Minocycline induces tolerance to dendritic cell production probably by targeting the SOCS1/ TLR4/NF-κB signaling pathway.

OBJECTIVE: Dendritic cells (DCs) are professional antigen-presenting cells that play a key role in maintaining peripheral immune tolerance. The use of tolerogenic DCs (tolDCs), i.e., semi-mature DCs that express co-stimulatory molecules but not pro-inflammatory cytokines, has been proposed. However, the mechanism of tolDCs induced by minocycline is still unclear. Our previous bioinformatics analyses based on multiple databases suggested that the suppressor of cytokine signaling 1/Toll-like receptor 4/NF-κB (SOCS1/TLR4/NF-κB) signal pathway was associated with DCs maturation. Thus, we studied whether minocycline could induce DC tolerance through this pathway. METHODS: A search for potential targets was carried out through public databases, and pathway analysis was performed on these potential targets to obtain pathways relevant to the experiment. Flow cytometry was used to detect the expression of DC surface markers CD11c, CD86, and CD80, and major histocompatibility complex II. The secretion of interleukin (IL)-12p70, tumor necrosis factor alpha (TNF- α), and IL-10 in the DC supernatant was detected by enzyme-linked immunoassay. The ability of three groups (Ctrl-DCs, Mino-DCs, and LPS-DCs) of DCs to stimulate allogeneic CD4+ T cells was analyzed using a mixed lymphocyte reaction assay. Western blotting was used to detect the expression of TLR4, NF-κB-p65, NF-κB-p-p65, IκB-α, and SOCS1 proteins. RESULTS: The hub gene plays a vital role in biological processes; in related pathways, the regulation of other genes is often affected by it. The SOCS1/TLR4/NF-κB signaling pathway was further validated by searching for potential targets through public databases to obtain relevant pathways. The minocycline-induced tolDCs showed characteristics of semi-mature DCs. Moreover, the IL-12p70 and TNF-α levels in the minocycline-stimulated DC group (Mino-DC group) were lower than those in the lipopolysaccharide (LPS)-DC group, and the IL-10 levels were higher in the Mino-DC group than in the LPS-DC and control DC groups. In addition, the Mino-DC group had decreased protein expression levels of TLR4 and NF-κB-p65 and upregulated protein levels of NF-κB-p-p65, IκB-α, and SOCS1 compared with the other groups. CONCLUSION: The results of this study indicate that minocycline could improve the tolerance of DCs probably by blocking the SOCS1/TLR4/NF-κB signaling pathway.

Microfluidic-based exosome isolation and highly sensitive aptamer exosome membrane protein detection for lung cancer diagnosis.

Non-invasive methods of detecting cancer by circulating exosomes are challenged by inefficient purification and identification. This study hereby proposed an automated centrifugal microfluidic disc system combined with functionalized membranes (Exo-CMDS) to isolate and enrich exosomes, which will then be processed by a novel aptamer fluorescence system (Exo-AFS) in order to detect the exosome surface proteins in an effective manner. Exo-CMDS features in highly qualified yields with optimal exosomal concentration of 5.1 × 109 particles/mL from trace amount of blood samples (<300 µL) in only 8 min, which truly accomplishes the exosome isolation and purification in one-step methods. Meanwhile, the limit of detection (LOD) of PD-L1 in Exo-AFS reaches as low as 1.58 × 105 particles/mL. In the trial of clinical samples, the diagnostic accuracy of lung cancer achieves 91% (95% CI: 79%-96%) in contrast to the exosome ELISA (area under the curve: 0.9378 versus 0.8733; 30 patients). Exo-CMDS and Exo-AFS display the precedence in the aspects of inexpensiveness, celerity, purity, sensitivity and specificity when compared with the traditional techniques. Such assays potentially grant a practicable way of detecting inchoate cancers and guiding immunotherapy in clinic.

Reference dose prediction by using CDK molecular descriptors: A non-experimental method.

Reference dose (RfD) is an estimate of a daily dose that individual can be exposed chronically without obvious deleterious effects during a lifetime. In the area of toxicology, researchers always use the traditional approach by employing NOAEL/LOAEL or the benchmark dose (BMD) and other dose-response approaches to estimate RfD. These methods have, despite their typicalness, certain limitations. In this study, we present a novel method of the estimation of reference dose without experiments. The information of the organic chemicals is available from the Integrated Risk Information System (IRIS) of USEPA. Molecular descriptors for each molecular structure were calculated by an integrated platform, and the chemicals were classified into four categories based on molecular similarity: 128 contained benzene rings, 47 were heteroaromatics, 104 contained halogen substituents and 44 were halogenated aliphatic hydrocarbons. The predictive model of RfD was constructed by the multiple linear stepwise regression (MLR) method. Approximately 95% and 82% of the data points differ by less than 10-fold and 5-fold between the predicted values and the true values respectively. The non-experimental method improves the estimation efficiency and has a certain reference value to predict.

