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Objective: To study the clinical symptoms and the condition of skin barrier of different facial dermatitis., Methods: The patients with facial dermatitis were divided into 4 groups based on their diagnosis: hormone-dependent dermatitis (HDD) group, sensitive skin (SS) group, cosmetic intolerance (CI) group and normal control group. All patients received the evaluation on clinical symptoms with measuring scale, measurements of stratum corneum water content (SCWC), sebum, pH, trans-epidermal water loss (TEWL) and blood flow. The skin scales were collected for the measurements of the ratio of immature cornified envelope (CE) via immunofluorescence and the concentration of kallikrein 5 (KLK5) via enzyme-linked immunosorbent assay., Results: There were total 146 patients included (HDD 38, SS 53, CI 15, control group 30), most of them had their symptoms evaluated as moderate to severe. Compared with the control, the disease groups had lower SCWC and sebum (P < 0.05), higher pH, TEWL and blood flow (P < 0.05), higher ratio of immature CE (P = 0.011) and lower concentration of KLK5 (P = 0.000); while these parameters among the disease groups did not show statistical significant difference., Conclusion: There is no difference in the clinical symptoms and the condition of skin barrier between facial dermatitis with different inducements. The growth of CE may be inhibited and the concentration of KLK5 decreases in facial dermatitis.

Objective: To evaluate the efficacy of specific sublingual immunotherapy (SLIT) with Dermatophagoides farinae drops on children with allergic asthma and allergic rhinitis of the preschool and school-age groups of children and adolescents., Method: This study analyzed the efficacy of SLIT in 122 children (aged 3-14 yr) with house dust mites-induced allergic asthma and allergic rhinitis. According to the age, patients were defined as the preschool group ( ≤ 6 years old, n = 59) and school-age group (> 6 years old, n = 63). All children were treated with Dermatophagoides farinae drops for at least 2 years. Clinical observation and follow-up study were conducted during the treatment. Before and after SLIT for half a year, 1 year and 2 years, asthma symptom scores (day and night), rhinitis symptom scores and medication scores were evaluated. The adverse events were assessed during the study., Result: After SLIT for half a year, 1 year and 2 years, there were no significant differences between the preschool group (0.3 ± 0.5,0.0 ± 0.1,0.0 ± 0.0) and school-age group (0.3 ± 0.4,0.0 ± 0.1,0.0 ± 0.0) in day scores of asthma (Z value was -1.687, -0.613,0.000, all P > 0.05). There were no significant differences between the preschool group (0.2 ± 0.5,0.1 ± 0.3,0.0 ± 0.0) and school-age group (0.2 ± 0.4,0.1 ± 0.3, 0.0 ± 0.0) in night scores of asthma (Z value was -0.496, -0.486,0.000, all P > 0.05). There was no significant differences between the preschool group (1.4 ± 0.9,0.4 ± 0.5,0.1 ± 0.3) and school-age group (1.3 ± 0.9,0.5 ± 0.6,0.2 ± 0.4) in symptom scores of allergic rhinitis (Z value was -0.394, -1.166, -1.075, all P > 0.05). There were no significant differences between the preschool group (1.6 ± 0.8,0.0 ± 0.0,0.0 ± 0.0) and school-age group (1.7 ± 0.7,0.0 ± 0.0,0.0 ± 0.0) in medication scores of allergic rhinitis (Z value was -0.655,0.000,0.000, all P > 0.05). After SLIT for 2 years, most children in the preschool and school-age groups were no longer using asthma controlling medication, with no significant difference between the two groups (Z value was 0.000, P > 0.05). The overall incidence of adverse reactions was only 7%, and there was no significant difference in the incidence of adverse reactions between the two groups (χ(2) = 0.000, P > 0.05). The local adverse events were mild gastrointestinal discomfort and rash, a majority of local adverse events in the preschool group were diarrhea. No severe adverse events happened in the treatment., Conclusion: SLIT with Dermatophagoides farinae drops is safe and effective in children with allergic asthma and allergic rhinitis of the preschool and school-age groups of children and adolescents, which provides evidences for early SLIT intervention of the disease.

