NK Group Research Articles

Longitudinal Analysis of Renal Function Changes in Elderly Populations: Health Status Evaluation and Risk Factor Assessment.

This study aims to investigate GFR decline in elderly subjects with varying physical conditions and analyze key risk factors impacting renal function changes. We obtained data from patients between 2017 and 2019, and matched healthy elderly subjects based on gender and age. Data collected for all subjects included annual measurements of fast blood glucose (GLU), glycated hemoglobin (HbA1c), low-density lipoprotein cholesterol (LDL-c), blood albumin (ALB), blood uric acid (UA), urine protein (UP), and systolic blood pressure (SBP). Additionally, information on coexisting diseases was gathered. The Full Age Spectrum (FAS) equation was used to calculate eGFR. A total of 162 patients with complete 3-year renal dynamic imaging were included, including 84 patients in the kidney disease group (K group) and 78 patients in the non-kidney disease group (NK group). Ninety individuals were selected as the healthy group (H group). The annual decline rate in the K group was the fastest, which exceeded 5mL/min/1.73m2 (P < 0.05). Group (K group: β=-40.31, P<0.001; NK group: β=-26.96, P<0.001), ALB (β=-0.38, P=0.038) and HbA1c (β=1.36, P=0.029) had a significant negative impact on the eGFR changes. For participants who had negative proteinuria: K group had the most significant annual eGFR decline. The presence of kidney disease, along with proteinuria nor not, can lead to a marked acceleration in kidney function decline in elderly. We categorize elderly individuals with an annual eGFR decline of more than 5 mL/min/1.73m2 as the "kidney accelerated aging" population.

Complete Genomic Assembly of Mauritian Cynomolgus Macaque Killer Ig-like Receptor and Natural Killer Group 2 Haplotypes.

Mauritian-origin cynomolgus macaques (MCMs) serve as a powerful nonhuman primate model in biomedical research due to their unique genetic homogeneity, which simplifies experimental designs. Despite their extensive use, a comprehensive understanding of crucial immune-regulating gene families, particularly killer Ig-like receptors (KIR) and NK group 2 (NKG2), has been hindered by the lack of detailed genomic reference assemblies. In this study, we employ advanced long-read sequencing techniques to completely assemble eight KIR and seven NKG2 genomic haplotypes, providing an extensive insight into the structural and allelic diversity of these immunoregulatory gene clusters. Leveraging these genomic resources, we prototype a strategy for genotyping KIR and NKG2 using short-read, whole-exome capture data, illustrating the potential for cost-effective multilocus genotyping at colony scale. These results mark a significant enhancement for biomedical research in MCMs and underscore the feasibility of broad-scale genetic investigations.

Cutting Edge: PDGF-DD Binding to NKp44 Costimulates TLR9 Signaling and Proinflammatory Cytokine Secretion in Human Plasmacytoid Dendritic Cells.

NKp44 is a human receptor originally found on activated NK cells, group 1 and group 3 innate lymphoid cells that binds dimers of platelet-derived growth factor D (PDGF-DD). NKp44 is also expressed on tissue plasmacytoid dendritic cells (PDCs), but NKp44-PDGF-DD interaction on PDCs remains unstudied. Engagement of NKp44 with PDGF-DD invitro enhanced PDC secretion of IFN-α, TNF, and IL-6 in response to the TLR9 ligand CpG-ODN, but not TLR7/8 ligands. In tissues, PDCs were found in close contact with PDGF-DD-expressing cells in the high endothelial venules and epithelium of tonsils, melanomas, and skin lesions infected with Molluscum contagiosum. Recombinant PDGF-DD enhanced the serum IFN-α response to systemic HSV-1 infection in a humanized mouse model. We conclude that NKp44 integrates with TLR9 signaling to enhance PDC cytokine production. These findings may have bearings for immune responses to TLR9-based adjuvants, therapy for tumors expressing PDGF-DD, and infections with DNA viruses that induce PDGF-DD expression to enhance viral spread.

Analysis of Butyrophilin-Mediated Activation of γδ T Cells from Human Spleen.

