Toll-like receptor 2-mediated downstream cytokine levels as determinant of malaria pathogenesis

Background & objectives: Toll-like receptors (TLRs) are transmembrane proteins that recognize specific molecular patterns and activate downstream cytokine production usually for the eradication of invading pathogens. The objective of this study was to evaluate the genetic polymorphism of TLR2 Arg753Gln (rs 5743708) and soluble cytokines and TLR2 expression levels in malaria disease cases. Methods: The study included prospectively collected 2 ml blood samples from 153 individuals clinically suspected for malaria and confirmed by microscopy and RDT from Assam. Stratification of the study groups was done as healthy control (HC, n=150), uncomplicated malaria (UC-M, n=128) and severe malaria (SM, n=25). The PCR-restriction fragment length polymorphism (RFLP) method was applied for the analysis of TLR2 Arg753Gln polymorphism and following the ELISA for soluble serum TLR2 (sTLR2) and its associated downstream cytokines, viz. tumour necrosis factor (TNF)-? and interferon (IFN)-? levels. Results: Variation in TLR2 Arg753Gln gene showed no association with the susceptibility and the severity of malarial infection. Soluble TLR2 expression was significantly higher in uncomplicated malaria (UC-M) cases compared to healthy controls (P=0.045) and in terms of SM cases, the expression was also found to be higher in UC-M cases (P=0.078). The TNF-? expression was significantly higher in SM cases compared to both UC-M and control (P=0.003 and P=0.004). Similarly, significantly elevated expression of IFN-? was noted in SM cases compared to both UC-M (P=0.001) and healthy controls (P<0.001). Interpretation & conclusions: The present study suggests the association of deregulated TLR2 pathway that leads to the deleterious downstream immune response in the development of malarial pathogenicity.

Interferon-gamma gene diplotype (AA-rs2069716 / AG-rs2069727) may play an important role during secondary outcomes of severe dengue in Brazilian patients

ABSTRACT Dengue is a global and growing health threat, especially in Southeast Asia, West Pacific and South America. Infection by the dengue virus (DENV) results in dengue fever, which can evolve to severe forms. Cytokines, especially interferons, are involved in the immunopathogenesis of dengue fever, and so may influence the disease outcomes. The aim of this study was to investigate the association between severe forms of dengue and two single nucleotide polymorphisms (SNPs) in the interferon-gamma gene (IFNG): A256G (rs2069716) and A325G (rs2069727). We included 274 patients infected with DENV serotype 3: 119 cases of dengue without warning signs (DWoWS), and 155 with warning signs (DWWS) or severe dengue (SD). DNA was extracted, and genotyped with Illumina Genotyping Kit or real time PCR (TaqMan probes). We estimated the adjusted Odds Ratios (OR) by multivariate logistic regression models. When comparing with the ancestral AA/AA diplotype (A256G/A325G), we found a protective association of the AA/AG against DWWS/SD among patients with secondary dengue (OR 0.51; 95% IC 0.24-1.10, p = 0.085), adjusting for age and sex. The variant genotype at locus A325G of the IFNG, in combination with the ancestral genotype at locus A256G, can protect against severe clinical forms of secondary dengue in Brazilian DENV3-infected patients.

The clinical utility of tuberculin skin tests: a single-center experience

Background: Tuberculin skin test (TST) has played an essential in the diagnosis of latent tuberculosis infection (LTBI) for nearly a century. Objective: This study aimed to investigate the general characteristics of patients tested with TST in a tertiary hospital within two years. Methods: All patients who were evaluated to screen for tuberculosis and received a TST were included. The Mantoux method was used for TST administration. Results: A total of 661 patients, 345 (52.2%) men and 316 (47.8%) women, with a mean age of 43.0 ±15.9 years, were included in the study. Accordingly, TST was performed prior to anti-TNF biological agent therapy for 50% (331) of the participants, for LTBI screening before solid organ and/or hematological stem cell transplantation for 20.4% (135), for screening following contact with tuberculosis for 25.1% (166), for screening of healthcare professionals for 1.1% (7), and medical report for 3.3% (22). 2.7% of the patients who took TST were diagnosed with active tuberculosis (14 with pulmonary tuberculosis and 4 with extrapulmonary tuberculosis). QuantiFERON-TB Gold (QFT) test was performed in 332 (50.2%) patients with anergic TST results. According to TST and QFT test results, 28.3% (187) of the patients were started on tuberculosis prophylaxis. Conclusion: While TST is most performed for LTBI screening prior to biological agent therapy, almost one-fourth of patients taking TST require tuberculosis prophylaxis. On the other hand, about half of the patients require an additional QFT test.