Aim: Dysregulation of the activity of the disintegrin/metalloproteinase ADAM10 could contribute to the development of atherosclerosis. Although a number of genetic studies have focused on the association of ADAM10 gene polymorphisms with susceptibility to diseases, no genetic association studies of ADAM10 gene variability with atherosclerotic cerebral infarction (ACI) have been conducted. The aim of this study was to analyze the potential association between ADAM10 promoter polymorphisms and ACI., Methods: The associations between rs653765 and rs514049 polymorphisms of the ADAM10 promoter and the possible risk of ACI were assessed among 347 patients with ACI and 299 matched healthy individuals in a case-control study., Results: Overall, there was a significant difference in the genotypes frequencies of rs653765 (P = 0.04) between the ACI and control subjects. In addition, the rs653765 mutated allele of ADAM10 was significantly associated with increased ADAM10 expression in patients with ACI (P = 0.032). In contrast, the allele frequency of rs514049 was not statistically associated with ACI, and the rs514049 variant A > C did not affect the expression of ADAM10 either., Conclusion: Our findings indicate a positive association between the rs653765 polymorphism of ADAM10 and ACI, as well as a negative result for rs514049. In addition, a significant increase in ADAM10 expression was observed in patients with ACI carrying the rs653765 C > T mutation. This new knowledge about ADAM10 might be clinically important and confirm a role for ADAM10 in the pathophysiology of ACI, with potentially important therapeutic implications., (© 2013 The Authors. CNS Neuroscience & Therapeutics published by John Wiley & Sons Ltd.)

Objective: Adipose tissue expressed endogenous cystathionine gamma lyase (CSE)/hydrogen sulfide (H2S) system. H2S precursor inhibited catecholamine stimulated lipolysis. Thus, we hypothesized that CSE/H2S system regulates lipolysis which contributed to the pathogenesis of insulin resistance., Methods: We treated rat adipocyte with DL-propargylglycine (PAG, a CSE inhibitor), L-cysteine (an H2S precursor) plus pyridoxial phosphate (co-enzyme) or the H2S chronic release donor GYY4137, then the glycerol level was assayed for assessing the lipolysis. Then, the effects of PAG and GYY4137 on insulin resistance in high fatty diet (HFD) induced obese mice were investigated., Results: Here, we found that PAG time-dependently increased basal or isoproterenol stimulated lipolysis. However, L-cysteine plus pyridoxial phosphate or GYY4137 significantly reduced it. PAG increased phosphorylated protein kinase A substrate, perilipin 1 and hormone sensitive lipase, but L-cysteine and GYY4137 decreased the parameters. In HFD induced obese mice, PAG increased adipose basal lipolysis, thus blunted fat mass increase, resulting in lowering insulin resistance evidenced by reduction of fasting glucose, insulin level, HOMA index, oral glucose tolerance test (OGTT) curve area and elevating the insulin tolerance test (ITT) response. GYY4137 inhibited lipolysis in vivo without increasing fat mass, but also ameliorated the insulin resistance in HFD mice., Conclusion: These results implicated that inhibition endogenous CSE/H2S system in adipocytes increased lipolysis by a protein kinase A-perilipin/hormone-sensitive lipase pathway, thus blunted fat mass increase and reduced insulin resistance in obese mice; giving H2S donor decreased lipolysis, also reduced insulin resistance induced by HFD. Our data showed that increase or decrease H2S induced opposite lipolysis, but had the same effect on insulin resistance. The paradoxical regulation may be resulted from different action of H2S on metabolic and endocrine function in adipocyte.