There is considerable interest in therapeutically engaging human γδ T cells. However, due to the unique TCRs of human γδ T cells, studies from animal models have provided limited directly applicable insights, and human γδ T cells from key immunological tissues remain poorly characterized. In this study, we investigated γδ T cells from human spleen tissue. Compared to blood, where Vδ2+Vγ9+ T cells are the dominant subset, splenic γδ T cells included a variety of TCR types, with Vδ1+ T cells typically being the most frequent. Intracellular cytokine staining revealed that IFN-γ was produced by a substantial fraction of splenic γδ T cells, IL-17A by a small fraction, and IL-4 was minimal. Primary splenic γδ T cells frequently expressed NKG2D (NK group 2 member D) and CD16, whereas expression of DNAM-1 (DNAX accessory molecule 1), CD28, PD-1, TIGIT, and CD94 varied according to subset, and there was generally little expression of natural cytotoxicity receptors, TIM-3, LAG-3, or killer Ig-like receptors. In vitro expansion was associated with marked changes in expression of these activating and inhibitory receptors. Analysis of functional responses of spleen-derived Vδ2+Vγ9+, Vδ1+Vγ9+, and Vδ1+Vγ9- T cell lines to recombinant butyrophilin BTN2A1 and BTN3A1 demonstrated that both Vδ2+Vγ9+ and Vδ1+Vγ9+ T cells were capable of responding to the extracellular domain of BTN2A1, whereas the addition of BTN3A1 only markedly enhanced the responses of Vδ2+Vγ9+ T cells. Conversely, Vδ1+Vγ9+ T cells appeared more responsive than Vδ2+Vγ9+ T cells to TCR-independent NKG2D stimulation. Thus, despite shared recognition of BTN2A1, differential effects of BTN3A1 and coreceptors may segregate target cell responses of Vδ2+Vγ9+ and Vδ1+Vγ9+ T cells.

Assessing the efficacy of four methods established by four parameters in ICL size selection and relevant influencing factors: a prospective cohort study.

To compare the efficacy and relevant influencing factors of four ICL size selection methods established by four different parameters. This prospective study included 60 patients (120 eyes) who underwent bilateral ICL implantation. Patients were equally divided into four groups, and each group used the Parkhurst nomogram based on sulcus-to-sulcus (STS), the manufacturer's Online Calculation & Ordering System (OCOS) nomogram based on white-to-white (WTW), the KS formula based on angle-to-angle (ATA) and the NK formula based on anterior chamber width (ACW) to determine the ICL size. Recorded the vault one month after operation and compared the consistency between STS and WTW, ATA and ACW and their effects on the vault of different groups. The Parkhurst nomogram, OCOS nomogram, KS formula and NK formula determined 86.7%, 70.0%, 83.3% and 66.7% of properly sized ICL, respectively. STS and ATA were correlated (P < 0.05). The mean difference between the STS and WTW, ATA and ACW was -0.37 ± 0.62mm, -0.42 ± 0.53mm and -0.44 ± 0.52mm, respectively. The vault in the OCOS group was negatively correlated with △STS-WTW, and the vault in the NK group was negatively correlated with △STS-WTW, △STS-ATA and △STA-ACW. The vault in the Parkhurst group and KS group was not affected by anterior segment biometry variables. ATA can be served as an alternative parameter to STS, and STS-based Parkhurst nomogram and ATA-based KS formula determined the most appropriate ICL size. When using OCOS nomogram and NK formula to select ICL size, postoperative abnormal vault was associated with a larger difference between STS and other anterior segment parameters.

A Crosstalk Between Castration-Resistant Prostate Cancer Cells, M2 Macrophages, and NK Cells: Role of the ATM-PI3K/AKT-PD-L1 Pathway

ABSTRACT Castration-resistant prostate cancer (CRPC) in males is associated with a poor prognosis and a higher risk of treatment-related adverse effects, with high mortality among cancers globally. It is thus imperative to explore novel potential molecules with dual therapeutic and biomarker functions. Based on the recent research findings, the expression levels of ataxia telangiectasia mutant kinase (ATM) in prostate cancer (PC) tissues collected from CRPC patients were higher than hormone-dependent PC patients. Using CRPC cell lines (C4–2 and CWR22Rv1), the transwell chamber experiments revealed ATM promoted macrophage recruitment in CRPC cells in vitro via C-X-C motif chemokine ligand 12 (CXCL12). Further in vitro investigations demonstrated that polarized macrophages prevented NK cell recruitment and reduced the immunocidal activity of NK cells against CRPC cell lines. Moreover, ATM boosted programmed death receptor ligand 1 (PD-L1) expression while inhibiting NK group 2D (NKG2D) ligand expression in selected cell lines via PI3K/AKT signaling pathway. The in vivo investigations revealed ATM induced proliferation of CRPC and macrophage recruitment, while the NK cell recruitment was found to suppress ATM expression and CRPC proliferation. In conclusion, it could be demonstrated that inhibiting ATM increased the susceptibility of CRPC to NK cell inhibitors by dampening the CXCL12 and PI3K/AKT-PD-L1 pathways, thereby offering a novel and individualized treatment protocol for treating CRPC.

The effectiveness of a novel treatment of TIM-3(-) NK cells infusion in murine models of immune-mediated bone marrow failure.