Antecedentes: La prueba de la tuberculina ha jugado un papel fundamental en el diagnóstico de la infección latente por tuberculosis durante casi un siglo. Objetivo: Investigar las características generales de los pacientes a los que se les realizó la prueba de tuberculina en un hospital de tercer nivel. Métodos: Se incluyeron todos los pacientes que fueron incluidos en un tamizaje de tuberculosis mediante la prueba de tuberculina. Se utilizó el método de Mantoux para la administración de esta prueba. Resultados: Se incluyeron en el estudio un total de 661 pacientes, 345 (52.2%) hombres y 316 (47.8%) mujeres, con una edad media de 43.0 ±15.9 años. La prueba de tuberculina se realizó en el 50% (331) de los participantes, antes de la terapia con agentes biológicos anti-TNF; En el 20.4% (135) se hizo la prueba antes del trasplante de órganos sólidos y/o células madre hematológicas; para el 25.1% (166) se realizó tras contacto con la tuberculosis, el 1.1% (7) para tamizaje de los profesionales sanitarios y con informe médico para el 3.3% (22). El 2.7% de los pacientes que se realizaron la prueba de tuberculina fueron diagnosticados con tuberculosis activa (14 pulmonar y 4 extrapulmonar). La prueba QuantiFERON-TB Gold (QFT) se realizó en 332 (50.2 %) pacientes con resultados anérgicos para tuberculina. Según los resultados de las pruebas de tuberculina y QFT, el 28.3% (187) de los pacientes iniciaron profilaxis antituberculosa. Conclusión: Si bien la prueba de tuberculina se realiza comúnmente para la detección de tuberculosis latente antes de la terapia con agentes biológicos, casi una cuarta parte de los pacientes que se les hizo la prueba de tuberculina requieren profilaxis para tuberculosis. Por otro lado, aproximadamente la mitad de los pacientes requieren una prueba QFT adicional.

Potential anticancer peptides design from the cysteine rich plant defensins: An in silico approach-Supplementary Data

Cancer is the second leading cause of mortality worldwide preceded by cardiovascular diseases. The therapeutic approaches for drug developmentinclude the use of small molecules, antibodies, peptidesor short nucleic acid sequences. The peptide-based drugs have been developed to treat many diseases like cardiovascular diseases, cancer, metabolic disorders, immunological diseases and viral infections. More than 80 peptide drugs are already in the market. These therapeutic peptides have several important benefits over antibodies and proteins due to their small size, ease for chemical synthesis and further the ability to penetrate cell membrane. Furthermore, peptide drugs have high specificity, activity, and affinity. The plant defensins BcDef1, TPP3, NaD1, 2N2R and 2LR3 have been studied for their role in wide range of diseases. This study focussed on the conformation of plant defensins rich in disulfide bonds. The structure for BcDef1 has been predicted from the conformational ensemble. Then, we designed anticancer peptides from these defensins with computational methods. The designed anticancer peptides have been studied for their immunogenicity as well as homology with human proteome. The role of designed peptides has been suggested for interferon-gamma induction, the later has been shown to possess a very important role in cancer.

Potential anticancer peptides design from the cysteine rich plant defensins: An in silico approach-Supplementary Data

Cancer is the second leading cause of mortality worldwide preceded by cardiovascular diseases. The therapeutic approaches for drug developmentinclude the use of small molecules, antibodies, peptidesor short nucleic acid sequences. The peptide-based drugs have been developed to treat many diseases like cardiovascular diseases, cancer, metabolic disorders, immunological diseases and viral infections. More than 80 peptide drugs are already in the market. These therapeutic peptides have several important benefits over antibodies and proteins due to their small size, ease for chemical synthesis and further the ability to penetrate cell membrane. Furthermore, peptide drugs have high specificity, activity, and affinity. The plant defensins BcDef1, TPP3, NaD1, 2N2R and 2LR3 have been studied for their role in wide range of diseases. This study focussed on the conformation of plant defensins rich in disulfide bonds. The structure for BcDef1 has been predicted from the conformational ensemble. Then, we designed anticancer peptides from these defensins with computational methods. The designed anticancer peptides have been studied for their immunogenicity as well as homology with human proteome. The role of designed peptides has been suggested for interferon-gamma induction, the later has been shown to possess a very important role in cancer.

Potential anticancer peptides design from the cysteine rich plant defensins: An in silico approach

Cancer is the second leading cause of mortality worldwide preceded by cardiovascular diseases. The therapeutic approaches for drug developmentinclude the use of small molecules, antibodies, peptidesor short nucleic acid sequences. The peptide-based drugs have been developed to treat many diseases like cardiovascular diseases, cancer, metabolic disorders, immunological diseases and viral infections. More than 80 peptide drugs are already in the market. These therapeutic peptides have several important benefits over antibodies and proteins due to their small size, ease for chemical synthesis and further the ability to penetrate cell membrane. Furthermore, peptide drugs have high specificity, activity, and affinity. The plant defensins BcDef1, TPP3, NaD1, 2N2R and 2LR3 have been studied for their role in wide range of diseases. This study focussed on the conformation of plant defensins rich in disulfide bonds. The structure for BcDef1 has been predicted from the conformational ensemble. Then, we designed anticancer peptides from these defensins with computational methods. The designed anticancer peptides have been studied for their immunogenicity as well as homology with human proteome. The role of designed peptides has been suggested for interferon-gamma induction, the later has been shown to possess a very important role in cancer.