Perivascular space is associated with brain atrophy in patients with multiple sclerosis.

BACKGROUND: Perivascular space (PVS) is associated with neurodegenerative and neuroimmune diseases. Multiple sclerosis (MS) is traditionally a neuroimmune disease. However, studies show neurodegeneration also plays a vital role in MS. At present, most studies conclude severer PVS in MS is an imaging marker of neuroinflammation, while a 7T MRI study suggests that PVS in MS is associated with neurodegeneration. METHODS: In this study, 82 MS patients (n=82) and 32 healthy controls (n=32) were enrolled. The following indexes were measured: the number, size and distribution of PVS, the PVS score, corpus callosum index (CCI), corpus callosum area (CCA), the ratio of the corpus callosum to the cranium (CCR), aligned third ventricle width (a3VW), and unaligned third ventricle width (u3VW). RESULTS: The PVS score (4 vs. 3, P=0.041), PVSs number (103.280±45.107 vs. 87.625±30.139, P=0.035), and enlarged perivascular spaces (EPVSs) number (9 vs. 1, P<0.001) of MS patients were significantly higher than in the healthy controls. PVSs number (23.5 vs. 13) and EPVSs number (1 vs. 0) in the basal ganglia (BG), and EPVSs number (3 vs. 0) in centrum semiovale (CSO) of MS patients were significantly higher than in the healthy controls, P<0.001. In MS patients, PVS was correlated with age and hypertension but not to the extended disability status scale (EDSS) score and other clinical data. In MS patients, PVS score was correlated with CCA (rs=0.272; P=0.013) and the CCR (rs=0.219; P=0.048), and PVSs number was correlated with CCA (rs=0.255; P=0.021), the correlation disappeared after adjusting hypertension and age. In MS patients in remission, PVSs number was correlated with CCA (rs=0.487; P=0.019), CCR (rs=0.479; P=0.021), and PVS score was correlated with CCA (rs=0.453; P=0.03). After adjustment of hypertension and age, the total number of PVSs was correlated with CCA (rs=0.419; P=0.049). CONCLUSIONS: The PVS load in MS patients was heavier than healthy people, especially in BG and CSO. PVS was not correlated with EDSS in MS patients. The PVS of MS patients was associated with CCA and CCR, and PVSs number was independently related with CCA in MS patients in remission.

Low-dose IL-2 therapy limits the reduction in absolute numbers of circulating regulatory T cells in rheumatoid arthritis.

BACKGROUND: Circulating regulatory T cells (Tregs) are responsible for mediating immune tolerance and maintaining immunological homeostasis. Decreases in Tregs may be involved in the onset of rheumatoid arthritis (RA). Low-dose interleukin-2 (IL-2) has been considered for the treatment of inflammatory diseases mediated by T cells. This study focused on the status of circulating CD4+T subsets and the clinical feasibility of IL-2 therapies in patients with RA. METHODS: The subjects included 888 patients with RA and 100 healthy controls (HCs); 233 RA patients received IL-2 treatment with 0.5 million international units (MIU)/day from days 1 through 5. The demographic features, disease activity, and levels of CD4+T cells measured by modified flow cytometry were collected in all RA patients before and after treatment. RESULTS: RA patients had lower absolute Treg counts (but not Th17) compared with HCs, which was associated with disease activity; previously treated RA patients had the fewest circulating Tregs (p < 0.05). Patients treated with low-dose IL-2 had a three-fold increase in absolute anti-inflammatory Treg counts, as well as a two-fold increase in the other CD4+T subsets. Moreover, post-treatment levels of markers of disease activity in RA patients treated with IL-2 were significantly lower than the baseline values (p < 0.001), with no apparent side effects. CONCLUSION: Decreased absolute counts of circulating CD4+T lymphocyte subsets were observed in patients with RA. Circulating Tregs, which mediate immune tolerance, may be involved in the pathogenesis and progression of RA; however, this was ameliorated by low-dose IL-2, without obvious side effects. PLAIN LANGUAGE SUMMARY: Low-dose IL-2 treatment for rheumatoid arthritis • Circulating Tregs may be involved in the pathogenesis and progression of RA.• The absolute count of Tregs was significantly correlated with disease activity measures.• Low-dose IL-2 was able to effectively expade Tregs and help for RA patients' symptoms remission without evaluated side effects.

