To evaluate the chest wall shape in patients with adolescent idiopathic scoliosis (AIS) in comparison to healthy subjects and the association between the chest wall shape with the spine deformity and lung function in patients with AIS. This cross-sectional study enrolled 30 AIS patients and 20 healthy subjects aged 11-18 years old. The Cobb angle evaluation was performed in AIS patients. The chest wall shape was assessed by the photogrammetry method, using the Postural Assessment Software (PAS). We created thoracic markers shaped as angles (A) and distances (D), as follows: A2 (right acromion/xiphoid/left acromion), A4L , A7 (angle formed by the intersection of the tangent segments of the upper and lower scapulae angles), D1R/D1L [distance between the xiphoid process and the last false rib on the right (R) and left (L) sides], and D3 (distance between xiphoid process and anterior superior iliac spine).1) and D3R (r=0.47). The thoracic markers A2 and A7 were significantly higher, while the A4L and D1R/D1L were significantly reduced in the AIS group compared to the control. Moderate correlations were found between: A2 and the main and proximal thoracic Cobb angles ; D1R/D1L and the main thoracic Cobb angle (r=- 0.40); and the forced expiratory volume in the first second (FEV The photogrammetry method was able to detect chest wall changes in AIS patients, besides presenting correlation between Cobb angles and lung function. We believe that the PAS can be used as an adjunct to clinical assessment and, therefore, may prevent radiographic exposures in patients who apparently have no clinical worsening curves. Thus, we aimed to evaluate chest wall shape, using the photogrammetry method, lung function and respiratory muscle strength in patients with AIS and to compare these results with healthy subjects’ characteristics. Furthermore, we correlated chest wall shape with the Cobb method and lung function in patients with AIS.The study was conducted in a cross-sectional design from March 2014 to February 2016. We enrolled AIS patients classified as Lenke type I,Healthy adolescents, with age, weight and height range similar to AIS patients, were recruited by advertisements in local newspapers for screening and physical therapy evaluation. A single examiner, according to the Brazilian Society of Orthopedics and Traumatology recommendations,All the study participants and caregivers signed the informed assent and the consent form. The local Ethics Committee approved the present study.We evaluated 78 participants, among which 47 had been previously diagnosed with AIS and 31 were healthy individuals represented. Among the 47 AIS patients, 17 were excluded; one due to an asthma diagnosis, while the others did not belong to Lenke type I classification  and three patients did not have the main thoracic curve to the right. The excluded patients presented demographic and anthropometric characteristics similar to the included patients. Eleven participants were excluded from the control group due to postural abnormalities detected at the screening, such as presence of gibbosity in the Adam’s forward bend test, shoulders and scapular asymmetry and Thales triangle asymmetries.Thus, 30 patients and 20 healthy subjects composed the final sample. We performed the post-hoc power analysis, taking in consideration the mean (SD) of the thoracic marker A2 from the groups, with the effect size of 1.53 and the alpha error of 0.05. This calculation resulted in a sample power higher than 90%.2).After the individuals were screened, the evaluations followed the sequence presented: anthropometric assessment, respiratory evaluation and the chest wall evaluation (photogrammetry). The weight was measured to the nearest 0.1 kg, and height was measured to the nearest 0.5 cm. The body mass index was calculated by the body mass divided by the square of the body height (kg / m1), the forced vital capacity (FVC) and the FEV1/FVC ratio were analysed in absolute values and percentage of predicted values.Based on the Brazilian Thoracic Association (BTA) statement,The chest wall deformities were evaluated by a photogrammetry method, using the postural assessment software (PAS). The PAS can be accessed for free on the website (http://demotu.org/sapo/). A professional tripod was positioned half the height of the individual, where a digital camera  was positioned parallel to the ground. A plumb line marked 1 meter was placed on the ceiling of the room to calibrate the photos in the upright position. The pictures were recorded in the anterior and posterior view and left and right sides. The position of the feet was marked using an ethylene vinyl acetate (EVA) carpet. In order to mark the anatomical points of the chest region, a half-sphere of styrofoam balls of 25 mm diameter and double-sided adhesive tape were used. The anatomical points are shown in The anatomical points used were based on the PAS protocol, except point 1A1: right acromion / manubrium / left acromion;A2: right acromion / xiphoid / left acromion;A3: last false right rib / xiphoid / last false left rib;A4: angle formed between the outer point of the smallest waist circumference and its upper and lower edges;A5: point below the nipple / inferior angle of the scapula / right and left acromion;A6: C7 / acromion right and left / T3;A7: angle formed by the intersection of the tangent segments of the upper and