Increased CCL24 and CXCL7 levels in the cerebrospinal fluid of patients with neurosyphilis.

BACKGROUND: Monocytes are recruited into the cerebrospinal fluid (CSF) of patients with neurosyphilis, suggesting abnormal chemokine expression. We aimed to investigate the aberrant expression of chemokines in the CSF of these patients. METHODS: CSF and serum samples were collected from patients with neurosyphilis between July 2017 and June 2019 in the Dermatology Department, Second Affiliated Hospital of Zhejiang University. Differences in the expression of 38 chemokines between patients with and without neurosyphilis were detected using RayBio® Human Chemokine Antibody Array C1. CCL24 and CXCL7 levels in the patients' CSF and serum were further measured using RayBio® CCL24 and CXCL7 ELISA kits. RESULTS: Ninety-three CSF and serum samples of patients with syphilis were collected. Antibody array analysis showed that the CSF levels of CCL24 (P = .0185), CXCL7 (P < .0001), CXCL13 (P < .0001), CXCL10 (P < .0001), and CXCL8 (P < .0001) were significantly higher in patients with than without neurosyphilis. ELISA confirmed significantly higher CCL24 and CXCL7 levels in the CSF of patients with than without neurosyphilis (CCL24: 6.082 ± 1.137 pg/mL vs 1.773 ± 0.4565 pg/mL, P = .0037; CXCL7: 664.3 ± 73.19 pg/mL vs 431.1 ± 90.54 pg/mL, P = .0118). Increased CCL24 and CXCL7 expression was seen throughout all neurosyphilis stages, had moderate diagnostic efficiency for neurosyphilis, and correlated poorly with CSF cell count and Venereal Disease Research Laboratory titer. CSF CCL24 levels also correlated poorly with CSF protein concentration. CONCLUSION: Abnormally high CSF chemokines levels may play a role in the pathogenesis of neurosyphilis.

Exosomal microRNA profiles from serum and cerebrospinal fluid in neurosyphilis.

OBJECTIVE: Changes in microRNAs (miRNAs) in the cerebrospinal fluid (CSF) are associated with different neurological diseases. Since alternations of miRNAs in neurosyphilis are insufficiently investigated, we analysed miRNAs in the CSF of patients suffering from neurosyphilis. METHODS: Exosomes were isolated from serum and CSF. Levels of 44 miRNAs were determined using quantitative real-time PCR-based miRNA array. RESULTS: In patients with neurosyphilis (NSP), miR-590-5p, miR-570-3p and miR-570-5p were upregulated in the CSF and serum, when compared with patients with syphilis without neurosyphilis (SP). miR-590-5p and miR-570-3p were significantly upregulated (p<0.001). The expression of miR-21-5p was upregulated only in the CSF of NSP. Significant downregulation was observed for miR-93-3p in the CSF and serum of NSP. No statistical difference was found in the expression of miR-7-5p, miR-1307-5p, miR-203a-3p, miR-16, miR-23b-3p and miR-27b-5p in the CSF and serum of NSP and SP. CONCLUSION: For the first time, regulation profiles in miRNA in the CSF and serum were analysed in NSP. We found significant differences in upregulation and downregulation. Therefore, miRNAs may be potential biomarkers for the presence of neurosyphilis.

[Bacterial pathogenic characteristics of respiratory tract infection in children in Suzhou, China: an analysis of 14,994 cases].