Pro-and Anti-inflammatory Biomarkers as Potential Prognosis Markers in Oral Squamous Cell Carcinoma

ABSTRACT: The most widely used method to classify prognostic factors in cancers today is TNM. However, Oral Squamous Cell Carcinoma (OSCC) often demonstrates different behaviors in relation to aggressiveness and therapeutic response at the same TNM stage. So, in such cases biomarkers can be used to identify the biological diversity of these tumors more reliably, leading to better therapeutic strategies and disease management. The presence of inflammatory immune cells in the tumor microenvironment can have pro or antitumor effects and the investigation of the expression of inflammatory markers in OSSC can be usefulto design immunotherapeutic interventions. The Transforming Growth Factor alpha is a potent stimulator of cell migration that acts on cell proliferation, invasion and metastasis of cancer, as well as immune suppression and angiogenesis. Inflammatory cytokines, such as Interferon-gamma, mediate macrophage differentiation. Macrophages are an important component of the OSCC microenvironment. The greater amount of tumor-associated macrophages, especially the M2 phenotype, may be associated with a more aggressive biological behavior of the OSCC and, consequently, with reduced survival.

RESUMEN: El método más utilizado para clasificar los factores de pronóstico en los cánceres en la actualidad es TNM. Sin embargo, el carcinoma oral de células escamosas (COCE) a menudo muestra diferentes comportamientos en relación con la agresividad y la respuesta terapéutica en la misma etapa TNM. Entonces, en tales casos, los biomarcadores pueden usarse para identificar la diversidad biológica de estos tumores de manera más confiable, lo que lleva a mejores estrategias terapéuticas y manejo de la enfermedad. La presencia de células inmunes inflamatorias en el microambiente tumoral puede tener efectos pro o antitumorales y la investigación de la expresión de marcadores inflamatorios en COCE puede ser útil para diseñar intervenciones inmunoterapéuticas. El factor de crecimiento transformante α es un potente estimulador de la migración celular que actúa sobre la proliferación celular, la invasión y metástasis del cáncer, así como la inmunosupresión y la angiogénesis. Las citocinas inflamatorias, como el IFN-γ, median en la diferenciación de macrófagos. Los macrófagos son un componente importante del microambiente COCE. La mayor cantidad de macrófagos asociados a tumores, especialmente el fenotipo M2, puede estar asociada a un comportamiento biológico más agresivo del COCE y, en consecuencia, a una menor supervivencia.

Changes of humoral and cellular immune indexes in peripheral blood of children with Kawasaki disease at different stages / 中国医师进修杂志

Objective:To investigate the dynamic change characteristics of peripheral blood interferon-γ (INF-γ), interleukin (IL)-4 levels and T helper cell (Th)1/Th2 balance in acute, subacute and restoration stages of children with Kawasaki disease (KD).Methods:Forty-one children with KD received treatment in Women′s and Children′s Hospital Affiliated of School of Medicine University of Electronic Science and Technology of China, Chengdu Women′s and Children′s Central Hospital from May 2017 to January 2020 were enrolled as the observation group, and 41 healthy children examinee from the same period were enrolled as the control group. Children with KD of the observation group were performed with tuberculin pure protein derivative (PPD) test when acute and restoration stage of KD respectively. Peripheral venous blood were drawn from KD children of the observation group in acute, subacute and restoration stage and the control group respectively, serum immune globulin IgG, IgA, IgM and IgE, serum IFN-γ and IL-4 levels were detected by enzyme linked immunosorbent assay (ELISA).Results:Positive rates of PPD test in the restoration stage was higher than that in the acute stage: 65.85%(27/41) vs.17.07%(7/41), there was statistical difference ( χ2 = 20.10, P<0.05). The levels of serum IgG, IgA, IgM and IgE in the acute stage , subacute stage and restoration stagewere gradually decreased ( P<0.05). The levels of serum IgG, IgA, IgM and IgE in the restoration stage and the control group had no significant differences ( P>0.05). The levels of serum IFN-γ and IFN-γ/IL-4 in the acute stage , subacute stage and restoration stage were gradually increased ( P<0.05), the level of IL-4 was gradually decreased ( P<0.05), but the levels of serum IFN-γ, IL-4 and IFN-γ/IL-4 in the restoration stage and the control group had no significant differences ( P>0.05). Conclusions:Among the children with KD in acute stage, serum level of IFN-γ is decreased while serum IL-4 level is increased, and Th1/Th2 balance shifts to Th2. Along with the stabilization of disease, the levels of serum IFN-γ and IL-4 are normalized, and Th1/Th2 balance presents a recovering trend and they almost recover to normal after entering the restoration stage.

