Effects of HMGB1/TLR4 on secretion IL-10 and VEGF in human jaw bone-marrow mesenchymal stem cells

Abstract Objective: We aimed to investigate the regulatory effects of HMGB1/TLR4 signaling pathway on the expression of IL-10 and VEGF in human bone marrow mesenchymal stem cells. Methodology: Human JBMSCs were isolated and cultured. Then, HMGB1 was added into the JBMSCs culture medium, and the protein and mRNA expression levels of IL-10 and VEGF were assessed. Moreover, cells were pretreated with a specific TLR4 inhibitor (TAK-242), and the expression changes of IL-10 and VEGF were compared. Results: Compared with the control group, exposure to HMGB1 in human JBMSCs up-regulated TLR4, IL-10, and VEGF secretion at both protein and mRNA levels (P<0. 05). In addition, the increased expression of IL-10 and VEGF could be restrained in TAK-242 group compared with the HMGB1 group (P<0.05). Conclusions: The results indicated that HMGB1 activate TLR4 signaling pathway in Human JBMSCs, which plays a regulatory role in cytokines expression.

AF4/FMR2 and IL-10 gene single nucleotide polymorphisms are correlated with disease susceptibility and immune infiltration in ankylosing spondylitis / 南方医科大学学报

OBJECTIVE@#To explore the correlation of polymorphisms of AF4/FMR2 family genes and IL-10 gene with genetic susceptibility to ankylosing spondylitis (AS) and identify the high-risk factors of AS.@*METHODS@#This case-control study was conducted among 207 AS patients and 321 healthy individuals. The tag single nucleotide polymorphisms (SNPs) rs340630, rs241084, rs10865035, rs1698105, and rs1800896 of the AF4/FMR2 family gene and IL-10 gene of the AS patients were genotyped, and the distribution frequencies of the genotypes and alleles were analyzed to explore the relationship between different genetic models and AS and the gene-gene and gene-environment interactions.@*RESULTS@#Gender ratio, smoking history, drinking history, hypertension, erythrocyte sedimentation rate and C-reactive protein differed significantly between the case group and the control group (P < 0.05). The dominant model and recessive model of AFF1 rs340630, the recessive model of AFF3 rs10865035, and the recessive model of IL-10 rs1800896 were significantly different between the two groups (P=0.031, 0.010, 0.031, and 0.019, respectively). Gene-environment interaction analysis suggested that the interaction model incorporating AFF1 rs340630, AFF2 rs241084, AFF3 rs10865035, AFF4 rs1698105, IL-10 rs1800896, smoking history and drinking history was the best model. The genes related with AF4/FMR2 and IL-10 were enriched in the biological processes of AF4 super extension complex, interleukin family signal transduction, cytokine stimulation and apoptosis. The expression levels of AF4/FMR2 and IL-10 were positively correlated with immune infiltration (r > 0).@*CONCLUSION@#The SNPs of AF4/FMR2 and IL-10 genes are associated with the susceptibility to AS, and the interactions of AF4/FMR2 and IL-10 genes with the environmental factors contributes causes AS through immune infiltration.

Diagnosis Significance of the Levels of Cytokines IL-6, IL-10 and CXCL-13 in Cerebrospinal Fluid for Central Nervous System Infiltration of Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To evaluate the diagnostic value of the expression levels of cytokines interleukin-6(IL-6), interleukin-10 (IL-10) and chemokine （C-X-C motif） ligand-13 (CXCL-13) in cerebrospinal fluid (CSF) for central nervous system infiltration of lymphoma.@*METHODS@#Forty patients diagnosed as lymphoma or acute lymphoblastic leukemia in General Hospital of Northern Theater Command from July 2020 to July 2021 were collected and recorded their CSF indexes, including pressure, protein, Pandy test, nucleated cell count, glucose and chlorine content in CSF. The levels of cytokines IL-6, IL-10 and CXCL-13 were detected by Enzyme-linked immunosorbent assay.@*RESULTS@#The patients were divided into CNSI (central nervous system infiltration) group and non-CNSI group, the average levels of IL-6, IL-10, CXCL-13 and IL-10/IL-6 ratio in CNSI group were higher than those in non-CNS group, but the difference of IL-10/IL-6 ratio between the two groups was statistically significant (P<0.05). Then the patients were divided into protein elevated(n=14) group and protein normal group(n=26), the levels of IL-6 ［ (5.78±2.69) pg/ ml］ and CXCL-13 ［(0.83±0.59) pg/ml］ in protein elevated group were significantly higher than those in the protein normal group ［IL-6: (2.41±1.16) pg/ml; CXCL-13: (0.38±0.18) pg/ml］ (P<0.05). Further analysis of the expression levels of the cytokines in non-CNSI group (n=32), IL-6, IL-10, CXCL-13 level and IL-10/IL-6 ratio in the protein elevated group (n=12) were higher than those in the protein normal group (n=20), but the difference was not statistically significant.@*CONCLUSION@#The levels of IL-6, IL-10 and CXCL-13 in CSF of lymphoma patients with CNS infiltration were higher than those in non-CNS infiltration group, and those in patients with protein elevated group are higher than those in the protein normal group.

