Th17 lymphocytes have been recently identified as a novel subset of CD4+ cells. It has been defined that IL-17,the main product of Th17, plays an important role in immunity against parasitic infection. There is a two-way influence between Th17 and cytokine network: on one hand Th17 consummates cytokine network, on the other hand many cytokines regulate Th17's activity in parasitic infection. In the anti-parasitic infection process, Th17 cells protect host or promote inflammation, even cause immune pathogenesis in different cases, which comprise host's immune state, the burden of parasitic infection, as well as the treatment.

Objective To investigate the role of serum and glucocorticoid regulated protein kinase (SGK) 1 in the inflammatory responses mediated by toll like receptors.Methods Mice were injected with lipopolysaccharide (LPS,1 mg/kg) 2 h after the pretreatment of EMD638683 (10 mg/kg) or phosphate buffered saline (PBS) as control.At the time points of 3 and 24 h,pro-inflammatory cytokines (interleukin [IL]-6,IL-12 and tumor necrosis factor [TNF]-α) in serum were measured using enzymelinked immunosorbent assay (ELISA).Livers and lung were harvested at 6 h and 24 h after the injection of LPS,embedded by optimum cutting temperature (OCT) and then stained with hematoxylin and eosin (HE).Peripheral blood mononuelear cell (PBMC) were isolated and stimulated by LPS with or without the pretreatment of EMD or LY294002.Cytokines (IL-6,IL-12 and TNF-α) were measured using ELISA.IKKα/β,IKBα and nuclear factor (NF)-κB p65 were detected by Western bolt.Data were analyzed by one way analysis of variance.Results In the model of LPS-induced endotoxin sepsis,inhibition of SGK1 induced secretion of pro-inflammatory cytokine (IL-6 [t=3.007,P＜0.05],IL-12[t=4.413,P＜0.05] and TNF-α[t=5.403,P＜0.05]),increased inflammatory cells infikration into the liver and lung within 6 h,and induced serious multiple organ damage with collapse of alveoli and fatty degeneration of liver.After 24 h,pharmacological inhibition of SGK1 with EMD638683 increased proinflammatory cytokine (IL-6 [t=18.540,P＜0.01],IL-12[t=16.520,P＜0.01] and TNF-α[t=34.880,P＜0.01]) production in human PBMC upon LPS stimulation and inhibited the phosphorylation of IKKα/ β/IKBα and nuclear factor (NF)-κB p65.Conclusions SGK1 suppresses the toll like receptor 4 mediated inflammatory responses via NF-κB.

Objective To estimate the influence of palmitic acid (PA) on the proliferation of and production of inflammatory mediators by a human keratinocyte line HaCaT.Methods Cultured HaCaT cells were treated with PA of eight concentrations (0-200 μmol/L) for 3-24 hours followed by the evaluation of cell proliferation by using the cell counting kit-8.According to the proliferation assay,four concentrations (75,100,125,150 μmol/L) of PA were selected and used to treat HaCaT cells for 24 hours,then,fluorescence-based immunohistochemical staining was performed to observe the nuclear translocation of nuclear factor (NF)-κB p65,enzyme linked immunosorbent assay (ELISA) to determine the level of interleukin (IL)-6 in the supernatant of culture medium,real-time PCR to detect the mRNA expressions of peroxisome proliferator-activated receptor oα (PPARα) and IL-6,and Western blot to quantify the protein expressions of PPARα as well as total and nuclear NF-κB p65.Those HaCaT cells receiving no treatment served as the control group.Statistical analysis was carried out by one-factor analysis of variance using the GraphPad Prism 5.0 software.Results The HaCaT cells treated with PA of 50-175 μ mol/L showed accelerated proliferation compared with the control HaCaT cells (all P ＜ 0.05).PA from 75 to 150 μmol/L enhanced the nuclear translocation of NF-κB p65,mRNA and protein expressions of PPARα,as well as the mRNA expression and supernatant level of IL-6 in a dose-dependent manner.The relative expression level of nuclear NF-κB p65 protein was 0.4536 ± 0.0173,0.5184 ± 0.0206,0.5333 ± 0.0231,0.6160 ± 0.0297,and the supernatant level of IL-6 was (31.5677 ± 0.2268),(32.3773 ± 0.4156),(32.9837 ± 0.0029) and (33.6890 ± 0.0936) ng/L,in HaCaT cells treated with PA of 75,100,125 and 150 μmol/L,respectively,compared to 0.3237 ± 0.0114 (all P ＜ 0.01) and (30.4577 ± 0.5131) ng/L (all P ＜ 0.01) in the control HaCaT cells,respectively.Conclusions PA can accelerate the proliferation of HaCaT cells,enhance NF-κB nuclear transfer,PPARα expression and IL-6 secretion in a dose-dependent manner within a certain concentration range,and may exert a promoting role in the activation and expression of some inflammatory factors.

