Hyperuricemia remission after colorectal cancer surgery for colorectal cancer patients.

The purpose of this study was to investigate whether patients with colorectal cancer (CRC) combined with hyperuricemia remitted 1 year after CRC surgery. CRC patients combined with hyperuricemia who underwent radical surgery were included from a single clinical center from Jan 2016 to Dec 2021. Baseline characteristics was compared between the remission group and the non-remission group. Multivariate logistic regression was used to find the possible predictive factors of hyperuricemia remission. A total of 91 patients were included for data analysis, retrospectively. There were 34 (37.4%) patients in the remission group and 57 (62.6%) patients in the non-remission group. The mean preoperative weight and body mass index (BMI) were 61.2 ± 10.7 (kg) and 24.1 ± 3.3 (kg/m2). 21 (23.1%) patients had a history of drinking. We found that the weight and BMI were not significantly different before and 1 year after CRC surgery (P > 0.05). In contrast, uric acid values were significantly decreased (P < 0.01). Meanwhile, the outcomes showed there were no significant differences in the baseline characteristics between the remission and non-remission groups (P > 0.05). According to multivariate logistic regression, we found that the history of drinking was a predictive factor of hyperuricemia remission (OR = 0.046, 95% CI 0.005-0.475, P = 0.010). CRC patients with hyperuricemia had a 37.4% remission from hyperuricemia 1 year after CRC surgery. Tumor location, tumor stage, and tumor size did not predict the remission of hyperuricemia. Notably, the history of drinking was a predictive factor of hyperuricemia remission.

A Novel Prognostic Chemokine-Related lncRNAs Signature Associated with Immune Landscape in Colon Adenocarcinoma.

Chemokines have been reported to be involved in tumorigenesis and progression and can also modulate the tumor microenvironment. However, it is still unclear whether chemokine-related long noncoding RNAs (lncRNAs) can affect the prognosis of colon adenocarcinoma (COAD). We summarized chemokine-related genes and downloaded RNA-seq and clinical data from The Cancer Genome Atlas (TCGA) database. A total of 52 prognostic chemokine-related lncRNAs were screened by univariate Cox regression analysis; patients were grouped according to cluster analysis results. Lasso regression analysis was applied to determine chemokine-related lncRNAs to construct a risk model for further research. This study first investigated the differences between the prognosis and immune status of two chemokine-related lncRNAs clusters by consensus clustering. Then, using various algorithms, we obtained ten chemokine-related lncRNAs to construct a new prognostic chemokine-related lncRNAs risk model. The risk model's predictive efficiency, validity, and accuracy were further validated and determined in the test and training cohorts. Furthermore, this risk model played a vital role in predicting immune cell infiltration, immune checkpoint gene expression, tumor mutational burden (TMB), immunotherapy score, and drug sensitivity in COAD patients. These findings elucidated the critical role of novel prognostic chemokine-related lncRNAs in prognosis, immune landscape, and drug therapy, thereby providing valuable insights for prognosis assessment and personalized treatment strategies for COAD patients.

Early-phase insulin hypersecretion associated with weight loss outcome after LSG: a prospective cohort study in Asian patients with BMI ≥28 kg/m2.