A preliminary study on the relationship between in vivo metal implants and acquired palmoplantar keratodermas / 中华皮肤科杂志

Objective:To investigate clinical characteristics of acquired palmoplantar keratodermas (PPK) and the effect of metal implants (MIs) on plasma cytokines in patients with PPK.Methods:An observational study was conducted. Patients with acquired PPK were collected from Department of Dermatology, General Hospital of Eastern Theater Command from June 2020 to December 2021, and health checkup examinees during the same period served as controls. The PPK area and severity index was evaluated according to the eczema area and severity index, metal allergens were detected by metal patch test (MPT) , and plasma levels of cytokines, including tumor necrosis factor (TNF) -α, interferon (IFN) -γ, interleukin (IL) -4, IL-13, IL-17A and IL-8, were detected by enzyme-linked immunosorbent assay. Non-normally distributed measurement data were expressed as median (quartile 1, quartile 3) , and analyzed by rank sum test; intergroup comparisons of enumeration data were performed by chi-square test.Results:A total of 81 PPK patients were collected, including 42 males and 39 females, and their average age was 48 years (range, 21 - 65 years) . In vivo MIs were found in 37 (45.7%) patients, including dental implants in 34, orthopedic implants in 2, and cardiac implants in 1. During the same period, 36 healthy subjects were included in the control group, and there was no significant difference in the age and gender composition between the two groups ( P > 0.05) . No significant difference was observed in the PPK area and severity index between the MI group (37 cases, 0.40 [0.27, 0.75] points) and non-MI group (44 cases, 0.38 [0.19, 0.70] points; Z = 1.21, P = 0.225) . Forty-six patients in the PPK patient group and 30 in the control group were subjected to MPT, and the positive rate of MPT was significantly higher in the patient group (14/46) than in the control group (2/30, χ2 = 6.17, P = 0.013) , and was significantly higher in the patients with in vivo MIs (10/25) than in the patients without (4/21, adjusted P < 0.017) . Plasma cytokines were detected in 36 cases in the patient group and 36 in the control group. The plasma levels of TNF-α and IFN-γ were significantly lower in the patient group (66.2 [58.7, 69.3] pg/ml, 645.0 [571.5, 681.1] pg/ml, respectively) than in the control group (71.5 [64.5, 73.9] pg/ml, 716.5 [620.4, 785.0] pg/ml, respectively; both P < 0.05) , but there was no significant difference in the levels of plasma IL-4, IL-13, IL-17A and IL-8 between the two groups (all P > 0.05) . Conclusions:Positive MPT reaction is more common in acquired PPK patients, especially in patients with in vivo MIs. There may be a correlation between metal and acquired PPK, and changes in plasma TNF-α and IFN-γ levels may be related to the onset of some acquired PPK.

Acupuncture combined with western medication for ocular myasthenia gravis: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the clinical efficacy between acupuncture combined with western medication and simple western medication for ocular myasthenia gravis (OMG), and to explore its possible mechanism.@*METHODS@#A total of 60 patients of ocular myasthenia gravis were randomized into an acupuncture combined with western medication group (30 cases, 1 case dropped off) and a western medication group (30 cases, 2 cases dropped off). Oral pyridostigmine bromide tablet and prednisone acetate tablet were given in the western medication group. On the basis of the treatment in the western medication group, Tongdu Tiaoqi acupuncture (acupuncture for unblocking the governor vessel and regulating qi ) was applied at Baihui (GV 20), Fengfu (GV 16), Hegu (LI 4), Zusanli (ST 36), etc. in the acupuncture combined with western medication group, once a day, 6 days a week. The treatment was given 8 weeks in both groups. Before and after treatment, the OMG clinical absolute score was observed, electrophysiological indexes of orbicularis oculi (value of mean jitter, percentage of jitter >55 μs and percentage of blocks) were measured by single-fiber electromyography (SFEMG), serum levels of acetylcholine receptor antibody (AChR-Ab), interferon-gamma (IFN-γ) and interleukin-4 (IL-4) were detected by ELISA method.@*RESULTS@#After treatment, the OMG clinical absolute scores, values of mean jitter, percentages of jitter >55 μs, percentages of blocks and serum levels of AChR-Ab, IFN-γ and IL-4 were decreased compared before treatment in both groups (P<0.05), and those in the acupuncture combined with western medication group were lower than the western medication group (P<0.05).@*CONCLUSION@#Acupuncture combined with western medication can effectively improve ptosis, palpebra superior fatigability, eye movement disorder and neuromuscular junction dysfunction in patients with ocular myasthenia gravis, the therapeutic effect is superior to simple western medication. Its mechanism may be related to down-regulating serum levels of AChR-Ab, IFN-γ and IL-4 and promoting the recovery of orbicularis oculi function.