Relationship between IL-17, TNF-α, IL-10, IFN-γ, and IL-18 polymorphisms with the outcome of hepatitis B virus infection in the Turkish population

SUMMARY OBJECTIVE: Hepatitis B virus is a global threat that can lead to liver cirrhosis and hepatocellular carcinoma. For the treatment of chronic hepatitis B virus, polymorphisms might be an option for gene treatments. This study aimed to investigate the effects of IL-17, TNF-α, IL-10, IFN-γ, and IL-18 gene polymorphisms on hepatitis B virus infection in the Turkish population. METHODS: The genotypes and allele distribution of 75 patients exposed to hepatitis B virus and 50 healthy control individuals were analyzed. The real-time polymerase chain reaction method was used for identification. RESULTS: A correlation was observed between susceptibility to hepatitis B virus infection and IL-17 Exon 3/3'UTR (rs1974226) C, IL-17 Exon 3 (rs763780) A, IL-18 (-607) (rs1946518) A alleles, and IL-17 Exon 3 (rs763780) AA genotype (p=0.006, p=0.009, p=0.025, and p=0.008, respectively). Furthermore, IL-18 (-137) (rs187238) TT genotype and TNF-α-308 (rs1800629) G and A alleles, were associated with protection against hepatitis B virus infection (p=0.0351 and p=0.032, respectively). CONCLUSION: This study demonstrated that TNF-α (-308), IL-17 (Exon 3/3' UTR), IL-17 (Exon 3), and IL-18 (-607) polymorphisms are associated with hepatitis B virus infection. Therefore, these may serve as potential therapeutic targets for chronic viral hepatitis in the Turkish population.

Effect of Qiling Baitouweng Tang on Proliferation and Apoptosis in Diffuse Large B-cell Lymphoma Through JAK2/STAT3 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Qiling Baitouweng Tang （QLBTWT） on proliferation and apoptosis， Janus kinase 2 （JAK2）/signal transducer and activator of transcription 3 （STAT3） signaling pathway and interleukin-10 （IL-10） in diffuse large B-cell lymphoma （DLBCL）. MethodWith human DLBCL cells OCI-LY10 and U2932 as research objects， cell proliferation was detected by cell counting kit-8 （CCK-8） assay. After treatment with 0， 4.6， 9.3， 18.7， 37.5， 75， 150 mg·L-1 QLBTWT for 24 h， the half-inhibitory concentration （IC50） of OCL-LY10 and U2932 cells was calculated to be 9.33， 16.13 mg·L-1， respectively， based on which， 9.5， 19， 38 mg·L-1 QLBTWT were selected for subsequent experiments. After 0， 9.5， 19， 38 mg·L-1 QLBTWT treatment for 24 h， the zymogen activities of Caspase-3， Caspase-8 and Caspase-9 in OCI-LY10 and U2932 cells were detected using corresponding activity assay kits （colorimetric）， and the IL-10 expression was detected by enzyme-linked immuno sorbent assay （ELISA）. The apoptosis rate and cell cycle of OCI-LY10 and U2932 cells treated with different concentrations of QLBTWT for 24 h were detected by flow cytometry. The expressions of apoptosis-related proteins ［B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved poly adenosine diphosphate ribose polymerase （cleaved PARP）， cleaved Caspase-3］， JAK2， STAT3， phospho-JAK2 （p-JAK2）， phospho-STAT3 （p-STAT3） pathway proteins， and c-Myc protein in OCL-LY10 and U2932 cells after 24 h treatment with 0， 9.5， 19， 38 mg·L-1 QLBTWT were all tested by Western blot. ResultAfter QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h， cell proliferation was inhibited in each QLBTWT group compared with that in the control group （P<0.05， P<0.01）. The zymogens of Caspase-3， Caspase-8 and Caspase-9 were activated （P<0.01）， and there was an increase in cell apoptosis （P<0.05， P<0.01） and cell cycle arrest at Gap phase1 （G1） phase in 9.5， 19 and 38 mg·L-1 QLBTWT group （P<0.05， P<0.01）. After 9.5， 19 and 38 mg·L-1 QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h， the expressions of Bcl-2， p-JAK2 and p-STAT3 proteins were decreased （P<0.01）， and the expressions of Bax， cleaved PARP and cleaved Caspase-3 proteins were increased （P<0.01）， but no significant change was observed in the expressions of JAK2 and STAT3 proteins. Compared with the conditions in the control group， the expressions of c-Myc， p-JAK2， and p-STAT3 proteins were down-regulated in 19 mg·L-1 QLBTWT group and 19 mg·L-1 QLBTWT+10 μg·L-1 IL-10 group （P<0.05， P<0.01）， and up-regulated in 10 μg·L-1 IL-10 group （P<0.05， P<0.01）， while there was no difference in JAK2/STAT3 proteins. ConclusionQLBTWT can inhibit proliferation and induce apoptosis of human DLBCL cells OCI-LY10 and U2932， and the potential mechanism may be related to the regulation of JAK2/STAT3 signaling pathway.