Objective To observe the effect of sirolimus,an autophagy enhancer,on premature senescence in fibroblasts induced by repeated exposure to a subtoxic dose of ultraviolet B (UVB).Methods Skin fibroblasts from foreskin tissue of healthy adolescents were classified into six groups:control group cultured in Dulbecco's modified Eagles' medium (DMEM) containing 1％ calf serum,UVB group receiving UVB irradiation only,sirolimus group treated with sirolimus of 10 mg/L (added after daily exchange of culture medium),and three combined groups receiving UVB irradiation immediately followed by overnight treatment with sirolimus of 0.1,1.0 and 10.0 mg/L respectively.UVB irradiation was given at a dose of 10 mJ/cm2 once a day for five successive days.After five days of treatment,cell counting kit-8 (CCK-8) was used to evaluate cell viability,β-galactosidase staining to detect senescent ceils,Western blot to quantify the expressions of p53,LC3-B and beclin 1 in these fibroblasts.Autophagy level was determined by acridine orange staining followed by fluorescence microscopy and transmission electron microscopy.Data were processed by the SPSS 16.0 software,and statistical analysis was done by one-way analysis of variance,t test and least significance difference.Results Sirolimus significantly increased the proliferative activity of fibroblasts in a dose-dependent manner,with the absorbance value at 450 nm being 0.27 ± 0.02,0.36 ± 0.04 and 0.39 ± 0.04 for fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L respectively,compared to 0.26 + 0.01 for fibroblasts irradiated with UVB only (all P ＜ 0.05).Significant differences were also observed between the fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L and those irradiated with UVB only in the percentage of β-galactosidasepositive fibroblasts (92.50％ ± 0.34％,42.40％ ± 0.53％ and 6.20％ ± 0.39％ vs.95.10％ ± 0.32％,all P ＜ 0.05)and intracellular intensity of acridine orange-induced fluorescence (36.43 ± 0.24,45.25 ± 0.33 and 48.69 ± 0.37 vs.33.99 ± 0.32,all P ＜ 0.05).Moreover,the expressions of p53,LC3-B and beclin 1 in the three combined groups differed significantly from those in the UVB group (all P ＜ 0.05).Conclusion Sirolimus can inhibit UVBinduced premature senescence likely via upregulation of autophagy in fibroblasts.

Atopic dermatitis (AD) is a highly heterogeneous skin disease. The subtypes of AD classified by age, severity and inflammatory patterns have been widely accepted previously; however, the heterogeneity in skin lesions across different body sites has been rarely addressed. Most recently, it has been found that the efficacy of dupilumab varies across different body sites, suggesting different patterns of inflammation in skin lesions at different body sites. Thus, this article proposes the concept of heterogeneity across body sites in AD, summarizes cell biological features and microbiome at different skin sites under physiological conditions, analyzes clinical manifestations of, multi-omic study results from and treatment response of AD at different sites, and discusses pathogenesis of AD, providing a basis for individualized therapy of AD.

BACKGROUND: Atopic dermatitis (AD) is recognized as a common inflammatory skin disease and frequently occurred in Asian and Black individuals. OBJECTIVE: Since the limitation of dataset associated with human severe AD, this study aimed to screen potential novel biomarkers involved in mild AD. METHODS: Expression profile data (GSE75890) were obtained from the database of Gene Expression Omnibus. Using limma package, the differentially expressed genes (DEGs) between samples from AD and healthy control were selected. Furthermore, function analysis was conducted. Meanwhile, the protein-protein interaction (PPI) network and transcription factor (TF)-miRNA-target regulatory network were constructed. And quantitative real-time polymerase chain reaction (qRT-PCR) was used to validate the expressions patterns of key genes. RESULTS: In total, 285 DEGs including 214 upregulated and 71 downregulated genes were identified between samples from two groups. The upregulated DEGs were mainly involved in nine pathways, such as hematopoietic cell lineage, pertussis, p53 signaling pathway, staphylococcus aureus infection, and cell cycle, while tight junction was the only pathway enriched by the downregulated DEGs. Cyclin B (CCNB)1, CCNB2, cyclin A (CCNA)2, C-X-C motif chemokine ligand (CXCL)10, and CXCL9 were key nodes in PPI network. The TF-miRNA-target gene regulatory network focused on miRNAs such as miR-106b, miR-106a, and miR-17, TFs such as nuclear factor kappa B subunit 1, RELA proto-oncogene, Sp1 transcription factor, and genes such as matrix metallopeptidase 9, peroxisome proliferator activated receptor gamma , and serpin family E member 1. Moreover, the upregulation of these genes, including CCNB1, CCNB2, CCNA2, CXCL10, and CXCL9 were confirmed by qRT-PCR. CONCLUSION CCNB1, CCNB2, CCNA2, and CXCL9 might be novel markers of mild AD. miR-106b and miR-17 may involve in regulation of immune response in AD patients.