Objective: To establish a simple method of measuring hydrogen sulfide (H2S) in cultured living cells., Methods: Filtration membrane was stuck on the lid of cell culture plate. H2S released from cultured cells was trapped by zinc acetate to generate ZnS deposition. Then the ZnS trapped in the filtration membrane was measured by methylene blue assay and the H2S production from the living cells was counted according to the standard curve. This simple method was used to access the H2S release in HepG2 (high expression CBS and CSE) and HUVEC (low expression CSE) cell lines., Results: H2S generation in cultured HepG2 cells assayed using the present method was (859.39±19.12) nmol/(min×10(6) cells). PAG (CSE inhibitor), HA (CBS inhibitor) or the two-inhibitor (PAG+HA) treatment significantly lowered H2S release, respectively: (341.34±105.90) nmol/(min×10(6) cells), (375.05±174.50) nmol/(min×10(6) cells), and (204.47±97.14) nmol/(min×10(6) cells). The H2S production of HUVEC was (26.23±3.24) nmol/(min×10(6) cells) (about 1/30 production of HepG2 cell). Trypan blue assay showed that the cell viability was greater than 95%, suggesting that there was no cytotoxicity by using the present instrument., Conclusion: The modified instrument in cell culture plate lid was feasible for detection of hydrogen sulfide release in living cells.

In past decade, hydrogen sulfide (H₂S) as a novel gasotransmitter, covered many fields in biological and medical research. However, there is no effective, convenient and high-throughput method for determination of circulatory H₂S until now. Here, we aim to develop an easy method for measurement of circulatory H₂S by modified methylene blue method. In the present study, we added Zn²⁺ to plasma sample to deposit H₂S, HS⁻ and S²⁻, as well as plasma protein, then used NaOH to re-dissolve plasma protein. ZnS deposition was re-dissolved by the addition of N, N-dimethyl-p-phenylenediamine, and the remnant protein was deposited by trichloroacetic acid. After centrifugation, ferriammonium sulfate was added to the supernatant fluid to generate methylene blue, which was analyzed by spectrophotometer at 665 nm. Using the present method, we found that most ions including sulfate and thiosulfate did not affect detection of H₂S concentration, but albumin (physiological concentration) reduced the detection value, which suggested the binding of serum albumin and a certain amount of H₂S. The relative recovery ratio of present method is 81.9%, which implies that the method is relative accurate for the determination of H₂S concentration in plasma or serum. H₂S levels of frozen plasma samples from 65 healthy volunteers detected by the present method were (13.93 ± 4.98) µmol/L. There was no obvious difference between the detection values of fresh and frozen samples from the same SD rats. These results suggest the modified methylene blue assay is stable, sensitive, convenient and high-throughput. The method can be used to analyze the circulatory H₂S in clinical and basic research.

Objective: To investigate the expression of tumor necrosis factor-alpha (TNF-alpha) in the lung tissue of rats with lung injury induced by brain ischemia., Methods: The rat model of lung injury induced by brain ischemia was established. At 24 h, 48 h, 72 h after brain ischemia, lung tissues were harvested from each rats, the expressions of TNF-alpha mRNA and protein and its distributions in the lung tissue were measured by the methods of RT-PCR (n=8), Western blot (n=8), and immunohistochemistry (n=5), respectively., Results: The increase of TNF-alpha mRNA and protein levels were found in the lung tissues after brain ischemia, when compared with sham group (P<0.05). The morphological study with immunohistochemical staining observed TNF-alpha positive reaction in epithelial cells and some macrophages in the lung tissues after brain ischemia., Conclusion: The expression of TNF-alpha in the lung tissue could be upregulated in the rats with brain ischemia.

Fullerene [C60] included in polyethylene glycol (PEG) at a composing ratio of 1:350 w/w was examined for anticancer effects upon photodynamic therapy (PDT). Human connective tissue-derived fibrosarcoma cells HT1080 were decreased for a viability of 50% or 30%, by 3-h administration with PEG-fullerene [C60] at 50 or 100 ppm fullerene [C60] equivalent, respectively, subsequent rinsing out and irradiation with visible light (400-600 nm, 140 J/cm2: 450-fold as intense as in average outdoor), whereas the same tissue type-derived normal fibroblastic cells DUMS16 retained a viability of 93% or 85% under the same conditions. Anticancer effects were dependent on PEG-fullerene [C60] concentrations and irradiation doses, and scarcely exerted by PEG-fullerene [C60] alone, irradiation alone, or by fullerene [C60]-free PEG combined with irradiation, suggesting that the active principle may be fullerene [C60] as small as 0.0028 wt% versus the whole compound. Irradiation with PEG-fullerene [C60] occurred in intracellular DNA fragmentation according to TUNEL assay, and produced reactive oxygen species (ROS) such as hydroperoxides and peroxyl radicals or superoxide anion radicals in HT1080 cells as demonstrated by CDCFH-DA assay or nitroblue tetrazolium assay, respectively. Thus, PEG-fullerene [C60] is expected to be applied to anticancer PDT with scarce side effects on normal cells.