Evaluation of the efficiency of ciprofloxacin against S. Typhi by altering the production of cytokines in acute typhoid fever in patients at Al-Diwaniyah Hospitals, Iraq

Aims@#Typhoid fever is a life-threatening disease in the developing world that claims >600,000 deaths per year. Its causative agent Salmonella Typhimurium (S. Typhi) can be treated with ciprofloxacin, an effective broad-spectrum antibiotic that enhances the natural host defenses. However, the emergence of resistant bacterial strains may be a warning alarm against the clinical use of this antibiotic. This study was aimed to investigate the efficiency of ciprofloxacin treatment (250 mg/mL) against S. Typhi by altering the production of serum cytokines IL-10, 1L-6 and TNF-α in acute typhoid fever patients in Diwanyah Hospitals.@*Methodology and results@#ELISA and Western Blot methods were used to investigate cytokine levels in patients and healthy controls sera. Our results showed that all cytokines’ levels before treatment with ciprofloxacin were significantly higher than the control (healthy group). However, treated patients with ciprofloxacin revealed a significantly reduced concentration of IL-10 and TNF-α compared to untreated control samples. However, the level of IL-6 was higher even with ciprofloxacin treatment.@*Conclusion, significance and impact of study@#The study concluded that ciprofloxacin (250 mg/mL) might significantly alter serum cytokines IL-6, IL-10 and TNF-α levels in acute typhoid fever patients. Therefore, further molecular studies are essential to understand the effect of ciprofloxacin on the production of cytokines.

Study on dynamic expression of IL-10 on scarring after implantation of glaucoma drainage material / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the dynamic expression characteristics of interleukin-10(IL-10)after implantation of glaucoma drainage material, and to reveal the role of IL-10 on scarring formation.METHODS:Totally 75 New Zealand white rabbits were randomly divided into three groups, which were implanted with different types of material-Polymethyl methacrylate coated Parylene C(PMMA group), silicone together with injection of Mitomycin C(MMC)(silicon-MMC group)and silicone(silicone group). Aqueous humor were collected at 1, 3d, 1, 2, 3, 4 and 8wk after operation and enzyme-linked immunosorbent assay(ELISA)were utilized to detect the expression of IL-10 in the aqueous humor. The connective tissue surrounding the material were collected at 1, 2, 3, 4 and 8wk postoperatively. Hematoxylin-eosin(HE)staining was applied to evaluate the proliferation of fibroblasts and the infiltration of inflammatory cells. The protein expression and mRNA of IL-10 in the connective tissue were detected by immunohistochemistry and real-time PCR.RESULTS:Compared with PMMA and silicon-MMC group, silicone group showed significantly increased proliferation of fibroblasts and infiltration of inflammatory cells according to the HE staining result. The result of ELISA showed the expression of IL-10 in the aqueous humor increased significantly at the early stage after surgery, and then decreased gradually,the highest appeared on the third day after operation,and in silicone group there was higher than the other two groups in the early stage postoperatively(1d-3wk)(all <i>P</i><0.05), and there was no significant difference in the late stages(4-8wk). The protein expression and mRNA of IL-10 in connective tissue were the highest in the first week after operation, decreased gradually at 2-3wk after operation, and increased again at 4-8wk after operation by immunohistochemistry and real-time PCR. And the expression was higher in silicone group than in the other two groups at each time point(all <i>P</i><0.05). Furthermore, there was a positive correlation between the expression of IL-10 protein and the proliferation of fibroblasts in the late stages(4-8wk).CONCLUSION: After implantation of glaucoma drainage material, the process of IL-10 increased first, then decreased gradually, and increased again 4wk later, thus IL-10 may be a potential target for inhibiting the scar formation.