ObjectiveTo investigate the mechanism of action of miR-196b in regulating the growth and apoptosis of hepatoma cells by targeting nuclear apoptosis-inducing factor 1 (NAIF1). MethodsReal-time PCR was used to measure the expression of miR-196b in hepatoma HuH-7, SNU-449, HepG2, and SMCC7721 cells versus normal human HL7702 hepatocytes. The hepatoma HepG2 cells were collected and divided into Control group (blank control), Anti-NC group (transfected with inhibitor control), Anti-miR-196b group (transfected with miR-196b inhibitor), si-NC group (transfected with siRNA control), si-NAIF1 group (transfected with NAIF1 siRNA), Anti-miR-196b+si-NAIF1 group (co-transfected with miR-196b inhibitor and NAIF1 siRNA), and Anti-miR-196b+si-NC group (co-transfected with miR-196b inhibitor and siRNA control). MTT assay was used to measure the change in proliferation, plate colony formation assay was used to measure colony formation ability, flow cytometry was used to measure cell apoptosis, and Western blot was used to measure the protein expression of Bax and C-caspase-3. Target gene prediction software predicted that NAIF1 might be a target gene of miR-196b, and the luciferase reporting system was used to identify the targeting relationship. The t-test was used for comparison of continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the SNK-q test was used for further comparison between two groups. ResultsThere was a significant difference in the expression level of miR-196b between hepatoma HuH-7, SNU-449, HepG2, and SMCC7721 cells and normal human HL7702 hepatocytes (1.85±0.16/1.63±012/2.36±0.25/1.92±0.13 vs 1.00±0.09, F=29.05, P＜0.001). Compared with the Anti-NC group, the Anti-miR-196b group had significant reductions in the expression level of miR-196b (0.42±0.03 vs 1.02±0.10, P＜0.05), cell proliferation (0.20±0.02 vs 0.30±0.05, P＜0.05), and colony formation ability (64.35±6.97 vs 119.54±11.82, P＜0.05) and significant increases in apoptosis rate (22.30%±2.09% vs 4.26%±0.35%, P＜0.05) and relative protein expression of Bax (0.69±0.08 vs 0.30±0.05, P＜0.05) and C-caspase-3 (0.63±0.05 vs 0.21±0.04, P＜0.05). Compared with the si-NC group, the si-NAIF1 group had significant increases in proliferation ability (0.46±0.05 vs 0.31±0.04, P＜0.05) and colony formation ability (138.92±9.66 vs 118.47±838, P＜0.05) and significant reductions in apoptosis rate (4.12%±0.40% vs 1.23%±0.12%, P＜0.05), NAIF1 (0.10±0.01 vs 0.17±0.02, P＜0.05), and protein expression of Bax (0.18±0.02 vs 0.29±0.03, P＜0.05) and C-caspase-3 (0.12±0.01 vs 020±0.03, P＜0.05). Compared with the Anti-miR-196b+si-NC group, the Anti-miR-196b+si-NAIF1 group had significant increases in proliferation ability (0.28±0.02 vs 0.21±0.03, P＜0.05) and colony formation ability (97.12±8.23 vs 66.35±5.20, P＜0.05) and significant reductions in apoptosis rate (9.60%±1.11% vs 21.14%±1.32%, P＜0.05), NAIF1 (0.30±0.04 vs 0.52±0.06, P＜0.05), and protein expression of Bax (0.28±0.03 vs 0.67±0.06, P＜0.05) and C-caspase-3 (0.22±0.05 vs 0.60±004, P＜0.05). ConclusionDownregulation of miR-196b can inhibit the growth and induce the apoptosis of hepatoma cells via negative regulation of NAIF1.