The development of suitable bioactive three-dimensional scaffold for the promotion of cellular proliferation and differentiation is critical in periodontal tissue engineering. In this study,porous beta-tricalcium phosphate/chitosan composite scaffolds were prepared through a freeze-drying method. These scaffolds were evaluated by analysis of microscopic structure, porosity, and cytocompatibility. The gene expression of bone sialoprotein (BSP) and cementum attachment protein (CAP) was detected with RT-PCR after human periodontal ligament cells (HPLCs) were seeded in these scaffolds. Then cell-scaffold complexes were implanted subcutaneously into athymic mice. The protein expression of alkaline phosphatase (ALP) and osteopontin (OPN) was detected in vivo. Results indicated that composite scaffolds displayed a homogeneous three-dimensional microstructure; suitable pore size (120 microm) and high porosity (91.07%). The composite scaffold showed higher proliferation rate than the pure chitosan scaffold, and up-regulated the gene expression of BSP and CAP. In vivo, HPLCs in the composite scaffold not only proliferated but also recruited vascular tissue ingrowth. The protein expression of ALP and OPN was up-regulated in the composite scaffold. Therefore, it was suggested that the composite scaffold could promote the differentiation of HPLCs towards osteoblast and cementoblast phenotypes.

Objective: To investigate the correlations of serum interleukin-18 (IL-18) level and IL-18 gene promoter polymorphisms to the development of sepsis in children., Method: Using enzyme-linked immunosorbent assay (ELISA), the authors tested the serum IL-18 level in 90 patients with sepsis and 123 normal controls, and their single nucleotide polymorphisms of the promoter region of IL-18 gene at position -607C/A and -137G/C were detected using polymerase chain reaction with sequence specific primers method and sequencing technique., Result: (1) The serum IL-18 level in sepsis groups was (196.56 +/- 157.32) pg/ml that was significantly higher than (66.16 +/- 41.63) pg/ml in normal controls (P < 0.01), the more severe the degree of sepsis was, the more significantly higher the serum IL-18 level was. The serum IL-18 level in non serious sepsis group was (152.87 +/- 114.96) pg/ml that was significantly higher than (66.16 +/- 41.63) pg/ml in normal controls, the serum IL-18 level in serious sepsis group was (191.98 +/- 169.72) pg/ml that was significantly higher than that in non serious sepsis group, and the serum IL-18 level in extremely serious sepsis patients was (323.89 +/- 159.35) pg/ml, the difference was highly significant (P = 0.000). The difference was significant among the groups with different severity of sepsis (P < 0.01). There was a negative correlation between PCIS (pediatric critical illness score) of sepsis and the serum IL-18 level (P < 0.01). (2) There were polymorphisms in IL-18 gene promoter of matched healthy children and sepsis in children. The GG genotype frequency (61.8%) of IL-18-137G/C in healthy children was the highest, followed by GC genotype (35.8%) and CC genotype (2.4%) in sequence. The G allele frequency (79.7%) was higher in IL-18-137G/C of healthy children than C allele (20.3%). The GG genotype frequency (71.1%) of IL-18-137G/C in septic children was the highest, the next were GC genotype (26.7%) and CC genotype (2.2%). The G allele frequency (84.4%) was higher in IL-18-137G/C of septic children than C allele (15.6%). The CA genotype frequency (61.0%) of IL-18-607C/A in healthy children was the highest, followed by CC genotype (26.8%) and AA genotype (12.2%). The C allele frequency (57.3%) was higher in IL-18-607C/A of healthy children than A allele (42.7%). The CA genotype frequency (76.7%) of IL-18-607C/A in septic children was the highest, followed by CC genotype (21.1%) and AA genotype (2.2%) in sequence. The C allele frequency (59.4%) was higher in IL-18-607C/A of septic children than A allele (40.6%). (3) The genotype frequency of IL-18-607 CA was 76.7% in sepsis groups that was significantly higher than 61.0% in normal controls, and the genotype frequency of -607 AA was 2.2% in sepsis groups that was significantly lower than 12.2% in normal controls, the difference was significant (P < 0.05). (4) In the order of -137CC, -137GC, -137GG, the serum IL-18 level in normal controls were as follows: (45.67 +/- 28.36) pg/ml, (53.27 +/- 37.91) pg/ml, (76.91 +/- 42.44) pg/ml, and with (140.50 +/- 60.10) pg/ml, (184.42 +/- 157.33) pg/ml, (237.02 +/- 161.76) pg/ml respectively in sepsis groups. In the order of -607AA, -607CA, -607CC, the serum IL-18 level in normal controls were: (48.80 +/- 32.11) pg/ml, (68.41 +/- 42.53) pg/ml, (70.17 +/- 43.87) pg/ml; and with (141.50 +/- 64.35) pg/ml, (151.21 +/- 121.19) pg/ml, (211.16 +/- 163.64) pg/ml respectively in sepsis groups. The difference was not significant among different groups (P > 0.05)., Conclusion: The serum IL-18 level in sepsis groups was significantly higher than that in normal controls, which was related to the severity of sepsis. It was possible that the genotype of -607CA carriers was susceptible to sepsis, which mean that the genotype of -607CA might be susceptible genotype of sepsis. However, the genotype of -607AA might play an oppose role in the risk of sepsis.