lower scapulae angles;D1: xiphoid - last false rib on the right and left sides;D2: manubrium - last false rib on the right and left sides;D3: xiphoid - anterior superior iliac spine on the left and right sidesAll the As and Ds are illustrated in edge detector, available in the software, was used to evaluate this angle. This feature enhances the contrast of the picture, allowing better visualization of the edges in order to provide an accurate and reliable analysis. To evaluate the distances , we analysed the relationship between right and left sides .As the thoracic marker A4 was measured from the deepest point of the waist and its upper and lower edges, a tool called Descriptive analysis was expressed as mean and standard deviation and frequency and percentage. The normality of variables was investigated by Kolmogorov-Smirnov test. To differentiate the groups (control and scoliosis) regarding changes in thoracic and respiratory variables, the unpaired Student T test was used, since the variables were normally distributed. To study the correlation between chest photogrammetry and pulmonary function, respiratory muscle strength and Cobb angles, the Pearson’s correlation coefficient was calculated. The probability of alpha error was set at 5% for all analyses.The demographic and anthropometric characteristics, lung function and respiratory muscle strength of the 50 subjects evaluated and the mean Cobb angles of patients with AIS analysed in this study are shown in The chest wall evaluation by photogrammetry showed significant increase in the thoracic markers A2, A7, and significant decrease in A4L and D1R/D1L in AIS group compared to control, as shown in There were moderate positive correlation between the thoracic marker A2 and the main and proximal thoracic Cobb angles , and moderate negative correlation between the thoracic marker D1R/D1L and the main thoracic Cobb angle (r=- 0.40), as shown in 1/FVC ratio, as shown in 1 and the thoracic markers D3R  and D3L .The MIP was found to be lower in patients with AIS when compared to healthy subjects. In the assessment of pulmonary function, all variables showed decrease in patients with AIS compared to healthy subjects, with the exception of the FEVThis study consisted on assessing chest wall shape in AIS patients using the photogrammetry method. In order to quantify the chest wall alterations, we have created some angles (A) and distances (D), named thoracic markers. We noted that this method was capable to detect changes in the thoracic markers A2, A7 and D1 in AIS patients when compared to healthy subjects. Furthermore, we found that the thoracic markers A2 and D1R/D1L correlated with Cobb angles.The photogrammetry method used in our study was the postural assessment software (PAS). We suggested an interpretation for each one of them:A1, A2 and A3: tilt and rotate the ribs and sternum;A4: deviation and lateral inclination of the trunk;A5: thoracic kyphosis and/or presence of thoracic spinal deformity;A6: elevation of the shoulders;A7: uneven shoulder blades;D1 and D2: rotation of the chest wall;D3: rotation of the chest wall relative to the pelvis.We observed increase in thoracic markers A2, A7 angles and decrease in A4L angle and the D1R/D1L relationship in individuals with AIS when compared to healthy subjects. The increase of the A2 angle in AIS group may be due to vertebral rotation, in which the vertebral body turns to the convex side and the spinous process runs to the concave side. Thus, the ribs are deformed by rotating backwards and upwards on the convex side and forward on the concave side.The thoracic marker A7 was increased in AIS patients when compared to healthy individuals. According to the Brazilian Society of Orthopaedics and Traumatology, patients with AIS classified as Lenke I, with deviation of the main thoracic curve to the right, have right scapula protrusion, and the right scapula is higher than the left one in these patients; so we suggested that this marker can detect the deflection of the shoulder blades that is characteristic in these patients.The A4L angle in individuals with AIS showed significant decrease when compared to the healthy subjects. When comparing the mean values of the right (161.5±6.9) and left (144.6±9.6) A4 angles in the AIS group, we observed that the left side angle was significantly lower (p<0.001). It shows that this population presented inclination of the trunk to the left, and that this angle was able to detect this change. This result corroborates the Brazilian Society of Orthopaedics and Traumatology assessment criteria describing the inclination of the trunk to the left as one of the individualities of patients with this classification type of scoliosis.The D1R/D1L relationship was lower in the AIS group in comparison with the control group. As the ribs follow the vertebral rotation, rotating backwards and upwards on the convex side and turning forwards to the concave side,We suggested that the decrease in A5 thoracic marker could detect the presence of spinal deformity and/or thoracic kyphosis. As the patients enrolled in this study presented right gibbosity, we expected to find the A5R decreased. Analysing this marker on both sides, we noted significant decrease (p<0.001) on the right (75.2±9.1) compared to the left (82.9±9.8) side, showing that the A5 marker was capable to detect the presence of asymmetry