[Analysis of the relationship between the changes of lung function and serum proinflammatory cytokines in workers occupationally exposed to toluene diisocyanate].

Objective: To analyze the correlation between the changes of lung function and serum proinflammatory cytokines in workers occupationally exposed to toluene diisocyanate (TDI), and to explore the evaluation index of respiratory toxicity of TDI. Methods: In October 2014, 61 male workers engaged in TDI synthesis process, purification process, packaging process and the above production process in a TDI factory in western China were selected as TDI exposure group; 62 male enterprise managers who were not exposed to TDI and other known allergenic chemicals were selected as control group, which were matched at the age of workers in exposure group. The questionnaire survey obtained information such as gender, length of service, age, occupational history, exposed length of service and so on. The lung function indexes [forced expiratory volume in the first second (FEV1), forced vital capacity (FVC), FEV1/FVC] and serum levels of interleukin (IL)-1 ß, IL-6, IL-8, tumor necrosis factor (TNF)-α, macrophage inflammatory factor-1 ß, monocyte chemoattractant factor-1 and vascular endothelial growth factor were measured. The urine was collected after the weekend shift, and the concentration of (TDA), the metabolite of TDI, was determined as the index of internal exposure. Spearman rank correlation was used to analyze the correlation between cytokines and lung function indexes, and multivariate linear regression was used to analyze the changes of lung function indexes and cytokines with TDI exposure concentration and time. Results: The median age (P5-P95) of the exposed group and the control group was 36.5 (24.0-51.0) and 38.0 (24.0-50.0) years, respectively. In the exposed group, the median length of service (P5-P95) was 6.94 (0.97-26.33) years, and the median concentration of TDA in urine was 15.56 (2.28-112.16) ng/ml. The three indexes of lung function, FVC, FEV1, FEV1/FVC and the levels of serum IL-8 and TNF-α were significantly lower than those in the control group (P<0.01). With the increase of exposure concentration and exposure time, the level of serum TNF-α, FVC and FEV1 decreased, and showed a good dose-effect and time-effect relationship (all Ptrend values< 0.05). Serum IL-8 and TNF-α were positively correlated with FVC, FEV1 and FEV1/FVC (all P values<0.01). Conclusion: The levels of serum inflammatory factors IL-8 and TNF-α in worker exposed to TDI are related to lung function indexes, which can be used as early evaluation indexes of respiratory toxicity induced by TDI.

Transcriptomic changes in the nasal epithelium associated with diesel engine exhaust exposure.

BACKGROUND: Diesel engine exhaust (DEE) exposure causes lung cancer, but the molecular mechanisms by which this occurs are not well understood. OBJECTIVES: To assess transcriptomic alterations in nasal epithelium of DEE-exposed factory workers to better understand the cellular and molecular effects of DEE. METHODS: Nasal epithelial brushings were obtained from 41 diesel engine factory workers exposed to relatively high levels of DEE (17.2-105.4 µg/m3), and 38 unexposed workers from factories without DEE exposure. mRNA was profiled for gene expression using Affymetrix microarrays. Linear modeling was used to identify differentially expressed genes associated with DEE exposure and interaction effects with current smoking status. Pathway enrichment among differentially expressed genes was assessed using EnrichR. Gene Set Enrichment Analysis (GSEA) was used to compare gene expression patterns between datasets. RESULTS: 225 genes had expression associated with DEE exposure after adjusting for smoking status (FDR q < 0.25) and were enriched for genes in pathways related to oxidative stress response, cell cycle pathways such as MAPK/ERK, protein modification, and transmembrane transport. Genes up-regulated in DEE-exposed individuals were enriched among the genes most up-regulated by cigarette smoking in a previously reported bronchial airway smoking dataset. We also found that the DEE signature was enriched among the genes most altered in two previous studies of the effects of acute DEE on PBMC gene expression. An exposure-response relationship was demonstrated between air levels of elemental carbon and the first principal component of the DEE signature. CONCLUSIONS: A gene expression signature was identified for workers occupationally exposed to DEE that was altered in an exposure-dependent manner and had some overlap with the effects of smoking and the effects of acute DEE exposure. This is the first study of gene expression in nasal epithelial cells of workers heavily exposed to DEE and provides new insights into the molecular alterations that occur with DEE exposure.