Objective:To analyze blood transfusion and prognostic factors of extracorporeal membrane pulmonary oxygenation (ECMO) for the treatment of respiratory and circulatory failure.Methods:The clinical data of 80 patients with respiratory and circulatory failure who received treatment in Huangshi Central Hospital from March 2016 to July 2021 were retrospectively analyzed. According to 28-day prognosis, these patients were divided into death group ( n = 44) and survival group ( n = 36). The general data, blood transfusion during the process of ECMO, vital signs, laboratory indicators, ventilation time, and length of hospital stay were compared between the two groups. The factors affecting death during the process of ECMO were analyzed. Results:There were no significant differences in sex, age, body mass index, complications, the cause of respiratory and circulatory failure, and the mode of ECMO between the two groups (all P > 0.05). Preoperative Acute Physiology and Chronic Health Evaluation II score, creatinine, procalcitonin and lactic acid levels in the survival group were (22.36 ± 3.71) points, (79.17 ± 9.29) μmol/L, (2.77 ± 0.79) ng/L, (2.74 ± 0.36) mmol/L, respectively, which were significantly lower than (34.27 ± 4.98) points, (94.16 ± 10.23) μmol/L, (3.69 ± 1.10) ng/L, (5.18 ± 0.42) mmol/L, respectively in the death group ( t = -11.89, -6.79, -5.62, -27.53, all P < 0.001). There were no significant differences in preoperative respiratory frequency, diastolic pressure, systolic pressure, heart rate, oxygenation index (PaO 2/FiO 2) and C-reactive protein between the two groups (all P > 0.05). The volume of blood transfused on the day of undergoing ECMO, the volume of blood transfused on the day of withdrawing ECMO, the volume of blood transfused during the whole process of ECMO, duration of ventilation, and the incidence of complications related to ECMO were(98.74 ± 16.28) mL, (37.23 ± 10.36) mL, (398.79 ± 67.81) mL, (210.39 ± 20.21) hours, 38.89% (14/36), respectively, which were significantly lower than (160.17 ± 23.14) mL, (48.26 ± 12.25) mL, (600.23 ± 70.12) mL, (320.14 ± 18.21) hours, 79.55% (35/44), respectively in the death group ( t = -13.43, -4.29, 4.94, 25.25, χ2 = 13.79, all P < 0.001). The length of hospital stay in the survival group was longer than that in the death group [(20.14 ± 5.36) days vs. (14.17 ± 4.23) days, t = 5.56, P < 0.001). Acute Physiology and Chronic Health Evaluation II score, procalcition level, the volume of blood transfused on the day of ECMO, duration of ventilation, and the volume of blood transfused during the whole process of ECMO are risk factors for death after ECMO, while length of hospital stay is a protective factor for ECMO. Conclusion:Preoperative evaluation of Acute Physiology and Chronic Health Evaluation II score, continuous blood transfusion during the whole process of ECMO, grasping the opportunity of ventilation and preventing against complications of ECMO are the keys to increasing the survival rate of patients with respiratory and circulatory failure.

Objective:To investigate the efficacy of phenolamine in the treatment of sepsis-induced myocardial dysfunction and its effect on cardiac function, myocardial injury index, and hemodynamics in patients.Methods:The clinical data of 79 patients with sepsis-induced myocardial dysfunction who received treatment in Huangshi Central Hospital, Edong Healthcare Group from February 2017 to February 2020 were retrospectively analyzed. These patients were divided into a control group (without phenolamine treatment, n = 41) and an observation group (with phenolamine treatment, n = 38) according to whether they received phenolamine treatment or not. Clinical efficacy, cardiac function, myocardial injury index, and hemodynamic index pre- and post-treatment were compared between the two groups. Results:There was no significant difference in 28-day mortality rate between the two groups ( P > 0.05). Intensive care unit length of stay and mechanical ventilation duration in the observation group were (9.33 ± 3.52) days and 83.00 (28.50, 138.00) hours, which were significantly shorter than (12.17 ± 4.15) days and 111.00 (47.50, 169.00) hours in the control group ( t = 3.26, Z = -2.27, both P < 0.05). The response rate in the observation group was significantly higher than that in the control group [81.58% (31/38) vs. 60.98% (25/41), χ2 = 4.05, P < 0.05]. After 7 days of treatment, the left ventricular ejection fraction in each group was significantly increased, and the left ventricular end-diastolic diameter and left ventricular end-systolic diameter in each group were significantly decreased compared with before treatment (all P < 0.05). After 7 days of treatment, the left ventricular ejection fraction in the observation group was significantly higher than that in the control group ( t = 3.29, P < 0.05), and left ventricular end-diastolic diameter and left ventricular end-systolic diameter were significantly lower than those in the control group ( t = 5.94, 11.21, both P < 0.05). N-terminal pro-brain natriuretic peptide and cardiac troponin I levels in each group were significantly decreased with time (both P < 0.05). At 24 and 72 hours and 7 days after treatment, N-terminal pro-brain natriuretic peptide and cardiac troponin I levels in the observation group were significantly lower than those in the control group (both P < 0.05). After 7 days of treatment, heart rate in each group decreased significantly compared with that before treatment (both P < 0.05), mean arterial pressure, cardiac index, and stroke output index in each group increased significantly compared with those before treatment (all P < 0.05). After 7 days of treatment, heart rate in the observation group was significantly lower than that in the control group ( t = 4.90, P < 0.05), and mean arterial pressure, cardiac index, and stroke output index in the observation group were significantly higher than those in the control group ( t = 4.37, 3.23, 6.01, all P < 0.05). Conclusion:Phentolamine can improve hemodynamics, reduce myocardial injury and improve cardiac function in patients with sepsis-induced myocardial dysfunction.