OBJECTIVE: To investigate the bacterial pathogenic characteristics of respiratory tract infection in children. METHODS: The medical data from 14,994 children with respiratory tract infection who were hospitalized in Children's Hospital Affiliated to Soochow University between November 2005 and October 2014 were retrospectively reviewed. RESULTS: Among the 14,994 sputum samples from the children with respiratory tract infection, 3,947 (26.32%) had a positive bacterial culture. The most common bacterial pathogen was Streptococcus pneumonia (12.79%), followed by Haemophilus influenzae (5.02%) and Moraxella catarrhalis (2.91%). The bacterial detection rates differed significantly in different years and seasons and children of different ages (P<0.01). The children who had not taken antibacterial agents before admission had a significantly higher positive bacterial culture rate than those who had taken antibacterial agents (P<0.01). There were significant differences in the bacterial detection rate among the children with different course of disease (<1 month, 1-3 months and >3 months) (P<0.05). The detection rates of Streptococcus pneumonia, Moraxella catarrhalis and Acinetobacter baumannii showed an increased trend with a prolonged disease course (P<0.05). CONCLUSIONS: Streptococcus pneumonia is the most common bacterial pathogen causing respiratory tract infection in children, followed by Haemophilus influenzae and Moraxella catarrhalis. The detection rate of bacterial pathogens varies in different years and seasons and children of different ages. The course of the disease and application of antibacterial agents outside hospital can affect the detection rate of bacterial pathogens in children with respiratory tract infection.

Therapeutic effects of anti-B7-H3 antibody in an ovalbumin-induced mouse asthma model.

BACKGROUND: B7 molecules play a key role in regulating allergen-induced T cell activation in asthma, which may occur through T cell recruitment and T helper cell differentiation on allergen provocation. Initial studies have shown that B7-H3 (CD276), a recently identified B7 family member, plays a critical role in the development of Th2 cells. OBJECTIVE: To investigate the effects of anti-B7-H3 monoclonal antibody (mAb) in a mouse model of allergic asthma. METHODS: The asthma model was established by ovalbumin (OVA) sensitization and challenging in female BALB/c mice. Total cell numbers in bronchoalveolar lavage fluid (BALF) were determined, and the expression levels of interferon gamma (IFN-γ), interleukin (IL)-4, and IL-17 in BALF were measured by enzyme-linked immunosorbent assay. Pulmonary eosinophil infiltration and mucus production were detected by hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS), respectively. B7-H3 expression was detected by immunohistochemistry in frozen tissue sections. RESULTS: Anti-B7-H3 mAb treatment alleviated the asthmatic syndrome, decreased the levels of B7-H3-positive cells in the lung tissues, abrogated hypercellularity, eosinophil infiltration, and mucus production, and inhibited IL-4 and IL-17 production in BALF at the induction phase as compared with the immunoglobulin G (IgG) control group (P < .01). In addition, the treatment of anti-B7-H3 mAb at the induction phase could increase the expression levels of IFN-γ as compared with the IgG control group (P < .01). Anti-B7-H3 mAb treatment at the effector phase did not inhibit the asthma response. CONCLUSION: Blockade of B7-H3 signals may provide a novel therapeutic approach to the treatment of allergic asthma.

[Etiology of acute respiratory tract infection in hospitalized children in Suzhou from 2005 to 2011].