[Cytotoxicity of carboxymethyl chitosan zinc and peptide to human periodontal ligament cells].

PURPOSE: The purpose of this investigation was to study the cytotoxicity of carboxymethyl chitosan zinc and peptide to human periodontal ligament cells in vitro, in order to provide a reference for understanding materials' safety. METHODS: The primary cells were obtained from human periodontal tissues and cultured. The cultured cells were identified by observing the shape under microscope and by immunocytochemical method. HPDLCs were cultured in different concentrations of the tested materials extract and the activity of cells was evaluated using CCK-8 assay. The cytotoxicity grade was determined in term of the cell relative growth rates. The concentration of the tested materials extract was 100%, 75%, 50% and 25% in group 1-4, respectively. The fifth group had no materials. All statistical analysis was performed using SPSS 17.0 software package. RESULTS: The primary cells owned fibroblasts' shape. Immunocytochemical analysis showed the cells were stained positively to antibodies against vimentin, and negatively to antibodies against cytokeratin, which indicated that they were external embryo mesenchymal cell. The cell relative growth rates were more than 100%, no matter the concentrations of materials extract were. The cytotoxicity grade of CMC-Zn+-P was 0. CONCLUSIONS: CMC-Zn+-P exhibits no cytotoxicity to HPDLCs in vitro, which meets the requirements of the national standard.

[The effect of carboxymethyl chitosan zinc and peptide on IL-1,TNF-α and PGE-2 in gingival crevicular fluid of miniature pigs].

PURPOSE: This experiment was aimed at exploring whether carboxymethyl chitosan zinc and peptide （CMC-Zn(+)-P） can reduce the occurrence and development of periodontal tissue inflammation effectively by observing the change of IL-1,TNF-α and PGE-2 level in gingival crevicular fluid （GCF） before and after brushing, so as to find a new effective material in preventing and treating periodontal diseases. METHODS: Miniature pigs were selected as experimental subjects and divided into 4 groups randomly: the control group; CMC-Zn(+)-P group (material group);brushing group; brushing + CMC-Zn(+)-P group (composite group). Gingival crevicular fluid before and one month after the experiment was collected. The levels of IL-1, TNF-α and PGE-2 were examined by enzyme-linked immune-sorbent assay, while the clinical periodontal index was recorded. SPSS 18.0 software package was used for statistical analysis. RESULTS: There was no significant difference in levels of IL-1, TNF-α and PGE-2 and clinical periodontal index between the 4 groups before experiment. After one month, the levels of IL-1, TNF-α, PGE-2 in GCF had significant difference between 4 groups. The levels of IL-1, TNF-α, PGE-2 in composite group were significant lower than that of the other three groups (P<0.008).The levels of IL-1, TNF-α and PGE-2 in the material group and brushing group were significantly lower than that of the control group (P<0.008). Compared with materials group, the brushing group had significantly lower level of IL-1,significantly higher level of PGE-2 ,but no difference in the level of TNF-α.In addition, the teeth calculus index of composite group was significantly lower than that of other groups (P<0.05). CONCLUSIONS: CMC-Zn(+)-P can effectively reduce periodontal tissue inflammation and cut down the speed of deposition of dental calculus. If used cooperatively with brushing, the effect will be better.

[Study of antimicrobial activity of carboxymethyl chitosan zinc complex material against cariogenic bacteria].