A Systematic Review of Prevalence and Risk Factors of Latent Tuberculosis Infection Among Medical and Nursing Students

@#Introduction: Clinical training may expose medical and nursing students to workplace hazards comparable to those encountered by healthcare workers (HCWs). This study was designed to investigate the prevalence of latent tuberculosis infection (LTBI) and associated risk factors among medical and nursing students. Methods: Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) criteria, a systematic review was conducted utilising four electronic databases to appraise and synthesise the literature on LTBI which used the tuberculin skin test (TST) and the blood interferon-gamma release assay (IGRA). Results: Original articles published in the English language between 2010 and 2020 were included, yielding 14 relevant articles. The average prevalence of LTBI in high-burden countries was 38.2% for TST and 20.6% for the IGRA test. According to TST and IGRA findings, the average prevalence of LTBI in nations with an intermediate burden was 16.7% and 4.7 %, respectively. The average prevalence was 2.8% and 1.1% from the TST and IGRA tests for low-burden countries, respectively. A greater risk of LTBI was shown to be related with an increase in age among postgraduate medical school students, a history of Bacillus Calmette-Guerin (BCG) vaccination, origin from high-risk tuberculosis (TB) countries, increased hours of hospital exposure, a history of contact with TB cases, a high body mass index, older age group students, and a lower TB knowledge score. Conclusion: The available evidence from this review emphasised the importance of developing and implementing efficient and cost-effective TB infection-control programmes, particularly in high-burden countries and amongst students at risk.

Latent tuberculosis screening before kidney transplantation in the South of Brazil / Rastreamento de tuberculose latente antes do transplante renal no Sul do Brasil

Abstract Background: Tuberculosis (TB) is a prevalent infection after kidney transplantation (KT) in high-burden countries. Latent tuberculosis infection (LTBI) screening includes previous TB history, chest radiograph findings, and tuberculin test (TST) and/or interferon-gamma release assays (IGRAs) results. We aimed to compare our routine LTBI screening of KT candidates and living donors (LD) with their IGRA results, and evaluate if this would improve isoniazid (INH) treatment referral. Methods: We evaluated adult KT candidates and LD with complete routine LTBI screening and QuantiFERON-TB® Gold In-Tube (QFT) testing. Blood samples were collected from April 4th, 2014 to October 31st, 2018, with follow-up until October 31st, 2019. Results: There were 116 KT recipients, with 30% QFT-positive results. Positive QFT was associated with past TB history (p=0.007), positive TST (p<0.0001), residual radiographic lesions (p=0.003), and diabetes (p=0.035). There were 25 LD, 40% had positive QFT. Positive QFT was associated with a positive TST (p=0.002). Positive QFT results increased INH referral in 80%. Post-transplant TB incidence was 2.6% in a median follow-up of 2 (1-33) months. No variables were associated with post-transplant TB. TB patients had inferior, although non-significant, 5-year graft survival (66.7% vs. 76.5%) (p = 0.402). Conclusion: In the present study, the association of QFT to our routine LTBI screening incremented INH treatment referral, but there was still a high incidence of post-transplant TB, possibly related to other forms of infection, such as new exposure and donor transmission.

Resumo Histórico: Tuberculose (TB) é uma infecção relativamente comum pós-transplante renal (TR) em países com alta prevalência da doença. O rastreamento de infecção latente por tuberculose (ILTB) inclui histórico prévio de TB, achados de radiografia do tórax, resultados do teste tuberculínico (TT) e/ou de ensaio de liberação de interferon-gama (IGRAs). Nosso objetivo foi comparar nossa avaliação de rotina de candidatos ao TR e doadores vivos (DV) com seus resultados de IGRA, avaliando se aumentaria o encaminhamento para tratamento com isoniazida (INH). Métodos: Avaliamos candidatos adultos ao TR e DV com rastreamento para ILTB de rotina completo e coleta de testes QuantiFERON-TB® Gold In-Tube (QFT). Coletamos amostras sanguíneas de 4 de Abril, 2014 - 31 de Outubro, 2018, com acompanhamento até 31 de Outubro, 2019. Resultados: Avaliamos 116 receptores de TR, 30% sendo QFT-positivo. QFT positivo foi associado ao histórico prévio de TB (p=0,007), TT positivo (p<0,0001), lesões radiográficas residuais (p=0,003), diabetes (p=0,035). Avaliamos 25 DV, 40% apresentaram QFT positivo. QFT positivo foi associado a TT positivo (p=0,002). Resultados positivos do QFT aumentaram o encaminhamento para INH em 80%. A incidência de TB pós-transplante foi 2,6% em uma mediana de acompanhamento de 2 (1-33) meses. Nenhuma variável foi associada à TB pós-transplante. Pacientes com TB tiveram sobrevida do enxerto em 5 anos inferior, embora não-significativa (66,7% vs. 76,5%) (p = 0,402). Conclusão: Neste estudo, a associação do QFT à nossa avaliação de ILTB de rotina aumentou o encaminhamento para tratamento com INH, mas ainda houve alta incidência de TB pós-transplante, possivelmente relacionada a outras formas de infecção, como nova exposição e transmissão pelos doadores.