T-cell immunoglobulin and mucin-containing domain (TIM)-3 exerts its inhibitory effect on NK cells and participates in the immune pathogenesis of SAA. In this study, we aimed to explore a novel treatment method of TIM-3(+) NK or TIM-3(-) NK cell infusion in combination with immunosuppressive therapy for bone marrow failure (BMF)/aplastic anemia (AA) mice. BMF/AA mouse model was constructed. The TIM-3 expression and functional molecules on TIM-3(+) and TIM-3(-) NK cells of the BMF group, total body irradiation (TBI) group, and normal control (NC) group mice were detected by flow cytometry. After treatment, the general condition, whole blood cell and bone marrow cell (BMC) count, and immune condition of mice from each group were compared. TIM-3 expression in the peripheral blood NK cells of BMF mice was significantly lower than that of the TBI and NC group mice. TIM-3(-) NK cells expressed more NKG2D receptors than TIM-3(+) NK cells. The levels of P-Akt and PI3K in TIM-3(-) NK cells were higher than those in TIM-3(+) NK cells. On the 17th day after BMF induction, the weight, peripheral whole blood cell count, and BMC count of BMF mice decreased significantly compared with that of the NC group mice. The therapeutic effect in the TIM-3(-) NK cell treatment group was better than that in the TIM-3(+) NK cell treatment and CsA treatment groups. Concurrently, the ratio of CD4+ T and CD8+ T cells of BMF mice was significantly lower than that of the NC group mice. The therapeutic effect in CsA + TIM-3(-) NK group was more significant than that of the CsA treatment and the CsA + TIM-3(+) NK groups. In this study, we found that the general condition, peripheral whole blood cell and BMC count, and immune status of BMF mice improved significantly after CsA + TIM-3(-) NK cell treatment. These results may provide further insights into the immune pathogenesis of SAA and novel therapeutic ideas for improving SAA treatment.

Predicting post-laminoplasty kyphosis in cervical spondylotic myelopathy patients without preoperative kyphosis: a retrospective study

BackgroundThis study aimed to determine potential risk factors for post-laminoplasty kyphosis and the effect of postoperative kyphosis on neurologic function recovery.MethodsA total of 266 patients with cervical spondylotic myelopathy (CSM) underwent traditional cervical laminoplasty with a minimum of a 12-month follow-up period. The patients were divided into non-kyphosis (NK group) and kyphosis (K group) groups based on the postoperative C2-7 Cobb angle. Clinical and radiological measurements were collected preoperatively and at the final follow-up.ResultsOf the 266 patients, 26 (9.77%) developed postoperative kyphosis at the final follow-up. The postoperative Japanese Orthopedic Association score did not differ significantly between the NK and K groups (P > 0.05). The postoperative numeric rating scale (NRS) also showed no significant difference between the NK and K groups; however, postoperative NRS improved better than the preoperative values in the NK group (P < 0.001). Multivariate analysis revealed that the preoperative C2-7 extension Cobb angle and C2-7 Cobb angle were independent predictors of post-laminoplasty kyphosis. Cut-off values for predicting postoperative kyphosis were a C2-7 extension Cobb angle of 18.00° and a C2-7 Cobb angle of 9.30°.ConclusionsLow preoperative C2-7 extension Cobb angle and C2-7 Cobb angle may be associated with post-laminoplasty kyphosis in CSM patients without preoperative kyphosis. The cut-off value of the C2-7 extension Cobb angle and C2-7 Cobb angle were 18.00° and 9.30°, respectively.

Orally delivered Bacillus subtilis expressing chicken NK-2 peptide stabilizes gut microbiota and enhances intestinal health and local immunity in coccidiosis-infected broiler chickens

We recently reported a stable Bacillus subtilis-carrying chicken NK-lysin peptide (B. subtilis-cNK-2) as an effective oral delivery system of an antimicrobial peptide to the gut with therapeutic effect against Eimeria parasites in broiler chickens. To further investigate the effects of a higher dose of an oral B. subtilis-cNK-2 treatment on coccidiosis, intestinal health, and gut microbiota composition, 100 (14-day-old) broiler chickens were allocated into 4 treatment groups in a randomized design: 1) uninfected control (CON), 2) infected control without B. subtilis (NC), 3) B. subtilis with empty vector (EV), and 4) B. subtilis with cNK-2 (NK). All chickens, except the CON group, were infected with 5,000 sporulated Eimeria acervulina (E. acervulina) oocysts on d 15. Chickens given B. subtilis (EV and NK) were orally gavaged (1×1012 cfu/mL) daily from d 14 to 18. Growth performances were measured on d 6, 9, and 13 postinfection (dpi). Spleen and duodenal samples were collected on 6 dpi to assess the gut microbiota, and gene expressions of gut integrity and local inflammation makers. Fecal samples were collected from 6 to 9 dpi to enumerate oocyst shedding. Blood samples were collected on 13 dpi to measure the serum 3-1E antibody levels. Chickens in the NK group showed significantly improved (P < 0.05) growth performance, gut integrity, reduced fecal oocyst shedding and mucosal immunity compared to NC. Interestingly, there was a distinct shift in the gut microbiota profile in the NK group compared to that of NC and EV chickens. Upon challenge with E. acervulina, the percentage of Firmicutes was reduced and that of Cyanobacteria increased. In NK chickens, however, the ratio between Firmicutes and Cyanobacteria was not affected and was similar to that of CON chickens. Taken together, NK treatment restored dysbiosis incurred by E. acervulina infection and showed the general protective effects of orally delivered B. subtilis-cNK-2 on coccidiosis infection. This includes reduction of fecal oocyst shedding, enhancement of local protective immunity, and maintenance of gut microbiota homeostasis in broiler chickens.