Rheumatoid arthritis (RA) is a common, systemic autoimmune disease which leads to destruction of the joint architecture and consequent disability. Although the aetiology of RA remains unknown, accumulating studies have established a strong association between RA and periodontitis (PD). Recently, anti-cyclic citrullinated peptide (anti-CCP) autoantibody and citrullinated peptide have been realized to be involved in the breaking of self-tolerance and development of autoimmune in RA. The citrullinated peptide is generated by post-translational modification (citrullination) of protein-bound arginine by peptidylarginine deiminase (PAD). Porphyromonas gingivalis(P. gingivalis), the major aetiological agent of PD and the only bacterium known to express a PAD enzyme, has been reported to be significantly associated with RA. The antibody titers to P. gingivalis are significantly increased in patients with RA and P. gingivalis antibody titers are significantly correlated with anti-CCP antibody isotypes that are specific to RA. Recent study indicates that the major synovial targets of the RA-specific anti-CCP autoantibodies are deiminated forms of the alpha- and beta- chains of fibrin. Meanwhile, it is also confirmed that bacterial PAD produced by P. gingivalis has the capacity of deiminating arginine in fibrin found in the periodontal lesion. What's more, it has been demonstrated that citrullination of HLA binding peptide causes a 100-fold increase in peptide-MHC affinity and leads to the activation CD4(+)T cells in HLA DRB1 0401 transgenic mice. Therefore, we postulate that P. gingivalis may play a crucial role in the pathogenesis of periodontitis-associated RA. P. gingivalis, which colonizes in the oral cavity, produces PAD enzyme continuously that leads to the citrullination of RA autoantigen such as fibrin in synovium joint. These PAD engendered antigens, presented in association with major histocompatibility complex (MHC) molecules by antigen-presenting cells (APC), ultimately lead to production of the anti-CCP antibody. The anti-CCP antibodies form immune complexes with citrullinated proteins, which can be bound by inflammatory cells via their Fc receptors. The roles of these immune complexes and inflammatory cells are mediated by a complex cascade involving complement activation. These mechanisms result in a release of mediators of inflammation and joint destruction ultimately leading to the onset of RA. This hypothesis reveals that oral bacterial infection may play a role in peptide citrullination which might be involved in loss of self-tolerance and development of autoimmune in RA.