OBJECTIVE: To elucidate the etiology of acute respiratory tract infection (ARI) in hospitalized children in Suzhou from 2005 to 2011. METHODS: A total of 10 243 hospitalized children with ARI in Children's Hospital Affiliated to Soochow University from September 2005 to October 2011 were enrolled in the study. The clinical information was collected; and the nasopharyngeal aspiration fluid and serum samples were sent for multi-pathogen detection. Respiratory syncytial virus (RSV), influenza virus type A and B (IV-A, IV-B), parainfluenza virus type 1-3 (PIV-1-PIV-3) and adenovirus (ADV) were detected by direct immunofluorescence assay. Human bocavirus (HBoV), mycoplasma pneumoniae (MP) and chlamydia pneumoniae (CP) were detected by fluorescent quantitative PCR while human metapneumovirus (hMPV) was detected by reverse transcription PCR (RT-PCR). Sputum culture was applied to detect bacterial infection and quantitative ELISA was adopted to detect the specific antibodies of MP and CP. The results of the above detections were analyzed, and thereby to explore the prevalent pathogens among different aging children and the seasonal distribution and characteristics of the disease. RESULTS: At least one type of pathogen was detected in 5871 out of 10 243 hospitalized children and the overall positive rate was 57.32%; including 3326 virus samples with positive rate at 32.47% (3326/10 243), 2870 bacteria samples with positive rate at 28.02% (2870/10 243) and 2759 atypical pathogen samples,with positive rate at 26.94% (2759/10 243). MP was the most common pathogen,whose detected rate was 25.74% (2637/10 243). The median age of children with RSV (6 months) or PIV-3(8 months) infection was younger than the median age of all hospitalized children (12 months) (χ(2) = 380.992, 34.826, P < 0.05). While the median age of children with ADV (42 months), HBoV (14 months) or IV-A (24 months) infection was older than it of all hospitalized children (χ(2) = 83.583, 13.169, 18.012, P < 0.05). The median age of children with MP (30 months),streptococcus pneumoniae (17 months) or haemophilus parainfluenzae (21 months) infection was older than it of all hospitalized children (χ(2) = 728.299, 60.463, 8.803, P < 0.05). The detected rate of RSV in the groups of children aging less than 6 months, 7-12 months, 2-3 years, 4-5 years and over 6 years was separately 25.59% (840/3283), 17.05% (333/1953), 11.85% (310/2615), 6.68% (90/1347), and 2.87% (30/1045); which decreased while the age grew (χ(2) = 178.46, P < 0.01). Conversely, the positive rate of MP increased with the age growing (χ(2) = 379.21, P < 0.01). The rate in the above groups was 8.25% (271/3283), 19.46% (380/1953), 33.00% (863/2615), 41.43% (558/1347), 54.07% (565/1045), respectively. RSV and IV-A were prevalent in winter, whose detected rates were 35.73% (941/2634) and 4.44% (117/2634) respectively.hMPV infection was common in spring, with the detected rate at 10.55% (278/2634); while HBoV infection was common in summer and autumn, with the positive rate at 9.99% (149/1491) and 9.71% (98/1009). MP and CP were frequently detected in summer, up to 31.27% (819/2619) and 10.07% (43/427) respectively. RSV was the most common pathogen in bronchiolitis (33.27% (866/2603)) and MP was the most common pathogen in bronchopneumonia (26.05% (1152/4422)) and lober pneumonia (52.25% (267/511)). CONCLUSION: MP and RSV were the most common pathogens in respiratory tract infection in hospitalized children. The novel virus included hMPV and HBoV, which also played an important role in ARI. Different pathogens were prevalent in different ages; with respective seasonal distribution and characteristics.

Investigation of the relationship between community-acquired respiratory distress syndrome toxin and the high-mobility group box protein 1-toll-like receptors-myeloid differentiation factor 88 signaling pathway in Mycoplasma pneumoniae pneumonia.

BACKGROUND: In recent years, reports of refractory Mycoplasma pneumoniae pneumonia (RMPP) have gradually increased, including reports on how these conditions threaten the lives of children. However, the specific mechanism of Mycoplasma pneumoniae pneumonia (MPP) remains unclear. This study aimed to investigate the relationship between community-acquired respiratory distress syndrome toxin (CARDS TX) and High-mobility group box protein 1-Toll-like receptors-Myeloid differentiation factor 88 (HMGB1-TLRs-MyD88) in MPP and to examine the immune pathogenesis of Mycoplasma pneumoniae infection. METHODS: Children who were diagnosed with MPP and examined by bronchoscopy were included in the MPP group. Additionally, children who underwent bronchoscopy because of bronchial foreign bodies in the same period were included in the control group. Gene expression of CARDS TX, HMGB1, Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), MyD88, and cluster of differentiation 14 (CD14) in bronchoalveolar lavage fluid (BALF) were detected using real-time reverse transcription-polymerase chain reaction. Correlations between CARDS TX and HMGB1-TLRs-MyD88 were analyzed. RESULTS: CARDS TX, HMGB1, TLR2, MyD88, and CD14 mRNA expression in BALF in the MPP group was significantly higher than that in the control group (all P < 0.05). CARDS TX mRNA expression was positively correlated with HMGB1, TLR2, MyD88, and CD14 mRNA expression (all P < 0.05). Furthermore, HMGB1 mRNA expression was positively correlated with TLR2, MyD88, and CD14 mRNA expression (all P < 0.05). CONCLUSIONS: CARDS TX may participate in the immune pathogenesis of MPP through the HMGB1-TLRs/CD14-MyD88 pathway.