Cutting Edge: Bispecific γδ T Cell Engager Containing Heterodimeric BTN2A1 and BTN3A1 Promotes Targeted Activation of Vγ9Vδ2+ T Cells in the Presence of Costimulation by CD28 or NKG2D

Vγ9Vδ2+ T cell-targeted immunotherapy is of interest to harness its MHC-independent cytotoxic potential against a variety of cancers. Recent studies have identified heterodimeric butyrophilin (BTN) 2A1 and BTN3A1 as the molecular entity providing "signal 1" to the Vγ9Vδ2 TCR, but "signal 2" costimulatory requirements remain unclear. Using a tumor cell-free assay, we demonstrated that a BTN2A1/3A1 heterodimeric fusion protein activated human Vγ9Vδ2+ T cells, but only in the presence of costimulatory signal via CD28 or NK group 2 member D. Nonetheless, addition of a bispecific γδ T cell engager BTN2A1/3A1-Fc-CD19scFv alone enhanced granzyme B-mediated killing of human CD19+ lymphoma cells when cocultured with Vγ9Vδ2+ T cells, suggesting expression of costimulatory ligand(s) on tumor cells is sufficient to satisfy the "signal 2" requirement. These results highlight the parallels of signal 1 and signal 2 requirements in αβ and γδ T cell activation and demonstrate the utility of heterodimeric BTNs to promote targeted activation of γδ T cells.

Low potassium disrupt intestinal barrier and result in bacterial translocation

BackgroundBacterial translocation was observed in critical illness and patients with chronic diseases such as liver cirrhosis and chronic kidney disease (CKD). Hypokalemia is a common complication in these diseases. Whether low potassium diet may increase intestinal permeability and result in bacterial translocation lack of evidence. The present study was aimed to investigate the potential effects of LK on intestinal permeability.MethodsGrade 8-week-old male Bal B/C mice were randomly placed either on a normal potassium (NK) mouse chow or a low potassium (LK) diet for 28 days. Intestinal permeability and expression of tight junction proteins were compared between the two groups.ResultsCompared with the NK group, the mice in LK group had significantly lower serum potassium level, increased levels of plasmas endotoxin and plasma d-lactate. The bacterial translocation was higher and in occurred mainly in mesenteric lymph nodes (MLN), liver and spleen. The pathologic change of small intestine was obvious with thinner villus lamina propria, shorter crypt depth and thinner intestinal wall. Slight increases in the expression of proteins and mRNA levels of both claudin-1 and claudin-2 were observed in LK group.ConclusionsLow potassium diet could increase intestinal permeability and thereby lead to bacterial translocation, which was suspected to result from impaired intestinal epithelial barrier and biological barrier.

Peroxisome Proliferator-Activated Receptor Gene Knockout Promotes Podocyte Injury in Diabetic Mice

Objective To investigate the effects of peroxisome proliferator-activated receptor (PPARγ) expression on renal podocyte in diabetic mice by conditionally knockout mouse PPARγ gene. Methods Wild-type C57BL mice and PPARγ gene knockout mice were used as research objects to establish the diabetic mouse model, which was divided into normal control group (NC group), normal glucose PPARγ gene knockout group (NK group), diabetic wild-type group (DM group), and diabetic PPARγ gene knockout group (DK group), with 8 mice in each group. After 16 weeks, the mice were sacrificed for renal tissue collection. Morphological changes of renal tissue were observed by HE and Masson staining, and ultrastructure of renal tissue was observed by transmission electron microscope. Protein expressions of PPARγ, podocin, nephrin, collagen IV, and fibronectin (FN) in renal tissues were detected by immunohistochemistry and Western blot, and mRNA changes of PPARγ, podocin, and nephrin in renal tissues were detected by qRT-PCR. Results Compared with the NC group, the protein and mRNA expressions of PPARγ, podocin, and nephrin decreased in the kidney tissue of mice in the DM group, while the protein expressions of collagen IV and FN increased. The expression of various proteins in kidney tissues of the DK group was consistent with that of the DM group, and the difference was more obvious. The expression of PPARγ protein and mRNA decreased in the NK group, while the expression of podocin, nephrin protein and mRNA, collagen IV, and FN protein showed no significant difference. Conclusion In diabetic renal tissue, the loss of PPARγ can aggravate podocellular damage and thus promote the occurrence of diabetic renal fibrosis. Increasing the expression of PPARγ may effectively relieve renal podocyte impairment in diabetic patients, which can be used for the treatment of diabetic nephropathy.

Combined treatment with anti-PSMA CAR NK-92 cell and anti-PD-L1 monoclonal antibody enhances the antitumour efficacy against castration-resistant prostate cancer.