between the sides.We also found positive correlation between the proximal thoracic Cobb angle and the A2 thoracic marker. Despite the average of the proximal thoracic Cobb angle was approximately 25º, we believe that this angle has been sufficient to change the axis of the cervical spine, and the A2 thoracic marker was able to represent the change. Probably, we found no correlation of angles and distances with thoracic lumbar curve due to the absence of a marker located in this region.1, showing that the shorter the D3 distance, the lower the FEV1. Accordingly, we suggested that the D3 thoracic marker detects rotation of the chest wall, and this rotational component may have caused distortion of the airways, causing increased resistance and reduced FEV1.We identified correlation between the thoracic markers D3 at both sides and the predicted value of FEV1 in AIS individuals were correlated to decreases in muscle strength and lung volumes when compared to healthy subjects. Studies have shown that thoracic curves have worse prognosis, because the rotation of the chest wall and its effect on the respiratory system. It has been shown negative correlation between magnitude of the scoliotic curve and the lung function.,,The decreased MIP, FVC and FEV1 were decreased in our patients with AIS, the predicted values of FVC and FEV1 in this population are within the normal range, indicating no restrictive ventilatory defect. This can be explained by the fact that the average main thoracic Cobb angle in our study was 48º, which is ranked as moderate scoliosis. Thus, even with reduction in FVC and FEV1, these values did not generate clinically important changes.Despite the FVC and FEV,Reduced MIP found in patients with AIS in our study corroborates with studies that assessed patients with thoracic scoliosis and showed that these patients have diaphragmatic dysfunction related to three-dimensional deformity of the chest, which leads to inspiratory muscle weakness.,,,,,We decided to use the PAS method for its low cost, being easy to use and capable of generating databases. Several studies have shown that the reproducibility of the PAS method is reliable.We consider that this technique should be used in conjunction with Cobb angle and not to replace it. The gold standard test for monitoring patients with scoliosis is the Cobb method, which uses plain radiography. During the growth phase, two to three radiographic studies are required, on average, by year. Radiographs are usually panoramic and have high radiation load. It is estimated that a radiograph of the lumbar spine has up to 75 times more radiation than a chest x-ray.,Although radiography is the gold standard modality in the diagnosis of scoliosis and an important tool in monitoring the disease, there must be a concern to reduce the number of radiographs of each subject to minimize costs and exposure to radioactivity.,Individuals with AIS undergoing radiographic examinations are young and, therefore, more sensitive to ionizing radiation. Ionizing radiation is known to cause cell death and genetic mutation, and it is associated with cancerous and noncancerous disease.Currently, there is concern about finding new ways to evaluate the changes of scoliosis, using non-invasive methods without the harmful effects of ionizing radiation. We believe that the PAS may be a useful method to screen the need to use the X-ray, since the physical examination may not have the sensitivity to detect subtle changes in curves.,The clinical evaluation of scoliosis is made by the Adams’ test and evaluation of the trunk asymmetry, both subjective criteria.The advantage of PAS method is that it can be used during routine assessment, in which the patient can be photographed after the anatomical landmarks are attached. Being a practical method, it allows the evaluator to measure angles and to compare them with earlier photos that can be electronically stored, becoming an objective way of assessing the deformity evolution. As an example, we can use the thoracic marker A2, which showed better correlation with the main thoracic Cobb angle and showed greater difference when compared to healthy adolescents. Moreover, in another study from our group (data in press) we observed that thoracic marker A2 showed the greatest change after scoliosis surgical treatment, proving it to be an important marker associated with deformity. Therefore, we believe that thoracic marker A2 may be a useful tool, complementing the clinical evaluation, and it may prevent at least one x-ray per year.One limitation of this study was the fact that the PAS method promotes an evaluation of the photographs in two dimensions, and scoliosis is a three-dimensional deformity of the spine. However, we found results that suggest the presence of the rotational component of the chest wall using this method. Therefore, we consider this fact did not affect the evaluation of the rotational component of the deformity.We concluded that the photogrammetry method using PAS was able to detect chest wall changes in AIS patients, besides presenting correlation between Cobb angles and lung function. Patients with AIS present reduced pulmonary function and inspiratory muscle strength. Thus, we suggested that the PAS can be used as an adjunct to clinical assessment, preventing radiologic exposures in patients who apparently have no clinical worsening curves.