[1 alpha, 25-dihydroxyvitamin D3 and its analogues modulate the phagocytosis of human monocyte-derived dendritic cells].

AIM: To investigate the role of 1 alpha, 25-dihydroxyvitamin D3 (calcitriol) and its analogues tacalcitol and 24, 25(OH)2D3 on the phagocytosis of human monocyte-derived dendritic cells (MoDC). METHODS: MoDC were generated in vitro by differentiating monocytes in the presence of GM-CSF and IL-4 for 5 days. Expression of mannose receptor (MR) and Fc gamma receptors (Fc gamma Rs) by MoDC was analysed by flow cytometry. Zymosan ingestion was measured to assess the phagocytosis of MoDC. RESULTS: MoDC expressed high level of MR and Fc gamma Rs and showed the capacity of zymosan ingestion. Calcitriol and tacalcitol but no 24, 25(OH)2D3 not only upregulated the expression of MR and Fc gamma Rs on MoDC but also correspondingly enhanced their phagocytosis by increasing zymoasan ingestion. Furthermore, the upregulatory role occurred in the early stage of MoDC differentiation and was irreversible. The upregulatory role of calcitriol was dose dependent. CONCLUSION: Calcitriol and its analogue tacalcitol may play an important role in dendritic cell binding and capturing foreign antigens at the initiation of immune response.

[Effects of fumaric acid esters on differentiation of dendritic cells in vitro]

OBJECTIVE: To investigate the effect of anti-psoriatic drug fumaric acid esters (FAE) on the differentiation of dendritic cells (DC). METHODS: Dendritic cells were obtained by differentiating human monocytes in vitro. Flow cytometry was used to analyse the effect of FAE on cell surface expression of CD1a, CD14, CD40, CD80, CD86 and HLA-DR by monocyte-derived dendritic cells (MoDC). Mixed lymphocyte reaction (MLR) was made to demonstrate the influence of FAE on T cell stimulatory activity of MoDC. RESULTS: Dimethylfumarate and methylhydrogenfumarate-calcium-salt (0.01 approximate, equals 100 mg/L) inhibited MoDC differentiation as well as reducing the capacity of MoDC to stimulate lymphocytic proliferation in MLR. CONCLUSION: The mode of action of FAE in pso riasis may be mediated by inhibition of DC differentiation.

Soluble B7-H3 elevations in hospitalized children with Mycoplasma pneumoniae pneumonia.

The purpose of this study is to explore the associations between soluble B7-H3 (sB7-H3) and cytokines, clinical characteristics and laboratory findings. Thirty-two children with Mycoplasma pneumoniae pneumonia diagnosed by both positive serology and PCR were admitted to Children's Hospital affiliated to Soochow University. These children were enrolled and evaluated from May 2012 through September 2012. Soluble B7-H3 level and cytokines (tumor necrosis factor-α (TNF-α), interferon-γ, interleukin (IL)-4, IL-10) were determined by enzyme-linked immunosorbent assay technique. Meanwhile, clinical parameters including laboratory findings were obtained. Soluble B7-H3 level was significantly increased in patients with M. pneumoniae pneumonia compared with the levels of sB7-H3 in control subjects (4.94 ± 2.69 vs. 3.42 ± 1.48, ng/mL; P = 0.032). Furthermore, level of sB7-H3 was correlated with TNF-α level in plasma in patients with M. pneumoniae pneumonia (rp = 0.667; P < 0.001) as well as level of sB7-H3 in M. pneumoniae pneumonia subjects was also correlated with duration of symptoms (rp = 0.607; P < 0.001), percentage of neutrophil cells (rp = 0.657; P < 0.001), and C-reactive protein level (rs = 0.445; P = 0.011). Level of sB7-H3 was decreased after treatment (6.08 ± 3.07 vs. 3.55 ± 1.58, ng/mL; P = 0.019). Soluble B7-H3 maybe plays an important role in immunopathogenesis of M. pneumoniae pneumonia, especially for increasing TNF-α concentration and activation neutrophils.