BackgroundThe chimeric antigen receptor NK‐92 (CAR NK‐92) cell targeting the prostate‐specific membrane antigen (PSMA) has shown antitumour effects in castration‐resistant prostate cancer (CRPC). However, the expression changes of programmed death ligand 1 (PD‐L1) and its mechanisms on CAR NK‐92 and CRPC cells and the effect of the anti‐PD‐L1 monoclonal antibody (mAb) on PD‐L1 expressed on CAR NK‐92 cells remain unknown.MethodsHuman dendritic cells and CD8+ T cells were acquired from blood samples of healthy donors and cocultured with C4‐2 cells. Changes in PD‐L1 expression were detected by flow cytometry. Differential gene expressions were investigated by RNA sequence analysis, while the regulation of PD‐L1 molecular signaling was explored using western blotting. In vitro cytotoxicity was evaluated using the Cell Counting Kit‐8 assay and the bioluminescent intensity (BLI) of green fluorescent protein‐labelled C4‐2 cells. CRPC growth in vivo was monitored using callipers and BLI in male NOD/SCID mice subcutaneously injected with C4‐2 cells and treated intravenously with anti‐PD‐L1/PD‐1 mAb, CAR NK‐92 or cocultured CD8+ T cells.ResultsSignificantly upregulated expression of PD‐L1k was observed in cocultured C4‐2 and CAR NK‐92 cells. In addition, upregulation of PD‐L1 expression was dependent on interferon‐γ in C4‐2 cells, while it was dependent on direct cell‐to‐cell interaction via the NK group 2 member D/ phosphatidylinositol 3‐kinase/AKT pathway in CAR NK‐92 cells. The anti‐PD‐L1 mAb directly acted on PD‐L1 expressed on CAR NK‐92 cells and augmented the cytotoxicity of CAR NK‐92 cells against C4‐2 and CRPC cells from one patient in vitro. Anti‐PD‐L1 mAb significantly enhanced the antitumour effect of CAR NK‐92 cells against CRPC cells in vivo when compared to treatment with CAR NK‐92 cells or combined with anti‐PD‐1 mAb in the absence or presence of cocultured CD8+ T cells.ConclusionCombined treatment with CAR NK‐92 and anti‐PD‐L1 mAb improved the antitumour efficacy against CRPC, which is of extraordinary translational value in the clinical treatment of CRPC.

Effect of Lanthanum Zirconate Ceramic on the Dynamics of Hematological Parameters and the Bone Remodeling Markers: Experimental Study

Background. Zirconium oxide is actively used in medicine; however, research is underway to improve mechanical characteristics and biointegration. One of the promising areas is the study of materials based on lanthanum zirconate (LZ).&#x0D; The study aimed to examine the effect of a new ceramic material based on LZ on the dynamics of hematological parameters and markers of bone tissue remodeling after intramedullary osteosynthesis (IO) of a hip fracture with LZ rods.&#x0D; Material and Methods. The ceramic material La1.95Ca0.05Zr2O7 was used. The experiment was conducted in guinea pigs, which were divided into four groups: main group, modeling of a hip fracture (IO of the fracture with LZ rod, n = 9); comparison group, modeling of a hip fracture (IO of a fracture with a rod from b-tricalcium phosphate [TCP]; n = 9); control (C) group, modeling of a hip fracture without IO (n= 9); and native control (NC) group. Animals were withdrawn from the experiment before surgery (NC) and at 4, 10, and 25 weeks after surgery (n=3 for each time point). Hematological parameters, i.e., a tartrate-resistant acid phosphatase (TRAP) as an osteoresorption marker and osteocalcin (OC) as an osteogenesis marker, were determined.&#x0D; Results. The red blood cell counts in all groups of the operated animals at 4, 10, and 25 weeks after surgery were not significantly different from the NK group. A significantly higher level of leukocytes in comparison with other groups was observed in the control group 10 weeks after surgery (p = 0.044), which was explained by the absence of fracture synthesis. The platelet level in all groups of the operated animals during the study period was not significantly different from the NK group. The TRAP activity in the LZ and TCP groups had maximum values after 4 weeks, and the OC level reached the maximum by 10 weeks after the operation without significant differences between the LZ and TCP groups of animals.&#x0D; Conclusion. The study of the main hematological parameters did not reveal a negative effect of LZ on the experimental animals. A positive effect of this material on bone tissue remodeling was found. A new ceramic material based on LC appears to be promising for use in traumatology and orthopedics.

Compound green tea (CGT) regulates lipid metabolism in high-fat diet induced mice.