In vivo evaluation of anticancer efficacy of drug loaded cockle shell-derived aragonite nanoparticles.

Doxorubicin (DOX) is an effective and commonly used anthracycline anticancer drug for the treatment of osteosarcoma (OS). However, its antitumor effect is hampered by the nonspecific distribution and significant adverse effects. Nanoparticles based drug delivery systems are promising approaches to maximize the anticancer efficacy while decrease the side effects. In this study, biogenic aragonite nanoparticles (ANPs) were developed from cockle shells and loaded with DOX. An orthotopic rat OS model was induced by UMR-106 cells tibia cavity injection. The anticancer efficacy study included five groups: normal control group, OS model group, free DOX group (2 mg/kg), DOX-ANPs 1 group (2 mg of equivalent DOX/kg) and DOX-ANPs 2 group (1.5 mg of equivalent DOX/kg). This study demonstrates that the DOX-ANPs treatment groups can significantly reduce the tumor volume and increase the surviving ratio as compared to the OS model group. In addition, these two DOX-ANPs groups showed less toxicity to the normal organs compared to the free DOX group. Furthermore, DOX-ANPs 2 group showed the similar anticancer efficacy as DOX-ANPs 1 group, which suggested that DOX loaded onto the ANPs may allow the reduction of chemotherapy doses. These results highlight the promising application of ANPs derived from cockle shells as an effective drug delivery system for a successful chemotherapy against OS. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1898-1907, 2019.

In vivo evaluation of a novel nanocomposite porous 3D scaffold in a rabbit model: histological analysis.

The healing of load-bearing segmental defects in long bones is a challenge due to the complex nature of the weight that affects the bone part and due to bending, shearing, axial, and torsional forces. An innovative porous 3D scaffolds implant of CaCO3 aragonite nanocomposite derived from cockle shell was advanced for substitute bone solely for load-bearing cases. The biomechanical characteristics of such materials were designed to withstand cortical bone strength. In promoting bone growth to the implant material, an ideal surface permeability was formed by means of freeze drying and by adding copolymers to the materials. The properties of coating and copolymers supplement were also assessed for bone-implant connection resolutions. To examine the properties of the material in advanced biological system, an experimental trial in an animal model was carried out. Critical sized defect of bone was created in rabbit's radial bone to assess the material for a load-bearing application with a short and extended period assessment with histological evaluation of the incorporated implanted material to the bone of the host. Trials in animal models proved that the material has the capability of enduring load-bearing conditions for long-term use devoid of breaking or generating stress that affects the host bone. Histological examination further confirmed the improved integration of the implanted materials to the host bone with profound bone development into and also above the implanted scaffold, which was attained with negligible reaction of the tissues to a foreign implanted material.

[Evaluation of the viability of BEAS-2B cells exposed to gasoline engine exhaust with different particle sizes by air-liquid interface].