The aim of the study was to evaluate the inflammatory reaction in children with pseudocroup and compare it with other laryngological diseases according to the available literature data. The study group included 51 children hospitalized because of pseudocroup. The measurements of the acute phase proteins (APP), such as C-reactive protein (CRP), alpha-1-antitrypsin (AT), alpha-1-antichymotrypsin (ACT), alpha-1-acid glycoprotein (AGP), ceruloplasmin (Cp), transferrin (Tf), alpha-2-macroglobulin (A2M), and haptoglobin (Hp) were obtained at 3 time points. The glycosylation profiles of AGP, ACT, and Tf were completed. An increased AGP level was observed in girls. The AGP glycosylation revealed the advantage of the W0 variant over the W1 variant. W1 and W2 were decreased in boys. W3 emerged in boys. The Tf concentration and T4 variant were lower compared to the control group. The A2M level was lower after treatment. The Hp and AT levels were decreased a few weeks later. The ACT glycosylation revealed a decrease of the A4 variant in boys. In conclusion, the inflammatory reaction during pseudocroup was of low intensity. The APP glycosylation suggested a chronic process. In a follow-up investigation, no normalization of the parameters was noted, but signs of persistent inflammation were observed. Subglottic laryngitis (pseudocroup) is a respiratory illness characterized by inspiratory stridor and barking cough. These symptoms result from an inflammation in the larynx at the level of the subglottic region. It occurs mainly in infants and small children between 6 and 36 months of age, more frequently in boys. Parainfluenza virus type 1 is the most common cause of pseudocroup, especially during the fall and winter epidemics when it reaches the major incidence peak –5.Different response mechanisms exist within the respiratory system. During the acute phase reaction (which is a part of the innate immune response), the basic levels of various proteins maintained by homeostatic mechanisms can change substantially. These changes are thought to contribute to host defense and other adaptive capabilities. Acute phase proteins (APP) are defined as those proteins whose serum concentrations increase or decrease by at least 25% during inflammation. Such proteins are called either positive or negative APP, respectively. The measurement of their serum levels is therefore used as a laboratory marker of the intensity of the inflammatory process. At least 40 different plasma proteins have been identified as APP. The liver is generally thought to be the main organ responsible for the secretion of all the APP into the blood as part of the systemic acute phase reaction. Several APP are widely used to monitor many diseases. Their assessment in peripheral blood represents some specificity and high sensitivity . In humaThe aim of the study was to evaluate the character and the intensity of the inflammatory reaction in children with subglottic laryngitis and compare the results with other laryngological diseases according to the available literature data.The study group included 51 children , hospitalized in the Pediatric Otolaryngology Department because of the subglottic laryngitis. Only children with symptoms requiring hospitalization, i.e., inspiratory dyspnea, stridor, and barking cough were qualified to the study group. The children with symptoms of acute lower respiratory tract infection were excluded from the study group. The results of the routine laboratory tests for the study group at the time of admission which were within normal limits are presented in The measurements of nonspecific parameters of acute phase response, such as C-reactive protein (CRP), alpha-1-antitrypsin (AT), alpha-1-antichymotrypsin (ACT), alpha-1-acid glycoprotein (AGP), ceruloplasmin (Cp), transferrin (Tf), alpha-2-macroglobulin (A2M), and haptoglobin (Hp), were obtained from each individual at 3 time points: admission (sample no. 1), after the termination of treatment (sample no. 2), and during the control visit .In the control group, which included 22 healthy children  from the kindergarten, the blood sample was obtained once during routine health checkup, and permission was obtained from the University Bioethics Committee.For the determination of the levels of APP, the Laurell rocket immunoelectrophoresis method was used and the standard solution for human serum proteins and antibodies from Dakopatts A/S (Denmark) was applied. In order to obtain the qualitative analysis of APP, the glycosylation profiles of AGP, ACT, and Tf were performed with use of the Bøg-Hansen cross-affinity immunoelectrophoresis method modified by Mackiewicz and Mackiewicz .U test was used to compare the variables between the study group