This work aims to study the effect of compound green tea (CGT) on liver lipid metabolism in mice based on metabolomics of liquid chromatography-mass spectrometry (LC-MS), and preliminarily identify potential biomarkers and pathways of action by using a metabonomic network database to explore the lipid-lowering effect of CGT. In this study, forty mice were randomly divided into four groups: compound tea treatment group (DH), high-fat model control group (NK), normal control group (CK) and positive drug group (YK). After a month of different interventions, the mice were weighed and the blood lipid indexes were detected. In addition, differential liver metabolites were monitored by using LC-MS. The results showed that CGT and positive drug treatment were able to decrease body weight, liver coefficient, TC, TG and LDL levels of obese mice, while increasing HDL levels. Among the 110 compounds obtained, 54 metabolites were significantly altered in the four comparisons. More importantly, 15 remarkably downregulated metabolites involved in Lysopc 16:1, Lysopc 18:1, and Lysopc 18:2 were found in the DH group when the mice were treated with CGT; meanwhile, the positive drug Xuezhikang was able to significantly downregulate 14 compounds, including (±)18-HEPE, and 6 keto-PGF1α, compared with the NK group. Besides, KEGG enrichment analysis also revealed the important metabolic pathways, such as linoleic acid metabolism, Biosynthesis of unsaturated fatty acids, and α-linolenic acid metabolism, were related to fatty acid metabolism. These results suggested that CGT could regulate the lipid metabolism in the liver of hyperlipidemia mice, and may regulate 54 potential biomarkers in mice through a related metabolic pathway to make them return to a normal state and improve the disorder of lipid metabolism.

Comparison of Ketamine Nebulisation with Ketamine Gargle in Attenuating Post Operative Sore Throat Following General Anaesthesia with Endotracheal Intubation

Background &amp; Objective: In the post operative period, Sore throat is a common morbidity following tracheal intubation. The aim was to compare ketamine nebulisation with ketamine gargle to see the effectiveness of nebulisation over gargle in reducing the incidence and severity of post operative sore throat (POST).&#x0D; Methods: In this prospective, randomized study 80 patients between 18-64 year, of either sex belonging to ASA I/II, scheduled for elective surgery under general anaesthesia, were assigned into two groups of 40 patients each. Group GK received preservative free ketamine 50mg in 29 ml of saline, gargled for 30 seconds and Group NK received ketamine 50 mg in 4ml of normal saline via nebulisation for 15 min. General anaesthesia was induced 5 min later in both the groups. Postoperatively sore throat was assessed at 0, 2, 4, 24 hrs using four point scale (0-3).&#x0D; Results: The incidence of POST at 0, 2, 4, 24 hrs was 27%, 25%, 20% and 17% respectively in GK group and 20%, 17.5%, 12.5% and 7.5% in NK group respectively with no statistical difference between the two.. None of the patients in either group experienced severe sore throat.&#x0D; Conclusion: Ketamine nebulisation seemed to represent an effective alternative to ketamine gargle for reliving Post Operative sore throat.

The Significance of Chromosome 11 Abnormalities in Patients with AML

Introduction: At the time of diagnosis AML the finding of KMT2A gene rearrangement qualifies patient to the group with adverse cytogenetic risk according to ELN 2017. In many hematooncological diseases including AML abnormalities concerning another regions of chromosome 11 are detected. However, their significance for outcome, CR rate, OS is still unknown. The main aim of this study was analysis of chromosome 11 abnormalities in AML patients, focusing on these without KMT2A gene rearrangement and its influence on patients outcome.Methods: Karyotype analysis from bone marrow aspirate was performed for patients with chromosome 11 abnormalities (abn11, n=36) and compared to two groups: with complex (CK, n=20) and normal karyotype (NK, n=44). The basic clinical and laboratory data including cytogenetic (karyotype and FISH method) and presence of TP53 deletion were analyzed to characterize in detail the group of patients with chromosome 11 abnormalities.Patients were treated with intensive chemotherapy, in 33% of them allogeneic cell transplantation (alloHCT) was performed.Results: In patients with abn11 the most frequent type of aberration was translocation, 11q23(+)/KMT2A(-) and 11q10-25 as a main region involved in aberrations, with minimal narrowing of region to 11q22. In patients with abn11 and translocation (excluding t(9;11)) there was higher frequency occurrence of 1 to 2 additional abnormalities (p=0.033) and lower frequency of complex karyotype existence (p=0.026). The gain of genetic material from chromosome 11 was observed in 33% of patients with abn11 group and the minimal amplified region was 11q22. Furthermore, in patients with amplification of chromosome 11 region, different additional cytogenetic abnormalities (such as monosomy 5 (p=0.045) and 18 (p<0.001)) were found in comparison to patients with other abnormalities of chromosome 11. Additional KMT2Agene copies were often observed in patients with 11q10-25 (p=0.013) and 11pter-qter (p=0.022) region involvement. The lowest frequency of TP53 deletion was in NK group (p=0.002) but comparable in patients with CK and abn11 without impact on OS and CR. The FISH analysis did not revealed KMT2A gene rearrangement in group with normal karyotype. There was particular attention to 11q23 region in patients with AML which enabled abnormality detection in GTG analysis, furthermore KMT2A rearrangement was confirmed or excluded by FISH method. In few cases FISH analysis allowed verification of KMT2A gene status in case of atypical hybrydization pattern occurrence.Leukocyte counts (p<0.001) and type of AML (de novo, secondary, therapy-related, p=0.022) differs in patients with abn11 in comparison to remained studied subgroups (Table 1.). The lowest blast percentage was observed in patients with 11p rearrangement (p=0.045). There were no significant differences in OS between all 3 groups of patients. CR was achieved in 15% in a CK group and it was significantly lower than in NK and abn11 group (p<0.001). Furthermore alloHCT improve OS in all treated patients (p<0.001). Additional genetic material from chromosome 11 occurrence had negative impact on OS (p<0.001) and CR (p=0.007). Patients with abn11 who achieved CR had lower median age (p=0.01) compared to these without CR. The lowest percentage of CR in abn11 group was observed in patients with additional KMT2A gene copies (p=0.008), 11q10-25 region involvement (p=0.036), HSR (homogeneously staining region) and aneuploidy (p=0.04).Conclusions: In summary, the most frequent region involved in chromosome 11 aberration was 11q22. Additional material derived from chromosome 11 had negative impact on CR and OS. Cytogenetic and molecular analysis in patients with AML are needed to detect most of abn11. In the era of targeted therapies their identification is very important to qualify the patient for more effective treatment. Both cytogenetic and molecular techniques widespread knowledge about cancer biology, however, classic cytogenetic is still “gold standard” and crucial method in AML diagnosis. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Response to Erythropoietin in Different Karyotipic Subsets of Myelodysplastic Syndromes (MDS) and MDS with Sideroblasts: Multicentric Real-Life Experience from the Grom-L Cooperative Group