BACKGROUND: Obesity has become a global problem that poses a serious threat to human health. Laparoscopic sleeve gastrectomy (LSG) is an effective long-term treatment. However, the weight loss of some patients after LSG is still insufficient. It is necessary to investigate the factors associated with inadequate weight loss after LSG. OBJECTIVE: The objective of this study was to explore whether preoperative insulin secretion could be associated with weight loss after LSG in patients with obesity. SETTING: This is a single-center prospective cohort study conducted in a university hospital. METHODS: Patients from a prospective database who underwent LSG were analyzed. All 178 participants underwent a 75-g oral glucose tolerance test (OGTT) to assess preoperative insulin and c-peptide secretion before LSG. The areas under the curve (AUCs) for glucose, insulin, and c-peptide were determined in the OGTT. The percentage of excess weight loss (%EWL) and the percentage of total weight loss (%TWL) were used to estimate the effect of weight loss after LSG. Regression models were used to assess the correlation between preoperative insulin and c-peptide secretion with %EWL ≥75% and TWL ≥35% at 12 months after LSG. RESULTS: The AUCs of insulin and c-peptide were significantly lower in the %EWL ≥75% and %TWL ≥35% groups at 0-30 minutes, 0-60 minutes, and 0-120 minutes during the OGTT. At 30, 60, and 120 minutes during the OGTT, c-peptide levels were significantly lower in the %EWL ≥75% group and %TWL ≥35% group. The preoperative c-peptide level at 30 minutes during the OGTT (C30) was significantly negatively correlated with %EWL (ß = -.37, P < .001) and %TWL (ß = -.28, P = .011). Univariate logistic regression analysis showed that preoperative C30 was associated with %EWL ≥75% and %TWL ≥35% after LSG. According to multiple logistic regression analysis, patients with a low preoperative C30 had an 8-fold higher %TWL ≥35% after LSG than those with a high C30 (odds ratio: 8.41 [95% confidence interval: 1.46-48.58], P = .017). Similarly, patients with a low preoperative C30 had a 7-fold higher EWL% ≥75% after LSG than patients with a high C30 (odds ratio: 7.25 [95% confidence interval: 1.11-47.50], P = .039). CONCLUSIONS: The rate of weight loss after LSG is low among patients with preoperative hyperinsulinemia. The preoperative c-peptide level at 30 minutes during the OGTT is associated with weight loss after LSG.

R848/TLR7-Mediated Stronger CD8+ T Immunity Is Dependent on DC-NK Cell Interactions.

BACKGROUND: Toll-like receptor (TLR) 7 agonists are effective candidates for Th1 immune adjuvants, which compensate for the insufficient Th1 immune responses induced by traditional adjuvants. This effect is currently dependent on TLR7-mediated induction of dendritic cell (DC) maturation and increased IL-12 production. METHODS: In vivo, we intraperitoneally injected TLR agonists with OVA, and LNs were collected for detection. In vitro, Activated DCs, natural killer (NK) cells, and CD8+ T cells were tested using flow cytometry for surface expression and enzyme-linked immunosorbent assay for cytokine production. NK cell migration was evaluated using transwell system. All experiments were performed in both C57BL/6 and BALB/C backgrounds. RESULTS: Our findings revealed that the enhanced CD8+ T immunity characterized by CD8+ T accumulation, proliferation, and IFN-γ+CD8+ T induction induced by R848 was attributed to DC-dependent NK cell migration and DC-NK interactions. Our results demonstrated that R848 induced CD8+ T cell accumulation and IFN-γ+CD8+ T cells in lymph nodes (LNs) to a greater degree in vivo than TLR4 agonists (lipopolysaccharide) and TLR9 agonists (Class C CPG). R848-activated DCs enhanced CD8+ T cell proliferation and increased IFN-γ+CD8+ T cells with the assistance of NK cells. In contrast, depletion of NK cell decreased IFN-γ+CD8+ T cell production. Greater NK cell migration to LNs occurred in R848-immunized mice. A similar effect of R848 on NK cell migration was observed in an in vitro transwell study. When co-cultured, NK cells plus R848 could promote DCs maturation, and in turn, DCs in combination of R848 augmented NK cells activation. Further studies demonstrated that among several TLR agonists, R848 produced the largest amount of the chemokine CXCL9 from activated DCs, which is relevant to NK cell migration. CXCL9 blockade reduced the number of migrated NK cells, and the addition of CXCL9 increased the number of NK cells. DISCUSSION: Taken together, R848-mediated stronger CD8+ T cell immunity does not depend on DC activation alone, rather that NK cells must also be considered. By increasing our immunological understanding of the effect of R848/TLR7, these findings provide a new perspective for applying R848 in future clinical studies.

Transcutaneous Electrical Acupoint Stimulation Combined with Dexamethasone and Tropisetron Prevents Postoperative Nausea and Vomiting in Female Patients Undergoing Laparoscopic Sleeve Gastrectomy: a Prospective, Randomized Controlled Trial.