Objective: To evaluate the viability of gasoline engine exhaust (GEE) with different particle sizes on human lung cell line BEAS-2B in vitro by air-liquid interface (ALI) . Methods: GEE were collected with a Tedlar bag and their particulate matter (PM) number, surface and mass concentration in three kind of GEE (filtered automobile exhaust, non-filtered automobile exhaust and motorcycle exhaust without three-way catalytic converter) were measured by two type of particle size spectrometer including TSI-3321 and SMPS-3938. Five groups were included, which divided into blank control group, clean air group, filtered automobile exhaust group, non-filtered automobile exhaust group and motorcycle exhaust without three-way catalytic converter group. Except the blank control group, BEAS-2B cells, cultured on the surface of Transwells, were treated with clean air or GEE by ALI method at a flow rate of 25 ml/min, 37 ℃ for 60 min in vitro. CCK-8 cytotoxicity test kit was used to determine the cell relative viability of BEAS-2B cells. Results: In the filtered automobile exhaust, non-filtered automobile exhaust and motorcycle exhaust without three-way catalytic converter, high concentrations of fine particles can be detected, but the coarse particles only accounted for a small proportion, and the sequence of PM concentration was motorcycle exhaust without three-way catalytic converter group> non-filtered automobile exhaust group> filtered automobile exhaust group (P<0.001) . Compared with the clean air group, the cell relative viability in the 3 GEE-exposed groups were significantly lower (P<0.001) . Among the comparisons of GEE exposure groups with different particle size spectra, the sequence of the cell relative viability was filtered automobile exhaust group >non-filtered automobile exhaust group> motorcycle exhaust without three-way catalytic converter group (P<0.001) . When took the clean air control group as a reference, the mean of the cell relative viability in the filtered automobile exhaust group, non-filtered automobile exhaust group and motorcycle exhaust without three-way catalytic converter group, was decreased by 26.34%, 36.00% and 49.59%, respectively. Conclusion: GEE with different particle size spectra could induce different levels of toxic effects to the human lung cells BEAS-2B by ALI. After lowering the concentration of particles in the GEE and using the three-way catalytic converter could obviously improve the survival rate of lung cells.

[Association of etheno-DNA adduct and DNA methylation level among workers exposed to diesel engine exhaust].

Objective: To investigate the association between etheno-DNA adduct and the promoter of DNA methylation levels of cyclin dependent kinase inhibitor 2A (P16), Ras association domain family 1 (RASSF1A) and O-6-methylguanine-DNA methyltransferase (MGMT) in workers with occupational exposure to diesel engine exhaust (DEE). Methods: We recruited 124 diesel engine testing workers as DEE exposure group and 112 water pump operator in the same area as control group in Henan province in 2012 using cluster sampling. The demographic data were obtained by questionnaire survey; urine after work and venous blood samples were collected from each subject. The urinary etheno-DNA adducts were detected using UPLC-MS/MS, including 1,N6-etheno-2'-deoxyadenosine (εdA) and 3,N4-etheno-2'-deoxycytidine(εdC). The DNA methylation levels of P16, RASSF1A, and MGMT were evaluated using bisulfite-pyrosequencing assay. The percentage of methylation was expressed as the 5-methylcytosine (5mC) over the sum of cytosines (%5mC). Spearman correlation and multiple linear regression were applied to analyze the association between etheno-DNA adducts and DNA methylation of P16, RASSF1A, and MGMT. Results: The median (P(25)-P(75)) of urinary εdA level was 230.00 (98.04-470.91) pmol/g creatinine in DEE exposure group, and 102.10 (49.95-194.48) creatinine in control group. The level of εdA was higher in DEE exposure group than control group (P<0.001). DNA methylation levels of P16, RASSF1A and MGMT were 2.04±0.41, 2.19 (1.94-2.51), 2.22 (1.94-2.46)%5mC in exposure group, and 2.19±0.40, 2.41 (2.11-2.67), 2.44 (2.15-2.91)%5mC in control group. DNA methylation levels were lower in exposure group (P values were 0.005, 0.002 and 0.001, respectively). Spearman correlation analysis showed that DNA methylation levels of P16, RASSF1A, and MGMT were negative associated with urinary εdA level (r values were -0.155, -0.137, and -0.198, respectively, P<0.05). No significant correlation was observed between the εdC level and any measured DNA methylation levels (P>0.05) . Multiple linear regression confirmed the negative correlation between εdA and DNA methylation levels of P16, RASSF1A, and MGMT in non-smoking group (ß (95%CI) was -0.068 (-0.132--0.003), -0.082 (-0.159--0.004) and -0.048 (-0.090--0.007), P values were 0.039, 0.039 and 0.024, respectively). Moreover, εdC was negative associated with DNA methylation level of MGMT in non-smoking group (ß (95%CI) was -0.094 (-0.179--0.008), P=0.032). Conclusion: DEE exposure could induce the increased of εdA and decreased of DNA methylation levels of P16, RASSF1A and MGMT.