Background Erythropoietin (EPO) has been widely employed as first-line approach in the treatment of all patients with low-risk Myelodysplastic Syndromes (MDS) and anemia, with response rates ranging from 30 to 60%. Among all low-risk MDS patients, however, there is increasing attention to recognize different subsets characterized by specific karyotipic (del5q-, del20q-), molecular or morphologic (ring sideroblasts) features and, in some cases, by very effective specific treatments (lenalidomide, luspatercept). The role of EPO treatment in these subsets has been very rarely evaluated and is still a matter of debat.Methods To address this issue, the Gruppo Laziale Mielodisplasie (GROM-L) revised retrospectively 316 MDS patients treated with EPO from 1/2002 to 12/2010 by 11 Hematological Centers (5 university hospitals and 6 community-based hospitals) in the metropolitan area of Rome and belonging to 4 different subsets: 82 patients with del5q- isolated or associated to 1 other karyotypic alteration (del5 group), 18 patients with isolated del20q- (del20 group), 41 patients with ring sideroblasts > 15% (RS group) and 175 patients with normal karyotype (NK group) as control group. The main features of these 4 groups are reported in Table 1.Results In Table 2are reported median intervals from diagnosis to EPO start, types of EPO and initial EPO dosages as well as rates of erythroid response (Hb increase > 1.5 g/dl and/or disappearance of transfusion requirement) in the 4 MDS groups. In particular, compared with NK group as control cohort, del5q group had a significant lower rate of erythroid response (p=0.001), while del20q and RS groups showed a similar rate (p=0.431 and p=0.425, respectively). The number of patients with a relapse during EPO treatment was significantly lower in the NK group compared to the other 3 groups, as shown in Table 2 (p=0.005). Median overall survival (OS) from EPO start in the 4 groups is reported in Table 2 while cumulative OS is shown in Figure 1: compared with NK group, only RS group had a better cumulative OS (p=0.026), while both del5q and del20q groups had similar OS (p=0.455 and p=0.367, respectively).Conclusions Our real-life data from a single homogeneous geographic area outline that EPO treatment is effective in all subsets of low-risk MDS considered in this study: the del5q- group had a lower rate of erythroid response, which in any case exceeded 40% of patients. These data, coupled with the excellent toxicity profile of EPO, seem reinforce this treatment as 1st line approach also in those subsets of MDS with specific and very effective (but also quite toxic) new drugs [Display omitted] DisclosuresBreccia:Incyte: Consultancy; Bristol Myers Squibb: Consultancy; Pfizer: Consultancy; Novartis: Consultancy.