Role of folliculin in interferon-γ-mediated apoptosis of and chemokine secretion by melanocytes / 中华皮肤科杂志

Objective:To investigate the role of folliculin in apoptosis of and chemokine secretion by melanocytes mediated by interferon-γ (IFN-γ) .Methods:Normal primary melanocytes were isolated from circumcised foreskin tissues from a healthy male child, and primary vitiliginous melanocytes were isolated from normally pigmented suction-blistered epidermis from patients with vitiligo after suction blister epidermal grafting. Western blot analysis was performed to determine the folliculin protein expression in normal primary melanocytes, primary vitiliginous melanocytes and a human primary melanocyte line PIG1. PIG1 cells stimulated with 10 ng/ml IFN-γ for 48 hours served as induction group, and untreated PIG1 cells served as control group. Real-time quantitative RCR (qRT-PCR) was performed to determine the mRNA expression of folliculin, autophagy-related microtubule-associated protein 1 light chain 3 (LC3) -Ⅱ and Beclin genes, and Western blot analysis to determine the protein expression of folliculin, Beclin1 and LC3Ⅱ/Ⅰ, as well as phosphorylation levels of adenosine monophosphate-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) in the above cells. Furthermore, the melanocytes stimulated with 10 ng/ml IFN-γ for 48 hours were divided into several groups: negative control group infected with an empty lentiviral vector, folliculin inhibition group infected with a folliculin-inhibiting lentivirus, autophagy enhancement group infected with a folliculin-inhibiting lentivirus followed by 2-hour treatment with a mTOR inhibitor, autophagy inhibition group infected with a folliculin-inhibiting lentivirus followed by 2-hour treatment with an AMPK inhibitor. Then, flow cytometry was conducted to detect apoptosis of PIG1 cells, and enzyme-linked immunosorbent assay to measure the concentration of chemokines CXCL10 and CCL20 in the culture supernatant of PIG1 cells in the above groups. Measurement data were compared among multiple groups by using one-way analysis of variance, and multiple comparisons were carried out by using least significant difference- t test. Results:The relative protein expression level of folliculin significantly differed among the normal primary melanocytes (0.850 ± 0.120) , primary vitiliginous melanocytes (1.507 ± 0.170) and PIG1 cells (0.697 ± 0.130; F = 50.09, P < 0.001) , and was significantly higher in the primary vitiliginous melanocytes than in the normal primary melanocytes and PIG1 cells ( t = 4.06, 5.89, respectively, both P < 0.01) . Compared with the control group, the induction group showed significantly increased relative mRNA and protein expression levels of folliculin (both P < 0.01) , but significantly decreased relative mRNA and protein expression levels of LC3Ⅱ and Beclin (all P < 0.01) ; moreover, the induction group showed significantly decreased LC3Ⅱ/Ⅰ levels (0.72 ± 0.02) and AMPK phosphorylation levels (0.714 ± 0.023) in the PIG1 cells compared with the control group (1.13 ± 0.02, 1.176 ± 0.002, t = 7.34, 6.67, respectively, both P < 0.01) , but significantly increased mTOR phosphorylation levels (1.051 ± 0.023) compared with the control Group (0.451 ± 0.016, t = 3.81, P = 0.009) . There were significant differences in the PIG1 cell apoptosis rate and concentrations of CXCL10 and CCL20 among the control group, induction group and other treatment groups (all P < 0.001) ; specifically, the PIG1 cell apoptosis rate and concentrations of CXCL10 and CCL20 were significantly higher in the induction group than in the control group, lower in the folliculin inhibition group than in the negative control group, lower in the autophagy enhancement group than in the folliculin inhibition group, and higher in the autophagy inhibition group than in the folliculin inhibition group (all P < 0.05) . Conclusions:Folliculin is highly expressed in vitiliginous melanocytes. Folliculin expression and downstream signaling pathways are regulated by IFN-γ, and folliculin may participate in IFN-γ-mediated melanocyte apoptosis and chemokine secretion via regulating autophagy.