Further considerations in terms of the permanent impairment of the of chewing organ.

Nowadays, the drawbacks to utilizing criminal law with regards to a traumatic event, whether accidental or iatrogenic, involving the dental apparatus are still prevailing. A lesion affecting the stomatognathic apparatus can in fact integrate the offense of grievous bodily harm under Art.583 of the Italian Criminal Code (cc) as "permanent weakening of the organ" or the functions to which is appointed, i.e. the masticatory function, but also to the indwelling voice, appearance and taste. Repeatedly, jurisprudence, in the past and more recently, has set an opinion on the issue and, currently, it is possible to indicate some recognized cornerstones as deep-seated on the matter. It is a well-known fact that the organ of mastication does not represent a defined anatomical organ but, instead, it is an anatomical and functional entity consisting of a complex of different structures and not just from the dental apparatus. Moreover, any functional anatomic alteration of any single entity can have a certain weight on the other components and, therefore, on the whole organ in its entirety. Medical-legal doctrine, already in the past, supported the need for a tangible assessment of the impairment, determining whether and how much the loss of even one anatomical element could have a negative impact on the operational equilibrium of the entire stomatognathic system, whilst verifying the actual subsistence of a weakening. The loss of a dental element can take on a different meaning when such event occurs on a system already functionally deficient. In such instances, one should consider whether any pre-existing condition compromise the system, and to what extent, so that the trauma has considerable bearing on the remaining masticatory function. If the apparatus is in a position to perform even in part its function, the blunt force trauma is likely to set the aggravating circumstance.

Full-thickness skin wound healing using autologous keratinocytes and dermal fibroblasts with fibrin: bilayered versus single-layered substitute.

Split-skin grafting (SSG) is the gold standard treatment for full-thickness skin defects. For certain patients, however, an extensive skin lesion resulted in inadequacies of the donor site. Tissue engineering offers an alternative approach by using a very small portion of an individual's skin to harvest cells for propagation and biomaterials to support the cells for implantation. The objective of this study was to determine the effectiveness of autologous bilayered tissue-engineered skin (BTES) and single-layer tissue-engineered skin composed of only keratinocytes (SLTES-K) or fibroblasts (SLTES-F) as alternatives for full-thickness wound healing in a sheep model. Full-thickness skin biopsies were harvested from adult sheep. Isolated fibroblasts were cultured using medium Ham's F12: Dulbecco modified Eagle medium supplemented with 10% fetal bovine serum, whereas the keratinocytes were cultured using Define Keratinocytes Serum Free Medium. The BTES, SLTES-K, and SLTES-F were constructed using autologous fibrin as a biomaterial. Eight full-thickness wounds were created on the dorsum of the body of the sheep. On 4 wounds, polyvinyl chloride rings were used as chambers to prevent cell migration at the edge. The wounds were observed at days 7, 14, and 21. After 3 weeks of implantation, the sheep were euthanized and the skins were harvested. The excised tissues were fixed in formalin for histological examination via hematoxylin-eosin, Masson trichrome, and elastin van Gieson staining. The results showed that BTES, SLTES-K, and SLTES-F promote wound healing in nonchambered and chambered wounds, and BTES demonstrated the best healing potential. In conclusion, BTES proved to be an effective tissue-engineered construct that can promote the healing of full-thickness skin lesions. With the support of further clinical trials, this procedure could be an alternative to SSG for patients with partial- and full-thickness burns.

Peptide-binding motifs and characteristics for HLA -B*13:01 molecule.