BACKGROUND: Despite the administration of prophylactic antiemetics, some patients who undergo laparoscopic sleeve gastrectomy (LSG) remain at high risk for postoperative nausea and vomiting (PONV). Although many trials have been conducted, the effectiveness of transcutaneous electrical acupoint stimulation (TEAS) on the prevention of PONV remains unknown. METHODS: Sixty-two female patients undergoing elective LSG were randomly assigned to the TEAS combined with dexamethasone and tropisetron (TEAS group, n = 31) or dexamethasone and tropisetron (control group, n = 31) groups. The incidence and severity of PONV, as well as the need for rescue antiemetics, were collected within 48 h after surgery. RESULTS: The patients in both groups had similar clinical characteristics and underwent the same surgical procedure. In the TEAS group, 13 patients (41.9%) had PONV within 48 h after LSG compared to 24 patients (77.4%) in the control group (P = 0.004, relative risk: 0.39 [0.19, 0.80]). The severity of PONV differed significantly between groups, with five patients (16.1%) in the TEAS group and 15 patients (48%) in the control group experiencing clinically important PONV (P = 0.007, relative risk: 0.62 [0.42, 0.90]). Moreover, fewer patients required antiemetic rescue medication in the TEAS group compared with the control group (29.0% vs. 58.1%, P = 0.021). CONCLUSION: Multimodal antiemetic prophylaxis consisting of TEAS and antiemetics was effective in reducing PONV incidence and intensity in high-risk patients undergoing LSG.

Neuropilin-1 Interacts with Fibronectin-1 to Promote Epithelial-Mesenchymal Transition Progress in Gastric Cancer.

INTRODUCTION: Neuropilin-1 (NRP1) binds to many ligands and co-receptors and affects cell survival and migration, which is essential for tumor progression. However, there are still largely unknowns about how NRP1 affects the epithelial-mesenchymal transition (EMT)-related malignant progression in gastric cancer. METHODS: We used TCGA to analyze the expression of NRP1 in gastric cancer and its impact on patient survival. In in vitro experiments, transwell, wound healing and colony formation assays were used to evaluate the effects of NRP1 and ginsenoside Rg3 on the invasion, migration and proliferation of gastric cancer cells. In in vivo experiments, we evaluated the overexpression and knockdown of NRP1 and the effect of ginsenoside Rg3 on tumor growth. RESULTS: We found that NRP1 is highly expressed in advanced gastric cancer and associated with poor prognosis. Knockdown of NRP1 expression can inhibit the proliferation and metastasis of gastric cancer cells. Mechanically. NRP1 interacts with fibronectin-1 (FN1) to promote the malignant progression of gastric cancer cells through ECM remodeling. In addition, we found that ginsenoside Rg3 can block the interaction of NRP1 and FN1 and inhibit the progression of gastric cancer. CONCLUSION: Our study suggested that the interaction of NRP1 and FN1 is crucial for the malignant progression of gastric cancer. This may provide a new perspective and potential treatment methods for the treatment of gastric cancer.

[Resiquimod (R848) has more stronger immune adjuvantivity than other tested TLR agonists].

Objective To compare and characterize the Th1 immune responses induced by the most commonly used commercial Toll-like receptor (TLR) agonists through in vivo and ex vivo experiments. Methods The concentrations of IL-12 were tested by ELISA after mouse dendritic cells (DCs) in vitro were stimulated by one of tested TLR agonists, including poly(I:C), monophosphoryl lipid A (MPLA), resiquimod (R848), cytosine polyguanine-C (CpG-C). The changes in percentage and phenotype of DCs, NK cells and effector T cells in the draining lymph nodes were analyzed by flow cytometry after BALB/c mice were immunized with ovalbumin (OVA) mixed with selected TLR agonist. The serum concentrations of specific anti-OVA IgG2a in the immunized mice were determined by ELISA. Results Only CpG-C and R848 could significantly induce the production of IL-12 from bone marrow-derived DCs in vitro. Among the tested TLR agonists, R848 was the most effective adjuvant in recruiting DCs and NK cells into lymph nodes, inducing the proliferation of CD4+ and CD8+ effector T cells and the production of specific anti-OVA IgG2a in vivo. Conclusion R848 was the most potential Th1-promoting adjuvant among the tested TLR agonists.

Phenotyping and comparing the immune cell populations of free-ranging Atlantic bottlenose dolphins (Tursiops truncatus) and dolphins under human care.