and control group, and the Wilcoxon test was used for the time changes). The significance level of p < 0.05 was chosen.The distribution of all interval variables was first checked by the Shapiro-Wilk test. As all the investigated variables showed distribution distinct from normal, median and quartiles were used for the characteristics and nonparametric tests were applied . The qualitative assessment of the glycosylation profile of the AGP revealed the advantage of the W0 variant over the W1 variant. The concentration of W0 was lower in all three samples in comparison to the control group but notable only in sample no. 2 (p = 0.02). W1 was remarkably lower in all samples in comparison to the control group , while W2 was significantly higher in all 3 samples related to the control group. The appearance of the W3 variant was noted at the beginning of the disease and at the end of the treatment.The mean value of AGP was higher after the termination of treatment (sample no. 2) when compared to the day of admission (sample no. 1). During the control visit (sample no. 3), its level was lower when compared to the control group value. These differences were not statistically significant. The AGP-RC factor  was higher at the beginning of the disease and after the end of treatment, and these differences were significantly important in sample no. 2  were of statistical significance.U test value and p value are given) are presented in The results of the investigations of the APP and their glycosylation profiles for the control group and the study group at the 3 time points  consisted of 41 boys and 10 girls. The prevalence of pseudocroup was significantly more frequent in boys, with a ratio of 4.1 : 1.The study group .An increased level of AGP was observed in girls with statistical significance at admission (sample no. 1) when compared to the control group. The W1 and W2 variants were notably decreased in boys in all samples . Also, the W3 variant significantly emerged in boys in sample nos. 1 and 2  . The AGPThe alterations in ACT concentration compared to the control group and in children during pseudocroup treatment were insignificant. A slight lowering of ConA reactivity was observed for ACT in the study group, but the difference towards the control group was not significant. This indicates the absence of an intensified inflammatory process, and it can be concluded that inflammatory changes during subglottic laryngitis are not accompanied by necrotic changes, observed for example in children with chronic tonsillitis .In the first time point (sample no. 1), as well as in subsequent time points of the course of treatment, a slight decrease in Tf concentration was observed in comparison to the control group. Despite the viral etiology, inflammation activates the mechanisms that limit the availability of iron, by withdrawing Tf from the blood circulation; this is mainly the T3 variant which easily binds and releases iron and seems to correspond to apotransferrin. On the other hand, the percentage of the T1 variant, which binds iron permanently, is higher . In 2007A temporary decrease in A2M concentration was also observed in the second study compared to the control group. A2M has many diversified and complex functions, but it is primarily known by its ability to inhibit a broad spectrum of proteases without the direct blockage of the protease active site . This caDuring and because of the treatment, the inflammation in pseudocroup was relatively quickly reduced, which was followed by the significant difference between the APP intensities in the first and third time points. This applies to the initially increased AGP concentration and inflammatory W2 and W3 variants as well as the ACT-RC (ACT reactivity coefficient). The remaining APPs were only slightly altered so that the implemented treatment and recovery did not cause significant changes of their concentration .Z = −2.24; p = 0.025) and a shift of glycosylation towards a chronic process, resulting in a decrease of the W1, W2, and W3 variants.In our survey, the intensity of the inflammatory reaction during pseudocroup was higher in girls, resulting in an increased concentration of AGP and the W2 variant. In boys, differences in relation to the control group indicated a lower intensity of the inflammatory reaction , no normalization of the evaluated parameters was noted, but signs of persistence of the inflammatory process were observed