Suppression of L-arginine-induced acute necrotizing pancreatitis in rats by metformin associated with the inhibition of myeloperoxidase and activation of interleukin-10 / Supresión de la pancreatitis necrotizante aguda inducida por L-arginina en ratas por metformina asociada con la inhibición de mieloperoxidasa y activación de interleucina-10

SUMMARY: Acute pancreatitis is a frequent life-threatening inflammatory disease of the pancreas characterized by severe abdominal pain that lasts for days to weeks. We sought to determine whether the antidiabetic and anti-inflammatory drug, metformin can substantially protect against acute pancreatitis in an animal model of L-arginine-induced acute pancreatitis, and whether this is associated with the augmentation of the anti-inflammatory cytokine interleukin-10 (IL-10) and inhibition of the enzyme that promotes tissue damage, myeloperoxidase (MPO). Rats were either injected with two doses of the amino acid L-arginine (2.5 gm/kg; i.p., at one-hour intervals) before being sacrificed after 48 hours (model group) or were pretreated with metformin (50 mg/kg) daily for two weeks prior to L- arginine injections and continued receiving metformin until the end of the experiment (protective group). Using microscopic examination of the pancreas and blood chemistry, we observed that L-arginine induced acute pancreatic injury. This is demonstrated by an enlarged pancreas with patchy areas of haemorrhage, vacuolated cytoplasm and pyknotic nuclei in the acini, disorganized lobular architecture with infiltration of inflammatory cells within the interlobular connective tissue (CT) septa, and the presence of congested blood vessels that were substantially ameliorated by metformin. Metformin also significantly (p<0.05) inhibited L-arginine-induced MPO, lactate dehydrogenase (LDH), and the inflammatory biomarker tumor necrosis factor alpha (TNF-α). Whereas, metformin significantly (p<0.05) increased IL-10 levels that were inhibited by pancreatitis induction. We further demonstrated a significant (p<0.001) correlation between the scoring of the degree of pancreatic lobules damage tissue damage and the blood levels of TNF-α, IL-10, LDH, and MPO. Thus, metformin effectively protects against L-arginine-induced acute pancreatitis, which is associated with the inhibition of MPO and augmentation of IL-10.

RESUMEN: La pancreatitis aguda es una enfermedad inflamatoria del páncreas que amenaza la vida y se caracteriza por un dolor abdominal intenso que dura de días a semanas. Buscamos determinar si la metformina, fármaco antidiabético y antiinflamatorio, puede proteger contra la pancreatitis aguda en un modelo animal de pancreatitis aguda inducida por L-arginina. Además se estudió la asociación con el aumento de la citocina antiinflamatoria interleucina-10. (IL-10) e inhibición de la enzima que promueve el daño tisular, mieloperoxidasa (MPO). Las ratas se inyectaron con dos dosis del aminoácido L-arginina (2,5 g / kg; ip, a intervalos de una hora) antes de ser sacrificadas des- pués de 48 horas (grupo modelo) o se pre trataron con metformina (50 mg / kg) durante dos semanas antes del tratamiento de L- arginina y continuaron recibiendo metformina hasta el final del experimento (grupo protector). Mediante el examen microscópico del páncreas y la química sanguínea, se observó que la L- arginina inducía una lesión pancreática aguda. Se observó un aumento significativo de tamaño del páncreas con áreas hemorrágicas, citoplasma vacuolado y núcleos picnóticos en los acinos, arquitectura desorganizada con infiltración de células inflamatorias dentro de los tabiques del tejido conjuntivo interlobulillar (TC) y la presencia de vasos sanguíneos congestionados mejorados por metformina. Se observó que la metformina inhibió significativamente (p <0,05) la MPO inducida por L- arginina, la lactato deshidrogenasa (LDH) y el factor de necrosis tumoral alfa (TNF-α). Además, demostramos una correlación significativa (p <0,001) entre la puntuación del grado de daño tisular de los lóbulos pancreáticos y los niveles sanguíneos de TNF-α, IL-10, LDH y MPO. Por tanto, la metformina protege eficazmente contra la pancreatitis aguda inducida por L-arginina, que se asocia con la inhibición de MPO y el aumento de IL-10.

Expansion of T regulatory lymphocytes by murine bone marrow dendritic cells previously stimulated with Anisakis simplex larval antigens

BACKGROUND Anisakis simplex antigens present immunomodulatory properties by the induction of tolerogenic dendritic cells (DCs) in mice. OBJECTIVES To study the capacity of DCs stimulated with A. simplex excretory-secretory (ES) or crude extract (CE) to generate Tregs. To investigate in vitro effects of antigens on the metabolic activity of splenocytes induced by LPS or CpG. METHODS Phenotypic and functional characterization of T cells co-cultured with A. simplex-pulsed DCs was performed by flow cytometry. Lymphocyte mitochondrial respiratory activity was estimated by the Alamar Blue® Assay. FINDINGS In C57BL/6J, CD4+CD25-Foxp3+ and CD8+CD25-Foxp3+ populations increased by CE-stimulated-DCs. In BALB/c, CE-stimulated-DCs caused the expansion of CD4+CD25+Foxp3+IL-10+ and CD8+CD25+Foxp3+IL-10+. IFN-γ expression raised in BALB/c CD4+CD25+ and CD4+CD25- for CE and ES, respectively. ES-stimulated-DCs increased CD4+CD25+ Foxp3+ and CD8+CD25- Foxp3+ expression in T cells. The association of ES or CE with LPS produced the increase in splenocyte activity in C57BL/6J. The association of CE with CpG decreased the proliferation caused by CpG in C57BL/6J. MAIN CONCLUSIONS A. simplex increase the frequency of Tregs, which in turn produce IL-10 and IFN-γ. The host genetic base is essential in the development of anti-Anisakis immune responses (Th2, Th1, Treg).