The binding reaction between Vitamin B12 (B12, cyanocobalamin) and human serum albumin (HSA) was investigated by fluorescence quenching, UV-vis absorption and circular dichroism (CD) spectroscopy. Under simulative physiological conditions, fluorescence quenching data revealed that the quenching constants (Ksv) are 3.99 x 10(4), 4.33 x 10(4), 4.76 x 10(4) and 5.16 x 10(4) M(-1) at 292, 298, 304 and 310 K, respectively. The number of binding sites, n is almost constant around 1.0. On the basis of the results of fluorescence quenching the mechanism of the interaction of B12 with HSA has been found to be a dynamic quenching procedure. Thermodynamic parameters DeltaHTheta= -13.38 kJ mol(-1), DeltaSTheta=66.73 J mol(-1) K(-1) were calculated based on the binding constant. These suggested that the binding reaction was enthalpy and entropy driven, and the electrostatic interaction played major role in stabilizing the reversible complex. The binding distance r=5.5 nm between HSA and B12 was obtained according to Förster theory of energy transfer. The effect of B12 on the conformation of HSA was analyzed by synchronous fluorescence and CD spectroscopy. Synchronous spectra indicated that the polarity around the tryptophan (Trp214) residues of HSA was decreased and its hydrophobicity was increased; however, the alpha-helix content of the protein was predominant in the secondary structure but the CD spectra indicated that B12 induced minor conformational changes of HSA.

Morin [2-(2,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-1-benzopyran-4-one], a member of flavonols, is an important bioactive compound by interacting with nucleic acids, enzymes and protein. Its binding to human serum albumin was investigated by fluorescence quenching, fluorescence anisotropy, and UV-vis absorbance under the simulative physiological condition. Fluorescence quenching data show that the interaction of morin with HSA forms a non-fluorescent complex with the binding constants of 1.394 x 10(5), 1.489 x 10(5), 1.609 x 10(5) and 1.717 x 10(5)M(-1) at 292, 298, 303 and 310 K, respectively. The thermodynamics parameters, enthalpy change (DeltaH) and entropy change (DeltaS) were calculated to be 8.97 kJ mol(-1) and 129.15 J mol(-1)K(-1) via van't Hoff equation. From the spectroscopic results and thermodynamics parameters, it is observed that van der Waals and hydrogen bonds are predominant intermolecular forces when forming the complex. The distance r=4.25 nm between donor (Trp214) and accepter (morin) was estimated based on the Förster theory of non-radiative energy transfer. The red shift of UV-vis absorbance shows that morin is bound to several amino acids on the hydrophobic pocket of HSA. Moreover, the competitive probes, such as warfarin and ibuprofen (site I and II probes, respectively), reveal that the binding location of morin to HSA in the site I of the hydrophobic pocket, which corresponds to the results of UV-vis absorbance, while morin also binds other lower affinity binding sites on HSA from the fluorescence anisotropy spectroscopy.

Objective: Chronic manganese (Mn) intoxication induces syndromes resembling Parkinson disease. The clinical intervention has largely been unsuccessful. We report a 17-year follow-up study of effective treatment of occupational Mn parkinsonism with sodium para-aminosalicylic acid (PAS)., Methods: The patient, female and aged 50 at the time of treatment, was exposed to airborne Mn for 21 years (1963-1984). The patient had palpitations, hand tremor, lower limb myalgia, hypermyotonia, and a distinct festinating gait. She received 6 g PAS per day through an intravenous drip infusion for 4 days and rested for 3 days as one therapeutic course. Fifteen such courses were carried out between March and June 1987., Results: At the end of PAS treatment, her symptoms were significantly alleviated, and handwriting recovered to normal. Recent follow-up examination at age 67 years (in 2004) showed a general normal presentation in clinical, neurologic, brain magnetic resonance imaging, and handwriting examinations with a minor yet passable gait., Conclusions: This case study suggests that PAS appears to be an effective drug for treatment of severe chronic Mn poisoning with a promising prognosis.