Comparison of Recombinant Proteins from Schistosoma japonicum for Schistosomiasis Diagnosis.

The most important animal reservoirs of Schistosoma japonicum in China are bovines. Diagnosis and control of bovine schistosomiasis is critical for reducing the prevalence of the disease. We screened defined diagnostic antigens that have the potential to increase the sensitivity and specificity of serological assays and to distinguish between active and prior infections. Five recombinant proteins with the potential to be diagnostic antigens were compared to the native soluble egg antigen preparation by enzyme-linked immunosorbent assay (ELISA). We evaluated the potentials of the recombinant proteins for discriminating active from prior infections, as well as the therapeutic efficacy of the established ELISA technique.

[Effects of costimulatory pathway OX40/OX40L on the pathogenesis of allergic asthma in mice].

OBJECTIVE: Allergic asthma is thought to be mediated by CD4+ T lymphocytes producing the Th2-associated cytokines, which play a critical role in the development of the airway hyper-responsiveness and the eosinophilic inflammatory response. The costimulatory pathway CD28/B7 has been shown to play an important role in CD4+ T cell activation in allergic asthma. This study was conducted to evaluate the role of another costimulatory pathway OX40/OX40 ligand (L) in the pathogenesis of allergic asthma in BALB/c mice. METHODS: An allergic asthma model in BALB/c mice was established. Thirty-six BALB/c mice were randomly divided into three groups with 12 in each. Mice in treatment group (group B) were treated with neutralizing anti-OX40L monoclonal antibody (mAb, 300 microg per mouse) during the sensitization period. Mice in two control groups, asthma model group (group A) and IgG antibody group (group C) were treated with normal saline (NS) and control IgG respectively instead of anti-OX40L mAb. Bronchoalveolar lavage fluid (BALF) was collected from the mice of each group for counting the total number of white blood cells (including neutrophil granulocyte, lymphocyte, monocyte and eosinophil granulocyte) and the proportions of these cells. The levels of IL-4 and INF-gamma in BALF were measured by ELISA. Lungs were removed for morphological examination after HE and PAS staining, and expression of OX40 in lungs was evaluated by immunohistochemical method. RESULTS: (1) The count of total number of white blood cells in BALF (x10(6)/ml) was lower in group B than that of group A and group C (26.6 +/- 4.6 vs. 36.8 +/- 5.2 and 34.3 +/- 6.9, respectively), the difference between the treatment group (group B) and two control groups (groups A and C) was significant; The proportions of eosinophils and lymphocytes in the BALF (%) were lower in group B than those in group A and group C (eosinophils 15.1 +/- 2.6 vs. 20.0 +/- 4.1 and 19.9 +/- 3.9, respectively; lymphocytes 7.0 +/- 0.9 vs. 8.9 +/- 1.6 and 8.6 +/- 1.8, respectively), the difference between the treatment group and two control groups was significant. (2) The IL-4 level in BALF (pg/ml) was lower in group B than that in group A and group C (672 +/- 58 vs. 809.57 +/- 106.00 and 784 +/- 58, respectively), but the INF-gamma levels in BALF (pg/ml) were higher than those in group A and group C (0.86 +/- 0.09 vs. 0.69 +/- 0.15 and 0.67 +/- 0.13 respectively), and all the differences were statistically significant. (3) The expression of OX40 in the lungs of mice in group B were at a lower level than that of group A and group C, and the morphological changes of asthma were ameliorated in the mice of the treatment group. The signs of mice in treatment group were obviously ameliorated as compared to the two control groups. CONCLUSION: Blocking the costimulatory pathway by administering the neutralizing anti-OX40L mAb during the sensitization period of allergic asthma model could balance the Th1/Th2 responses, inhibit lung inflammation and ameliorate the signs of mice model of asthma.