Impact of Allium sativum ethanol extract on immuno-regulatory T cells and anti-inflammatory cytokine profile in murine schistosomiasis

@#Parasite immune response against schistosomal antigens involves both the innate and adaptive immune response. Tregs have a suppressive effect and play a role on the parasite’s immune evasion. This study aimed to evaluate active compounds of Allium sativum (AS) ethanol extract and the impact of AS extract alone or in combination with praziquantel on Tregs and anti-inflammatory cytokines TGF-β and IL-10 in mice infected with S. mansoni. Phytochemical screening of AS bulbs for various active constituents and qualitative and quantitative analysis of the flavonoids and phenolic acids were done using HPLC. Measurement of splenocytes Treg cell phenotypes and anti-inflammatory cytokines TGF-β and IL-10 was done by flow cytometric analysis. The data are expressed as mean ± SD. Statistical significance was determined by one-way ANOVA utilizing the statistical package (SPSS version 17.0). HPLC of AS ethanol extract revealed presence of 22 and 18 compounds of flavonoids and phenolic acids, respectively. S. mansoni infection upregulated the Treg cells subsets (CD4, CD25, Foxp3) frequencies and the levels of TGF-β and IL-10 anti-inflammatory cytokines when compared to healthy control. AS ethanol extract alone or combined with PZQ decreases the production of Treg cells from spleen in addition to the reduction in antiinflammatory cytokines IL-10 and TGF-β. This study recommends that the combination of AS ethanol extract and PZQ may play a role in maintaining the homeostasis of the immune system during schistosomiasis by decreasing Treg cells and anti-inflammatory cytokines IL10 and TGF-β production.

Effect of bone marrow mesenchymal stem cell on the expression of IL-10 and TNF-α in mice with ischemia-reperfusion acute kidney injury / 器官移植

Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.

Changes and clinical significance of four biomarkers in gingival crevicular fluid after nonsurgical periodontal therapy / 口腔疾病防治

Objective@# To study the changes in levels of interleukin (IL)-6, IL-10, tumor necrosis factor-alpha (TNF-α), and alkaline phosphatase (ALP) in the gingival crevicular fluid (GCF) of patients with severe chronic periodontitis before and after nonsurgical periodontal therapy and to explore the relationship among the levels of these four biomarkers in GCF, their periodontal status and their clinical significance to evaluate the effect of nonsurgical periodontal therapy and periodontitis activity.@*Methods@# In total, 30 patients with severe chronic periodontitis were enrolled in a 1-year longitudinal pilot study (Chinese Clinical Trial Registry: ChiCTR-OCH-13004679). At baseline and 1, 3, 6, and 12 months after nonsurgical therapy, the periodontal clinical indicators plaque index (PLI), probing depth (PD), clinical attachment loss (CAL), sulcus bleeding index (SBI) were recorded. Filter paper strips were used to collect two deep-pocket (probing depth ≥ 6 mm) and two shallow-pocket (probing depth ≤ 4 mm) periodontal sites for each patient and weighed. The levels of interleukin IL-6, IL-10, TNF-α, and ALP in GCF were assessed using enzyme-linked immunosorbent assay. Meanwhile, 30 healthy sites of 15 subjects with healthy periodontium were used as the baseline controls for patients with severe chronic periodontitis.@*Results @#At the baseline, the TNF-α, ALP and IL-6 levels in GCF of the disease sites of patients with periodontitis were significantly higher than those in healthy periodontal sites of the control group (P < 0.001), and the levels of IL-10 were significantly lower than those in the control group (P < 0.001). In patients with severe chronic periodontitis, the levels of TNF-α, ALP and IL-6 in GCF at deep-pocket sites were significantly higher than those at shallow-pocket sites (P <0.001), and the IL-10 levels were significantly lower than those at shallow-pocket sites (P < 0.001). 1, 3, 6, and 12 months after nonsurgical treatment, the levels of TNF-α and ALP in GCF at the shallow- and deep-pocket sites in patients with chronic periodontitis significantly decreased, the level of IL-10 significantly increased (P < 0.005), and the level of IL-6 in GCF at the deep-pocket sites significantly decreased (P < 0.005). However, there was no significant difference in IL-6 level at shallow-pocket sites (P > 0.05). 1, 3, 6, and 12 months after nonsurgical treatment, the periodontal clinical indicators were improved compared with the baseline. In addition, there was a significant correlation between the levels of these four biomarkers and the periodontal clinical parameters (P < 0.05). During the two follow-up visits after nonsurgical periodontal therapy, the sites with more than 2-mm increase in attachment loss had significant differences in the levels of the four biomarkers in the GCF compared with the previous visit time (P < 0.005).@*Conclusion@#The detection of the levels of these four biomarkers in GCF has strong clinical significance for assessing the severity of periodontitis and the efficacy of nonsurgical periodontal therapy. Increased levels of TNF-α, ALP, and IL-6 and decreased IL-10 levels in GCF may indicate periodontitis progression at this site.