PUPPOSE: To evaluate the antimicrobial activity of carboxymethyl chitosan zinc (CMCSZ) and carboxymethyl chitosan zinc-active peptide complex material (CMCSZP) on oral cariogenic bacteria. METHODS: Agar dilution method and K-B disk diffusion susceptibility agents were used to measure the antimicrobial activity of two agents against S.mutans, Lactobacillus, S.sanguinis and Actinomyces viscosus. The former method was used to measure the minimal inhibitory concentration (MIC) and latter was used to measure the inhibitory zone. The effects of pH value, temperature, light, ultraviolet and storage temperature on the active substances were investigated to determine the stability of CMCSZ and CMCSZP. SPSS17.0 software package was used for statistical analysis. RESULTS: All the bacteria were susceptible to active peptide, CMCSZ and CMCSZP with the MIC of CMCSZ/CMCSZP being 625, 1250, 1250 and 2500 mg/L, respectively. At the same concentration, the inhibitory zone of CMCSZP was significantly bigger than that of CMCSZ. Acidic conditions were conducive to increase the antimicrobial activity of CMCSZ, while the effect on CMCSZP was not significant. CMCSZ and CMCSZP exhibited good stability against light, but their antimicrobial activity gradually weakened as the bath temperature rising. In the temperature of 85 degrees centigrade, their antibacterial activity disappeared. CONCLUSIONS: CMCSZ have CMCSZP had strong antimicrobial activity against 4 kinds of cariogenic bacteria. They have good stability against light, but poor thermal stability. This study provides theoretical foundation for the application of CMCSZ/CMCSZP in prevention of cariogenic diseases.

[Comparative research on evaluation methods of quality of injury surveillance-based emergency room].

OBJECTIVE: To evaluate the quality of injury surveillance based-emergency room and find a simple, convenient and practical comprehensive evaluation method. METHODS: Seventeen indexes were extracted from influencing factors questionnaires of injury surveillance which was collected in the emergency room of a general hospital from July 2006 to March 2007, evaluated by TOPSIS (Technique for order preference by similarity to ideal solution) and GRA (grey relational analysis), given the order and compared with the practical results from injury surveillance reporting cards and emergency department logs. RESULTS: The correlation coefficient between GRA and practical results was 0.750 (P = 0.020). The correlation coefficient between TOPSIS and practical results was 0.167 (P = 0.668). The correlation coefficient between GRA and TOPSIS was 0.083 (P = 0.831). CONCLUSION: GRA is more suitable than TOPSIS for evaluation of the quality of injury surveillance based-emergency room, and it can be used to evaluate the quality of injury surveillance, and provide timely and effective basis for injury prevention strategies.

[Clinical observation on acupuncture plus cake-separated moxibustion for treatment of chronic laryngitis].

OBJECTIVE: To search for an effective therapy for chronic laryngitis. Methods Ninety cases of chronic laryngitis were randomly divided into a treatment group of 60 cases and a control group of 30 cases. The treatment group were treated by acupuncture at Futu (LI 18), Renying (ST 9), Shuitu (ST 10), etc. , and cake-separated moxibustion at Yongquan (KI 1); the control group with oral administration of Huang's Xiang Sheng Pills. Their therapeutic effects were observed after treatment of 4 weeks. The relationship of syndrome types, pathological types and therapeutic effects with the therapeutic courses in the two groups were compar treatment of chronic . RESULTS: The total effective rate was 96.7 in the treatment group and 76.6% in the control group with a very significant difference between the two groups (P < 0.01); the therapeutic effects for the yin-deficiency and lung-dryness type and the phlegm-heat accumulation type were superior to that of the control group (P < 0.05), and the therapeutic effect for the lung and spleen qi-deficiency type was similar to the control group. The therapeutic effect for chronic simple laryngitis was significantly better than that of the control group (P < 0.05); the therapeutic effect of hypertrophic laryngitis and atrophic laryngitis was similar to that of the control group. CONCLUSION: Acupuncture combined with cake-separated moxibustion has an obvious therapeutic effect on chronic laryngitis.

[Clinical observation on acupuncture for treatment of Tourette's syndrome].

OBJECTIVE: To explore an ideal program for acupuncture treatment of Tourette's syndrome (TS). METHODS: One hundred and two cases of TS were randomly divided into a treatment group of 68 cases and a control group of 34 cases. The treatment group were treated with acupuncture at Taichong (LR 3) and Hegu (LI 4) as main, and the control group with oral administration of heloperidol. Their therapeutic effects were compared, and changes of somatosensory evoked potentials (SEP) before and after treatment were investigated in the treatment group. RESULTS: After treatment for 3 courses, 56 cases were cured, 10 improved and 2 ineffective with an effective rate of 97.1% in the treatment group; and 15 cases were cured, 11 improved and 8 ineffective with an effective rate of 76.5% in the control group, with a very significant difference in the effective rate between the two groups (P < 0.001); after treatment, the abnormal rate of SEP decreased by 41.1% in the treatment group (P < 0.001). CONCLUSION: Acupuncture is a very effective therapy for TS and has a certain restoring action on mild abnormal change of SEP.