BACKGROUND: Studies suggest that free-ranging bottlenose dolphins exhibit a suppressed immune system because of exposure to contaminants or microorganisms. However, due to a lack of commercially available antibodies specific to marine mammal immune cell surface markers, the research has been indecisive. The purpose of this study was to identify cross-reactive terrestrial-specific antibodies in order to assess the changes in the immune cell populations of dolphins under human care and free-ranging dolphins. The blood and PBMC fraction of blood samples from human care and free-ranging dolphins were characterized by H&E staining of cytospin slides and flow cytometry using a panel of terrestrial-specific antibodies. RESULTS: In this study, we show that out of 65 terrestrial-specific antibodies tested, 11 were cross-reactive and identified dolphin immune cell populations within their peripheral blood. Using these antibodies, we found significant differences in the absolute number of cells expressing specific markers within their lymphocyte and monocyte fractions. Interestingly, the peripheral blood mononuclear cell profile of free-ranging dolphins retained an additional population of cells that divided them into two groups showing a low (<27%) or high (>56%) percentage of smaller cells resembling granulocytes. CONCLUSIONS: We found that the cross-reactive antibodies not only identified specific changes in the immune cells of free-ranging dolphins, but also opened the possibility to investigate the causal relationship between immunosuppression and mortality seen in free-ranging dolphins.

Comparative analysis of dendritic cells and anti-CD3/CD28 expanded regulatory T cells for application in transplantation.

The containment of direct and indirect recognition of donor alloantigens by enhancement of the number and activity of regulatory T cells (Tregs) has been one of the promising approaches to achieve transplant tolerance. Two major methods, dendritic cell (DC) and anti-CD3/CD28 antibodies (Abs) have been introduced for ex vivo expansion of Tregs prior to their adoptive transfer. Here we compared the clinical advantage of using these methods of Tregs expansion by evaluating the nature and ability of expanded Tregs to abolish recipient alloreactive T-cell responses in vitro and in vivo. The ovalbumin (OVA)-specific Tregs isolated from DO11.10 mice were exposed to either Abs or syngeneic DC loaded with OVA peptide in the presence of exogenous IL-2 for a week. Using BABL/c as recipient and C57BL/6 as donor, the suppressive activity of these cells was examined. We found that DCs were much more efficient than Abs in expanding (9-fold versus 4-fold) and maintaining the viability (90% versus 35%) of purified Tregs. Interestingly, the Abs-expanded Tregs superbly contained the alloreactive T-cell proliferation and both Tregs were more suppressive when the Tregs cognate antigen and alloantigens were separately expressed on recipient and donor DCs (HVGD) rather than on recipient DC alone (GVHD). Importantly, however, DC-expanded Tregs maintained stable expression of Foxp3, survived longer and effectively contained the differentiation of alloreactive T cells into IFN-gamma-producing effector cells both in vitro and in vivo. Our data suggests that DC-expanded Tregs provides a clinically advantageous means of preventing unwanted immune reactions to allografts.

B7-1 and B7-2 differentially control peripheral homeostasis of CD4(+)CD25(+)Foxp3(+) regulatory T cells.

Targeting the CD28/B7 interaction remains among the most promising approaches to treat transplant rejection. A drawback to this approach is however the observations of decreased numbers of naturally occurring regulatory T cells (Tregs) in CD28(-) or B7-deficient non-obese diabetic (NOD) mice, cells that maintain immunological self tolerance, prevent autoimmunity and control immune responses to transplants. In this study, therefore, we investigated the relative contributions of B7-1 and B7-2, the only known ligands of CD28, on the thymic development and peripheral homeostasis of Tregs. Our data indicates that the absence of both B7-1 and B7-2 result in a dramatic reduction in the frequencies of Tregs in thymus and peripheral tissues of B7-1/B7-2-deficient mice with no apparent changes in the percentage and distribution of conventional T-cell subsets. In addition, neither B7-1 nor B7-2 expression alone is sufficient for the development and peripheral homeostasis of Tregs. Interestingly, while B7-1 and B7-2 equally contribute to thymic development of Tregs, the significant loss in peripheral homeostasis of Tregs is more evident in the absence of B7-2 as compared to B7-1. Consistent with these results we found that B7-2-deficient DCs are less effective than B7-1-deficient DCs in maintaining Tregs in vitro due to their inability to induce IL-2 production by conventional T cells and sustain Tregs expression of CD25. This study provides the first demonstration of relative roles of B7-1 and B7-2 in Tregs homeostasis and suggests that therapeutic approaches designed to selectively interrupt CD28/B7-2 interaction could indeed have measurable impact on sustaining Tregs homeostasis.