Correlation between interferon-γ rs2430561 single nucleotide polymorphisms and susceptibility to herpes zoster / 中华皮肤科杂志

Objective:To investigate the correlation of interferon (IFN) -γ rs2430561 single nucleotide polymorphisms (SNPs) and serum IFN-γ levels with susceptibility to herpes zoster.Methods:Blood samples were collected from 74 patients with herpes zoster and 40 healthy controls in General Hospital of Southern Theatre Command and the Fifth People Hospital of Hainan Province from November 2019 to November 2020. PCR and Sagner resequencing were conducted to detect the IFN-γ rs2430561 SNPs in the subjects, fluorescence-based quantitative PCR was performed to determine the copy number of varicella-herpes zoster virus (VZV) DNA in the serum of the patients with herpes zoster, and enzyme-linked immunosorbent assay was conducted to detect the serum IFN-γ level. Measurement data were compared by using t test or non-parametric test, and enumeration data by using chi-square test or Fisher′s exact test. Results:As rs2430561 genotyping showed, there were 4 patients with AA genotype, 37 with TT genotype and 33 with TA genotype in the herpes zoster group, as well as 13 subjects with TA genotype and 27 with TT genotype in the healthy control group, and the frequency of A allele of rs2430561 was significantly higher in the herpes zoster group (27.70%) than in the control group (16.25%, P=0.036) . The serum IFN-γ level ( M[ P25, P75]) was significantly lower in the herpes zoster group (33.45[0.80, 95.01]pg/ml) than in the control group (67.83[2.74, 318.35]pg/ml, U=1 822, P=0.028) . The VZV DNA copy number (expressed as a logarithm to the base of 10) per milliliter was 3.23 ± 0.71 in the serum of the patients with herpes zoster, and the serum IFN-γ level was negatively correlated with the VZV DNA copy number ( r=-0.302, P=0.009) . Conclusion:The carriage of rs2430561 A allele may affect the expression of IFN-γ, leading to higher susceptibility to herpes zoster.

Effects of intrathecal morphine and fentanyl on IFN-γ levels in hippocampus and plasma of rats with incisional pain / 中华麻醉学杂志

Objective:To evaluate the effects of intrathecal morphine and fentanyl on interferon (IFN)-γ levels in hippocampus and plasma of rats with incisional pain.Methods:Ninety-six healthy male Sprague-Dawley rats in which intrathecal catheters were successfully inserted, weighing 180-220 g, aged 6-8 weeks, were divided into 4 groups ( n=24 each) using a random number table method: normal saline group (group NS), incisional pain group (group P), morphine and fentanyl group (group MF) and morphine and fentanyl with incisional pain group (group MFP). Incisional pain model was established in group P and group MFP.At 20 min before the model was established, a 50 μl mixture of morphine 5 μg/kg and fentanyl 0.25 μg/kg was intrathecally injected in group MF and group MFP, while normal saline 50 μl was injected intrathecally in group NS and group P. At 24 h before establishment of the model (T 0) and at 1, 6, 24, 48 and 72 h after establishment of the model (T 1-5), 6 mice were randomly selected from each group for determination of the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL). The animals were sacrificed and hippocampal tissues and blood samples from the inferior vena cava were collected for determination of IFN-γ levels in hippocampal tissues and plasma (by enzyme-linked immunosorbent assay). Results:Compared with group NS, MWT was significantly decreased and TWL was shortened at T 1-5, and IFN-γ concentration in plasma was decreased at T 2, 3 and T 5 in group P, MWT was increased and TWL was prolonged at T 1-3 in group MF, MWT was decreased and TWL was shortened at T 1-3 in group MFP, and IFN-γ concentration in plasma was decreased at T 2 in MF and MFP groups ( P<0.05). Compared with group P, MWT was increased, TWL was prolonged at T 1-5, and IFN-γ concentration in plasma was increased at T 2, 3 and T 5 in MF and MFP groups ( P<0.05). Compared with group MF, MWT was decreased and TWL was shortened at T 1-4, and IFN-γ concentration in plasma was increased at T 2 and T 3 in MFP ( P<0.05). There was no significant difference in IFN-γ concentration at each time point among the 4 groups ( P>0.05). Conclusion:Intrathecal morphine and fentanyl can increase plasma IFN-γ concentration, and improve peripheral immunosuppression.

Research progress on the correlation between interleukin-18 and chronic periodontitis / 口腔疾病防治

@#Chronic periodontitis is an infectious disease caused by plaque as the initiating factor. Clinically, it manifests as irreversible loss of hard tissue, leading to the destruction of surrounding periodontal tissue, including the deep periodontal pocket, loss of attachment, and finally, tooth loss. Interleukin-18 (IL-18) can promote inflammation and regulate immune function and plays an important role in mediating the host immune response and inflammatory response. An increase in IL-18 in vivo can induce the production of interferon and inflammatory factors, such as interleukin, tumor necrosis factor and matrix metalloproteinase, thus mediating the dual reaction of immunity and inflammation. These inflammatory factors are involved in the occurrence and development of chronic periodontitis. Many clinical studies have shown that the levels of IL-18 in serum, saliva, gingival crevicular fluid and gingival tissue samples of patients with chronic periodontitis may be positively correlated with the severity of periodontitis; however, as a candidate gene, IL-18 is involved in the susceptibility polymorphism of periodontitis. Understanding how to quantify the level of IL-18 in clinical studies and apply it to diagnostic tools and new sites identified by new methods (genome-wide association studies and omics research) will also deepen our understanding of the pathogenesis of IL-18 in chronic periodontitis and provide new ideas for future precision medicine and the formulation of personalized programs. In this paper, the structure, biological function and association between IL-18 and periodontitis are reviewed.