Activation of death receptor 3 signaling pathway prevents antibody-mediated transfusion-related acute lung injury via αDR3 antibody / 中国输血杂志

【Objective】 To explore the efficacy and possible mechanisms of activation of Death receptor 3 (DR3) signaling pathway in the prevention of antibody-mediated transfusion-related acute lung injury (TRALI) via DR3 agonistic (αDR3) antibody. 【Methods】 8-10-week-old male wild-type Balb/c mice (40) were randomly divided into Naïve group, isotype control group, TRALI model group, and intervention group. Mice without any treatment served as Naïve group. Isotype and TRALI model were established by intraperitoneally priming 8-10-week Balb/c mice with LPS 18 h prior to injection of an IgG2a isotype antibody and anti-MHC-Ⅰ antibody via tail vein, respectively. Intervention group: mice were intraperitoneally injected with a single dose of αDR3 antibody (1 mg/kg) on day 1; after 3 days, the mice were challenged with LPS 18 h prior to injection of an anti-MHC-I antibody. The lung tissues and spleens of mice in each group were collected at the mice died or 2 hours after TRALI modeling for the lung injury severity. Spleens were collected to measure the proportion of Treg by flow cytometry. Foxp3, iNOS, and CD206 immunohistochemical staining combining with optical density analysis of lung tissues were used to represent Treg, M1 macrophages and M2 macrophages, respectively. The concentration of IL-6, IL-1β, TNF-α, and IL-10 cytokines in lung tissues was detected via Cytometric Beads Array. 【Results】 Compared with TRALI group, 1) the lung injury of mice were significantly alleviated in intervention group; 2) the proportion of Treg(%) in the spleens (9.295±1.349 vs 2.257±0.610, P<0.05), Foxp3 expression of Treg in the lungs (0.302 6±0.052 6 vs 0.230 2±0.016 3, P<0.05), and the concentration of Treg derived cytokines IL-10 in the lungs (29.52±8.885 vs 8.045±1.911, P<0.05) increased significantly in intervention group; 3) the iNOS expression of M1 macrophages (0.209 6±0.013 9 vs 0.279 6±0.045 2) and the concentration of M1 macrophage derived cytokines IL-6 (23.22±19.35 vs 301.1±157.7), IL-1β (46.76±25.34 vs 307.6±183.8), and TNF-α (45.99±14.16 vs 143.9±44.43) in the lungs was significantly reduced(P<0.05), while CD206 expression of M2 macrophages (0.291 2±0.032 1 vs 0.221 5±0.012 7) and the concentration of M2 macrophage derived IL-10 cytokines (29.52±8.885 vs 8.045±1.911) in the lungs increased significantly in intervention group(P<0.05). 【Conclusion】 Activation of DR3 signaling pathway by αDR3 antibody prevents antibody-mediated TRALI via expanding Treg, which regulates macrophage polarization by IL-10 derived from Treg.

Regulatory lymphocytes: the dice that resolve the tumor endgame

A large number of cancer patients relapse after chemotherapeutic treatment. The immune system is capable of identifying and destroying cancer cells, so recent studies have highlighted the growing importance of using combinatorial chemotherapy and immunotherapy. However, many patients have innate or acquired resistance to immunotherapies. Long-term follow-up in a pooled meta-analysis exhibited long-term survival in approximately 20% of patients treated with immune checkpoint inhibitors or the adoptive transfer of chimeric T cells. It has been reported that high levels of immunoregulatory cells in cancer patients contribute to immunotherapy resistance via immunosuppression. Among the most important regulatory cell subtypes are the CD4+ T-regulatory cells (Tregs), identified by their expression of the well-characterized, lineage-specific transcription factor FOXP3. In addition to CD4+ Tregs, other regulatory cells present in the tumor microenvironment, namely CD8+ Tregs and IL10-producing B-regulatory cells (Bregs) that also modulate the immune response in solid and lymphoid tumors. These cells together have detrimental effects on tumor immune surveillance and anti-tumor immunity. Therefore, targeting these regulatory lymphocytes will be crucial in improving treatment outcomes for immunotherapy.