[The research of xeno-lymphocytes apoptosis induced by Sertoli cells isolated by modified methods in vitro].

OBJECTIVE: To modify the isolation and culture method of Sertoli cells and investigate its' effects on xeno-lymphocytes apoptosis. METHODS: Sertoli cells which was isolated from 2 - 4 week-old Sprague Dawley (SD) rats, were successfully prepared by collagenase type V, trypsin and DNase I and then identified by electron microscope. Viability and apoptosis of cultured cells were measured by flow cytometry. The apoptosis rates of Balb/c mouse lymphocytes were examined which were co-cultured with Sertoli cells of SD rats by flow cytometry, too. The expression of FasL, TGF-beta(1) and clusterin on Sertoli cells were detected by immunocytochemistry. RESULTS: In the co-cultured system, Sertoli cells accounted for more than 90%. The viability of Sertoli cells was above 95% and the apoptosis rate was 10.87% +/- 3.87% in this study. The lymphocytes apoptosis ratio was 15.52% +/- 0.17% (P < 0.01). Streptavidin-biotin-peroxidase-complex immunochemistry staining showed that the Sertoli cells could express FasL, TGF-beta(1) and clusterin, respectively. CONCLUSIONS: It indicates that the expression of FasL, TGF-beta(1) on the Sertoli cells might relate to the immune privilege, and it supposed to be benefit for xenotransplantation.

[Establishment of eukaryotic cell line stably expressing soluble human leucocyte antigen G1 by nucleofection].

OBJECTIVE: To establish a eukaryotic cell line that can express soluble human leucocyte antigen G1 (sHLA-G1) stably. METHODS: The recombinant plasmid pcDNA3-sHLA-G1 is transfected by a novel nonviral, electroporation-based gene transfer method termed nucleofection into the host cell lymphoblastoid cell line (LCL) 721.221 which does not express any HLA-classical I molecules. After selection by G418, the cell line stably expressing sHLA-G1 is identified by RT-PCR and Dot-ELISA with HLA-G1 specific monoclonal antibody MEM-G/9. RESULTS: The efficiency of transfection for LCL721. 221 is about 14% by nucleofection. The specific band for sHLA-G1 was found by RT-PCR assay from the transfections and the protein of sHLA-G1 in the supernatant of the transfections was detected by Dot-ELISA assay. Both confirmed that the eukaryotic cell line expressing sHLA-G1 has been established successfully at genic and proteinic levels. CONCLUSION: In this study, the eukaryotic cell line expressing sHLA-G1 have been established successfully by nucleofection.

[Coexpression of human complement regulatory protein genes, CD59 and MCP in transgenic mice protects isolated mouse heart function in ex vivo human plasma perfusion model].

OBJECTIVE: To investigate if the expression of human complement regulatory protein genes in transgenic donor protects against hyperacute rejection (HAR) in the recipient. METHODS: Kunming mice were transfected with 2 human complement regulatory protein genes, CD59 and MCP, so as to establish a transgenic hCD59/hMCP mouse model. Eight hCD59 expression mice, 11 hMCP expression mice, and 8 hCD59/hMCP expression mice were used as experimental groups, and 10 transgenic negative littermates were used as control group. The hearts of the mice were taken out to be perfused with 10% pooled human blood of B type. During the perfusion electrocardiography was carried out to observe the beating time. After the hearts stopped beating, immunofluorescence staining and immunohistochemistry were used to detect the deposition of complements C(9) and C(3c) in the heart tissue. RESULTS: The mean heart beating time was 138 +/- 25 minutes in the hCD59/hMCP expression group, 78 +/- 27 minutes in the hCD59 expression group, 43 +/- 21 minutes in the hMCP group, and 20 +/- 12 minutes in the wild type nontransgenic control group (all P < 0.01, Dennett's T test for all 3 other groups relative to the hCD59/hMCP group). Deposition of the complement C(9) and that of C(3c) were not found in the hearts of the hCD59/hMCP group, however, could be found in the hearts of the 2 monotransgenic groups and nontransgenic group. CONCLUSION: The coexpression of human complement regulatory protein genes, CD59 and MCP in the xenografts effectively inhibits the complement of xenograft-mediated HAR.