Research progress on the relationship between IL-37 and periodontitis / 口腔疾病防治

@#Periodontitis is the inflammation of periodontal tissue caused by dental plaque, which absorbs the alveolar bone and cementum. The immune response triggered by CD4+T cells is the key factor for the aggravation of periodontitis. The activation of dendritic cells and the receptor activator of the NF-κB ligand (RANKL) pathway is an important link in the alveolar bone resorption of periodontal tissue. Pro-inflammatory factors such as interferon-γ (IFN-γ), tumor necrosis factor (TNF-α) and interleukin-1β (IL-1β) also play important roles in the development of periodontitis. Interleukin-37(IL-37), which is a newly discovered cytokine in the IL-1 family, has five shear variants from a to e, among which the clover β-structure encoded by exon 4 plays an important role in the binding of cytokines and the corresponding receptors. IL-37 has strong anti-inflammatory and inhibition of autoimmunity, can enter the nucleus with the help of caspase-1 and bind with Smad proteins to regulate the transcription of pro-inflammatory genes. Extracellular IL-37 can bind to IL-18 binding protein and inhibit the production of pro-inflammatory factors. IL-37 can inhibit the progression of periodontitis by inhibiting the RANKL signaling pathway, inhibiting the proliferation and differentiation of dendritic cells and CD4+T cells, binding to Smad proteins, and releasing pro-inflammatory factors such as IFN-γ and TNF-α. The IL-37 concentration in periodontal tissue can indicate the progression of periodontitis. Few studies have described the interaction between the anti-inflammatory factor IL-37 and periodontitis. Thus, in this paper, the structure and function of IL-37 and the related factors between IL-37 and periodontitis will be reviewed.

Obesidad e inflamación en estudiantes de una universidad pública colombiana / Obesity and inflammation in students of a Colombian public university

RESUMEN Objetivo Describir la relación entre obesidad y variables proinflamatorias en la población estudiantil de la Universidad del Quindío. Métodos Estudio descriptivo de corte transversal en el cual participaron 155 estudiantes. En primer lugar se midió la glicemia -por métodos convencionales-, la insulinemia y las citocinas -mediante la prueba ELISA-; también se calculó IMC e índice HOMA-IR. Luego, esta información se analizó en el software Statgraphics con el fin de establecer relaciones entre las variables a través de regresión múltiple. Así, se llegó a una diferencia estadísticamente significativa cuando el valor de p fue ≤0,05. Resultados La prevalencia de obesidad por IMC fue de 4,6%; la de sobrepeso, 20,5%; y la de obesidad abdominal por perímetro de cintura fue de 18,5%. El promedio de insulinemia fue de 8,5 pUI/ml±6,4 y el índice HOMA-IR, de 1,7. El 18,5% presentaron resistencia a la insulina. El interferón gamma se encontró en 4,6±12,0 pg/mL; la IL-6, en 5,1±15,5 pg/mL; y el TNF alfa, en 3,2±11,1 pg/mL. La mayor variación explicada (R2) la presentan la insulinemia, el índice cintura/cadera y el índice Homa-IR. La IL-6 fue la citocina que presentó la mayor variación explicada. Conclusiones Se evidencia que existe una relación entre citocinas inflamatorias como la IL-6 con el índice cintura/cadera, la insulinemia y el índice Homa-IR y la presión diastólica.

ABSTRACT Objective To describe the relationship between obesity and proinflammatory variables in the student population of the University of Quindío. Methods A cross-sectional descriptive study in which 155 students participated. Firstly, the glycemia was made by conventional methods, the insulinemia and the cytokines were measured by ELISA; IMC and HOMA-IR index were calculated. Then, the information was analyzed in the Statgraphics software to establish relationships between the variables were established through multiple regression and thus statistically significant difference was considered when the p value was ≤0.05. Results The prevalence of obesity by BMI was of 4.6%, of overweight 20.5% and of abdominal obesity by waist circumference was of 18.5%. The average insulinemia was 8.5 pUI/ml ±6.4 and the HOMA-IR index was 1.75. 18.5% had insulin resistance. Interferon gamma was found at 4.6±12.0 pg/mL, IL-6 at 5.1±15.5 pg/mL and TNF alpha at 3.2±11.1 pg/mL. The greatest variation explained (R2) is presented by insulinemia, the waist/hip index and the Homa-IR index. IL-6 was the cytokine that presented the greatest variation explained. Conclusions There is a relationship between inflammatory cytokines such as IL-6 with the waist/hip index, diastolic pressure, insulinemia and the Homa-IR index.

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