A 11-Year Clinical Summary of DC - CIK / NK Cell Immunotherapy for 152 Patients with Acute Myeloid Leukemia

Objective: To assess the efficacy of immunotherapy based on autologous dendritic cells-cytokine induced killer cells (DC-CIK) and allogeneic natural killer cells (NK) in treating low-risk and intermediate-risk acute myeloid leukemia (AML). The study was conducted over a period of 11 years (April 1, 2006 to April 1, 2017) across the Beijing Ludaopei hospital systems.Methods: DC-CIK cells were derived by culturing patients' peripheral blood mononuclear cells (PBMCs) in vitro with cytokines for 8-12 days. To generate NK cells, allogenic PBMCs were cultured for 3 to 6 days. After patients completed intensified chemotherapy consisting of 4 cycles of high-dose Ara-C within 6 months, DC-CIK or NK cells were infused and given once every 3 months for 2-4 cycles along with chemotherapy of Fludarabine, Cyclophosphamide and Ara-C given few days prior to each cycle of immunotherapy.Results: From April 1, 2006 to April 1, 2017, a total 152 patients in the low and intermediate risk groups of AML (except APL) underwent combined immunotherapy and chemotherapy in our center. Overall survival (OS) rate was 82% and disease-free survival (DFS) rate was 67% for the cohort. Beginning in June 2012, the low and intermediate risk groups were further stratified into very-good risk, good risk, and intermediate risk groups according to the specific AML related gene mutations. The OS rate of the 2012-2017 group was significantly better than that of the 2006-2011 group (91.7% vs. 71.6% p= 0.003). For patients treated prior to 2012, there were no significant differences between the low and intermediate risk group in OS (72.2% vs 71.4%) and DFS (72.2% vs 61.2%). From 2012 to 2017, the OS rates were 94.4%, 86.3%, and 93.3%, and the DFS rates were 83.3%, 81.8%, and 62.2%, (p=0.15) for the very-good risk, good-risk & intermediate risk groups, respectively, again suggesting no statistical differences among the groups. Side effects were mild with some fever, chills and fatigue.Patients who were treated with 2-4 cycles of immunotherapy were further divided into the DC-CIK alone group and the DC-CIK alternating with NK group. Importantly, the OS and DFS rates of 67 patients in the DC-CIK alternating with NK group were significantly better than those of 53 patients in the DC-CIK alone group (OS 95.5% vs 71.4%, p=0.0003), (DFS 85% vs 63.5%, p =0.0099). .Twenty-nine of 48 relapsed patients from both groups underwent allo-hematopoietic stem cell transplantation (allo-HSCT). The long-term OS after HSCT was 65.5%, closed to the OS of AML patients who transplanted in their first complete remission (CR) in our center. OS of 12 patients transplanted after 2012 was significantly better than that of 17 patients transplanted before 2012 (88.2% vs. 33.3%, p = 0.015). This because that after 2012, all patients have been checked minimal residual disease (MRD) by flow cytometry. Once patient was found to have relapse with MRD,immediately pursued HSCT while some patients transplanted prior to 2012 had morphological relapse.Conclusion: When combined with chemotherapy, DC-CIK/NK cells-based immunotherapy significantly improves the long-term OS and DFS rates of good and intermediate risk groups of AML. Auto DC-CIK alternating with allo NK is superior to auto DC-CIK alone. Relapsed patients could be safely and effectively treated with HSCT. If relapse patients with only positive MRD, then the outcome of HSCT will be comparable to those who received the HSCT during their first CR.Figure 1. OS of 152 patients treated with DC-CIK/NK immunotherapy from 2006 to 2017.Figure 2, OS of 67 patients treated from 2006 to 2012 vs 85 patients from 2012 to 2017 (71.6% vs 91.7%, p= 0.003).Figure 3, OS & DFS of DC-CIK+NK vs DC-CIK treated groups from 2012 to 2017 (p= 0.0003, p= 0.0099) [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Intraoperative Nice knots assistance for reduction in displaced comminuted clavicle fractures

PurposeThe Nice knots have been widely used in orthopedic surgeries to fix torn soft tissue and fracture in recent years. The study aims to investigate the clinical efficacy and prognosis of intraoperative and postoperative Nice Knots-assisted reduction in the treatment of displaced comminuted clavicle fracture.MethodsFrom Jan 2014 to Dec 2019, 75 patients diagnosed with unilateral closed displaced comminuted clavicle fracture were treated with open reduction and internal fixation (ORIF) in this study. Nice knot group (the NK group) included 38 patients and the other 37 patients were in the traditional group (the TK group). The time of operation and the amount of bleeding during operation were recorded. Post-operative clinical outcomes and radiographic results were recorded and compared between these two groups. The Visual Analogue Scale (VAS), Neer score, Rating Scale of the American Shoulder and Elbow Surgeons, Constant-Murley score and complications such as infection, nonunion, implant loosening, fragment displacement and hardware pain were observed in the two groups.ResultsIn the comparison between the two groups, there was no significant difference in age, sex, the cause of displaced clavicle fracture, and other basic information between the two groups. The operation time, intraoperative fluoroscopy time, and intraoperative blood loss were significantly reduced in the NK group (P < 0.01). There were 2 cases of plate fracture in the TK group. The follow-up results showed that there was no significant difference in VAS, Neer score, ASES, and Constant-Murley scores between the two groups.ConclusionThe use of Nice knot, in comminuted and displaced clavicle fractures can reduce intraoperative blood loss, shorten operation time, facilitate intraoperative reduction, and achieve satisfactory postoperative clinical results. This study demonstrates that Nice knot is a simple, safe, practical and effective auxiliary reduction method.