Trichloroethylene (TCE)-induced hypersensitivity dermatitis is one of the critical occupational diseases among workers in China. Our previous studies have identified a strong linkage between the disease and the HLA-B*13:01 allele. In this study, we searched for peptides bound to the HLA-B*13:01 molecule; 57 HLA-B*13:01-bound peptides in total were identified and 54 peptides were used to calculate frequency of amino acid residues to obtain binding motifs of HLA-B*13:01 molecule. The results showed P2, P3, and P9 were the primary binding anchor positions with the dominant anchor motifs of L, Q at P2, L at P9, D at P3. HLA-B*13:01-bound peptides were identified for the first time in our research, the results of which could contribute to the human leukocyte antigen (HLA)-binding peptides database.

Brainstem auditory outcomes and correlation with neurodevelopment after perinatal asphyxia.

We used brainstem auditory-evoked responses and neurodevelopmental assessment to detect abnormalities and correlations between such responses and neurodevelopmental outcomes in 78 children (aged 4-12 years) who survived perinatal asphyxia. Twenty children had brainstem auditory-evoked response abnormalities, including increased threshold, reduced wave V amplitude, decreased V/I amplitude ratio, and prolonged I-V interval. Thirty-seven exhibited neurodevelopmental deficits, including cerebral palsy and developmental delay. The remaining 41 exhibited no deficits. Brainstem auditory-evoked response abnormalities were evident in 15 of 37 (40.5%) children with neurodevelopmental deficits, but in only 5 of 41 (12.2%) with no deficits, which differed significantly (chi(2) = 8.2, P < 0.05). The sensitivity, specificity, positive predictive value, and false-negative rate of brainstem auditory-evoked responses to reflect neurodevelopmental outcomes were 40.5%, 87.8%, 75.0%, and 59.5%, respectively. These findings suggest that in children who survive perinatal asphyxia, brainstem auditory impairment occurs more frequently in those with versus those without neurodevelopmental deficits. Brainstem auditory-evoked responses display a moderate correlation with clinically determined neurodevelopmental outcomes. Despite limitations, brainstem auditory-evoked response is valuable for assessing auditory and neurodevelopmental outcomes after perinatal asphyxia.

NMR spectroscopic evaluation of the internal environment of PLGA microspheres.

The internal environment of poly(lactide-co-glycolide) (PLGA) microspheres was characterized using 31P and 13C solid-state and solution NMR spectroscopy. Physical and chemical states of encapsulated phosphate- and histidine-containing porogen excipients were evaluated using polymers with blocked (i.e., esterified) or unblocked (free acid) end groups. Spectroscopic and gravimetric results demonstrated that the encapsulated porogen deliquesced upon hydration at 84% relative humidity to form a solution environment inside the microspheres. Dibasic phosphate porogen encapsulated in unblocked PLGA was partially titrated to the monobasic form, while in the same formulation 13C NMR showed partial protonation of the histidine imidazole. Similarly, encapsulated monobasic phosphate was partially converted to phosphoric acid. Coencapsulation of monobasic and dibasic phosphate porogens resulted in a single peak on hydration, indicating chemical exchange between discrete excipient microphases. Exogenous buffer addition differentiated external from internal, nontitratable, excipient populations. Microspheres containing dibasic phosphate porogen were hydrated with fetal calf serum, incubated at 37 degrees C, and characterized by 31P NMR through the polymer erosion phase. Within 48 h the 31P chemical shift moved over 2 ppm upfield and the line width narrowed to <60 Hz; there was little additional change through day 14. This indicated complete conversion to the monobasic phosphate form throughout the polydisperse sample and that pH remained below 4 but above the phosphoric acid p K a during matrix erosion.