Evaluation of Some Cytokines, CD4, Hepcidin, Iron Profile and Some Haematological Parameters of Pulmonary Tuberculosis Patients Coinfected with HIV in Southeast of Nigeria

The study was done to determine the levels of interferon-gamma, interleukin 6, interleukin 10, iron status, hepcidin and haematologicalparameters of patients with pulmonary tuberculosis co-infected with human immunodeficiency virus in Southeast, Nigeria. This study was carried out at the directly observed treatment-short course Tuberculosis (TB DOTS) centre of Federal Medical Centre, Umuahia, located in South-Eastern Nigeria. Therefore, sample size of 240 was used to give room for attrition. A total of two hundred and forty (240) subjects aged 18-60 years were enlisted for this study. Seven milliliters (7ml) of venous blood was collected from each subject and 2.5ml was dispensed into bottles containing di-potassium salt of ethylenediamine tetra-acetic acid (K2-EDTA) and was used for full blood count, CD4 count and HIV screening. Also, 4.5ml was dispensed into plain tubes. Serum was obtained after clotting by spinning at 3000 RPM for 10 minutes and was used for interferon gamma, interleukin-6, and interleukin-10, iron and hepcidin determination. Data was analysed using statistical package for social science (SPSS) version 20. Student t-test, ANOVA (Analysis of Variance), Pearson Product Moment and Chi-Square were the tools employed. Results were expressed as mean ± standard deviation and are presented in table and significance level was set at P<0.05.The results showed difference that was statistically significant (P<0.05) in IFN-γ (P=0.000), IL-6 (P=0.000) IL-10 (P=0.000), CD4 (P=0.000), hepcidin (P=0.000), Iron (P=0.000), TIBC (P=0.000), %TSA (P=0.001) ,WBC (P=0.000), Neutrophils (P=0.000), Lymphocyes (P=0.000), Monocytes (P=0.000), Eosinophils (P=0.000), Basophils (P=0.018), RBC (P=0.000), haemoglobin (P=0.000), PCV (P=0.000), MCV (P=0.000), MCH (P=0.000), MCHC (P=0.000), Platelets (P=0.000), ESR (P=0.000) when compared among control, TB, HIV and TB-HIV subjects respectively. The co infection of HIV on pulmonary TB patients increases the levels of the cytokines. The cytokines and hepcidin can be used as adjunct to prognostic and diagnostic markers as their levels decreased with increased duration of treatment of the patients. The study hasshown wide variations in the haemtological indices studied

Role of biomarkers in predicting anastomotic leakage following colorectal surgeries

Background: Recovery after surgery for patients with colorectal disease has improved with the advent of minimal access surgery and standardized recovery protocols. Despite these advances, anastomotic leakage remains one of the most dreaded complications following colorectal surgery, with rates of 3-27 per cent depending on specific risk factors. The aim of the study was to assess sensitivity and specificity of systemic and peritoneal drain-fluid bio-markers in early prediction of anastomotic leak; and to co-relate rise in levels of biomarkers and severity of clinical symptoms in patients who have undergone colo-rectal surgeries.Methods: The present study was a prospective observational study conducted on 60 patients in the Postgraduate Department of Surgery, Government Medical College, Srinagar after obtaining due ethical clearance over a period of two years.Results: The mean age was 54.87±11.901 years with 44 patients (73.3%) were males. Among systemic makers: the mean CRP level was 2.7800±0.500 mg/L, the mean total leukocyte count was 10.783±0.940 thousands and the mean serum procalcitonin level was 0.365±0.1385 ng/ml. Among peritoneal fluid drain bio-makers, the mean IL-6 level was 3551.066±1311.965 pg/ml, the mean IL-10 level was 628.533±460.358 pg/ml and the mean TNF-a level was 16.391±6.736 pg/ml. The anastomotic leak after colo-rectal surgery was noted in 16 patients (26.7%). In our study significant co-relation was noted between the rise in levels of peritoneal drain fluid biomarkers and severity of clinical symptoms but no significant co-relation was noted between the rise in levels of systemic markers and severity of clinical symptoms in patients who have undergone colo-rectal surgeries.Conclusions: Systemic biomarkers are poor predictors of anastomotic leak after colorectal surgery. But sensitivity and specificity of peritoneal fluid drain biomarkers in predicting anastomotic leak was significantly high.

Investigation of ADAMTS-1, BAX and IL-10 expressions in granulosa cells determinanting fertilization success in IVF / Investigación de las expresiones ADAMTS-1, BAX e IL-10 en célula de la granulosa determinando el éxito de fertilización en FIV

Granulosa cells (GCs) are essential components of follicles and play a role in regulating follicle development. The aim of this study was to investigate certain cellular components involved in the proliferation, differentiation and functional characteristics of granulosa cells in the success of fertilization of human oocytes during invitro fertilization (IVF) via immunohistochemical techniques. In this study, 30 patients who were diagnosed as primary or secondary infertility, polycystic ovary syndrome in the IVF center of Memorial Hospital, Department of Obstetrics and Gynecology were included. The amount of Anti Müllerian Hormone (AMH) in blood and granulosa cell diameter and cell core diameter were measured in 20 cells collected from each patient. In addition, degeneration scoring and BAX, ADAMTS-1, IL-10 expressions in granulosa cells were evaluated (p <0.01). It was thought that apoptosis induced by human GCs might be an indicator of egg quality. Moderate expression of ADAMTS-1 was thought to be related to failure of ovulation, deterioration of oocyte quality and decreased fertilization rate. This decrease in AMH levels may be associated with defects in granulosa cells. Therefore, significantly lower AMH secretion and increase in IL10 expression levels in healthy people can be explained by the increase of granulocyte cells.