Respostas cardiovasculares durante a postura sentada da Reeducação Postural Global (RPG) / Cardiovascular responses in the seated posture of the Global Postural Reeducation (GPR) method

OBJETIVO: Avaliar as respostas da freqüência cardíaca (FC), pressão arterial sistólica (PAS), diastólica (PAD), média (PAM) e duplo produto (DPr), durante a postura sentada do método de Reeducação Postural Global (RPG). MATERIAIS E MÉTODOS: Nove voluntárias saudáveis (23±2,1 anos, 56,4±7,8kg, 1,61±0,05m, 21,6±2,4kg.m2-1), inexperientes na prática do método RPG, foram submetidas a uma sessão de RPG na postura sentada, realizada em três fases: repouso pré-postura, execução da postura e recuperação pós-postura. No repouso e na recuperação, as voluntárias permaneceram sentadas por 20 minutos, sendo PA e FC verificadas a cada cinco minutos. A fase de execução da postura foi realizada em três séries e mantida por três minutos cada, com intervalo de um minuto entre elas. A verificação da PA e da FC foi realizada a cada um minuto e 30 segundos de execução da postura. RESULTADOS: Os valores de PAS, PAD, PAM e DPr foram significativamente maiores (p<0,05) do terceiro ao nono minuto da execução da postura (154±14, 107±11, 122±9mmHg e 16.478±2.802mmHg.min-1) quando comparados aos valores de repouso pré-postura (109±10, 74±7, 85±8mmHg e 9.374±1.687mmHg.min-1) e aos valores de recuperação pós-postura. Porém, estes valores retornaram aos valores de repouso nos primeiros cinco minutos de recuperação pós-postura. Durante a execução da postura, a FC não foi estatisticamente diferente da FC de repouso pré-postura. CONCLUSÕES: Elevações significativas da PAS, PAD, PAM e DPr foram observadas durante a execução da postura sentada da RPG empregada nesse estudo, mas retornaram aos valores de repouso nos primeiros cinco minutos de recuperação pós-postura.

OBJECTIVE: To evaluate heart rate (HR), systolic arterial pressure (SAP), diastolic arterial pressure (DAP), mean arterial pressure (MAP) and double product (DP) responses in the seated posture of the Global Postural Reeducation (GPR) method. METHODS: Nine healthy female volunteers (23±2.1 years; 56.4±7.8kg; 1.61±0.05m, 21.6±2.4kg/m²), without experience of the GPR, method underwent a treatment session in the seated posture. It was a three-step experiment: pre-posture resting, posture maintenance and post-posture recovery. In both the resting and the recovery step, the volunteers remained seated for 20 minutes and arterial pressure and HR were measured every five minutes. The posture maintenance step lasted for three minutes and was implemented three times with one-minute intervals between implementations. Arterial pressure and HR were measured every 1.5 minutes, while the posture was being maintained. RESULTS: The SAP, DAP, MAP and DP values were significantly greater (p<0.05) from the third to the ninth minute of maintaining the posture (154±14, 107±11, 122±9mmHg and 16,478±2,802mmHg/min) in comparison with the pre-posture resting values (109±10, 74±7, 85±8mmHg and 9,374±1,687mmHg/min) and the post-posture recovery values. However, these values returned to the resting values within the first five minutes of post-posture recovery. The HR while maintaining the posture was not statistically different from the pre-posture resting HR. CONCLUSIONS: Significant increases in SAP, DAP, MAP and DP were observed while maintaining the seated posture of the GPR method that was used in this study, but these values returned to the resting values within the first five minutes of post-posture recovery.

The comparative pharmacokinetics and pharmacodynamics of cisplatin and carboplatin in paediatric patients: a review.

Cisplatin and carboplatin are important drugs in the treatment of childhood cancers. Extensive data for carboplatin, and more limited results with cisplatin, indicate that there is significant inter-patient pharmacokinetic variability such that adaptive dosing may be necessary for the optimal use of these drugs. Glomerular filtration rate based dosing formulae have been developed for carboplatin and their use is reviewed. Further studies are needed with cisplatin in order to define the reasons for inter-patient pharmacokinetic variability and its relevance in clinical practice. Only very limited data are available on the clinical or molecular pharmacodynamics of cisplatin and carboplatin in paediatric patients. By analogy with adult studies, pharmacologically-guided platinum complex treatment should offer significant advances over empiric dosing.