Las células de la granulosa (GC) son componentes esenciales de los folículos y tienen un papel en la regulación del desarrollo de éste. El objetivo del estudio fue investigar ciertos componentes celulares involucrados en la proliferación, diferenciación y características funcionales de las células de la granulosa en el éxito de la fertilización de los ovocitos humanos durante la fertilización in vitro (FIV) a través de técnicas inmunohistoquímicas. En este estudio, se incluyeron 30 pacientes diagnosticados con infertilidad primaria o secundaria, síndrome de ovario poliquístico en el centro de FIV del Departamento de Obstetricia y Ginecología del Hospital Memorial. La cantidad de Hormona Anti Mülleriana (AMH) en la sangre, el diámetro de las células de la granulosa y el diámetro del núcleo celular se midieron en 20 células obtenidas de cada paciente. Además, se evaluó la puntuación de degeneración y las expresiones BAX, ADAMTS-1, IL-10 en células de granulosa (p <0,01). Se estimó que la apoptosis inducida por los GC humanos podría ser un indicador de la calidad del huevo. Se estimó que la expresión moderada de ADAMTS-1 estaba relacionada con el fracaso de la ovulación, el deterioro de la calidad de los ovocitos y la disminución de la tasa de fertilización. La disminución en los niveles de AMH puede estar asociada con defectos en las células de la granulosa. Por lo tanto, el aumento de las células de granulocitos puede explicar la disminución significativa de la secreción de AMH y el aumento de los niveles de expresión de IL10 en personas sanas.

Evaluating the effects of vanadyl sulfate on biomarkers of oxidative stress and inflammation in renal tissue of rats with diabetes type 2

Vanadyl sulfate (VS) is an ingredient in some food supplements and experimental drugs. This study was designed to assay the effects of VS on biomarkers of oxidative stress and inflammation in renal tissue of rats with diabetes type 2. 30 male Wistar rats were divided into three equal groups as follow: non-diabetics, non-treated diabetics and VS-treated diabetics. Diabetes type 2 has been induced through high fat diet and fructose in the animals. Diabetic rats were treated with 25 mg/kgBW of VS in water for 12 weeks. At the end of study, glucose and insulin were measured using commercially available kits in serum and biomarkers of oxidative stress and inflammation in renal homogenates of animals were measured by related methods. Compared to controls, glucose and insulin were increased significantly in non-treated diabetic rats (p-value <0.05) that showed the induction of diabetes type 2 in rats. The results showed that in VS-treated diabetic rats compared to the non-treated diabetic group, vanadyl sulfate significantly reduced the glucose and insulin secretion and changed renal inflammatory and oxidative markers, except protein carbonyl so that we couldn't find any significant changes. Our study showed that vanadyl supplementation had positive effects on oxidative stress and inflammation biomarkers in kidney of diabetic rats

Upregulation of IL-10 expression inhibits the proliferation of human periodontal ligament stem cells

Abstract: The abnormal increase in proliferation rate of human periodontal ligament stem cells (PDLSCs) is considered to be involved in the pathogenesis of periodontitis, a disease in which the IL-10-mediated anti-inflammatory pathway plays a critical role. This study aimed to investigate the involvement of microRNA-466l in periodontitis and to explore the possible interaction between IL-10 and microRNA-466l. PDLSCs were obtained from periodontitis-affected teeth and healthy control teeth. The expression of microRNA-466l and IL-10 mRNA was measured in PDLSCs using RT-qPCR. The proliferation ability of PDLSCs was analyzed using CCK-8 assays. Overexpression of microRNA-466l in a PDLSC cell line was established using two different types of PDLSCs, and the effect of microRNA-466l overexpression on IL-10 expression and cell proliferation were detected by western blot and CCK-8 assays, respectively. We found that expression levels of microRNA-466l and IL-10 mRNA were significantly lower (P < 0.05) in PDLSCs derived from periodontitis-affected teeth compared to those derived from healthy teeth. However, the cell proliferation ability was significantly higher in the PDLSCs derived from periodontitis-affected teeth. Meanwhile microRNA-466l overexpression decreased cell proliferation rates of both types of PDLSCs and upregulated IL-10 expression. Together, these data suggest that microRNA-466l can upregulate IL-10 and reduce the proliferation rate of PDLSCs.