Results from mechanical properties show that the traction force is positively influenced by NaIO4, while the elongation is only affected by the PVA concentration. In summary, considering the morphological, optical and mechanical properties of the CGP/PVA films it is possible to suggest their utilization in different fields as promising packaging materials or matrices for immobilization and/or encapsulation of biomolecules.This work reports the production and characterization of biodegradable and eco-friendly films based on cashew gum polysaccharide (CGP) and polyvinyl alcohol (PVA), using the statistical design strategy. Results show that CGP/PVA films are pH stimuli-responsive, allowing their use in a magnitude of biotechnological applications. The morphological and dimensional characterization evidences a positive influence of polymers in the dimensional properties. In addition, the microstructural analysis shows that films have different morphologies depending on the content of polymers and oxidant agent. On the other hand, the thickness and light transmission values are positively influenced by CGP and PVA and negatively influenced by NaIO Plastics have become indispensable to the modern society. The reasons for their massive use are associated with their excellent properties as water and microorganism barriers, high mechanical resistance and low production cost. Their use is substantial in the package industrial sector, where they are employed in the food, pharmaceutical and cosmetic areas ,2,3. ConEven with their expressive economic advantages, the large amount of plastic wastes produced and the long time required to decompose these materials through physical, chemical and biological processes caused plastics to be associated with critical environmental issues, related to their non-biodegradability . In thisIn the last decades, biodegradable films materials based on carbohydrates, proteins and lipids have been largely studied especially due to their accessibility and lower production costs. However, biopolymer films could have some usage limitations related to their hydrophilic characteristics and barrier properties . In this4−) leads to the formation of aldehyde groups, while in hydroxyl-containing molecules, the periodate ions oxidize the hydroxyls to carboxylic acids. These reactive sites are frequently used as a chemical anchor for reactions with nucleophilic molecules, allowing the production of materials with different properties and applications [In general, the production of hybrid biopolymers is based on the introduction of reactive groups in the polymer molecules or the use of crosslinking agents. Periodate oxidation is a simple and effective method for the introduction of reactive groups in polysaccharides and hydroxyl-containing molecules. For polysaccharides, the attack by the periodate ion .The polysaccharide extraction was performed according to methodology described by Cruz et al. . Briefly4 concentrations on the biotechnological properties of the formed films, a 33-1 fractional factorial design was used (4), 10 mL of 1.0 mol L−1 phosphoric acid solution  and 15 mg of mannitol. The solutions were cast in molds and the solvent was evaporated at room temperature. The dried films were peeled from the casting mold and exhaustively washed with distilled water to completely remove unreacted materials (wash water). Then, films were dried at room temperature and stored in plastic vials. Samples of CGP/PVA films were chosen considering film uniformity and the absence of defects .The CGP/PVA films were prepared according to methodology described by Cruz et al. . In ordewas used . The filr2 = 0.995).The wash water collected as described above was used to determine the carbohydrate content according to the method described by Dubois et al.  The reac2 strips, weighted, immersed in 5 mL of different solutions and incubated at 25 °C. The swelling behavior was tested using the following solutions: (1) Ultrapure water (milli-Q); (2) 0.1 mol L−1 sodium acetate buffer pH 4.5; (3) 0.1 mol L−1 glycine buffer pH 9.5 [For the determination of the swelling index, CGP/PVA films were cut into 1 cmr pH 9.5 ,17.t was obtained using Equation (1):tW is the swollen weight of the CGP/PVA film at time t and 0W is the weight of the dried sample. The changes in the water sorption capacity of the films were monitored at regular time intervals, until reaching the swelling stabilization. Before measuring the swollen weight, films were wiped with absorbent paper to remove water on the film surface. The swelling index of each sample at time 2 strips and the three-dimensional area after 10 min of swelling was measured using a digital micrometer .Aiming to investigate the magnitude of dimensional changes during swelling, CGP/PVA films were cut into 1 cmIn order to evaluate the effect of pH on the microstructure of CGP/PVA matrixes, the swollen films were examined by the scanning electron microscopy (SEM). After swollen, the CGP/PVA films were lyophilized and the SEM analysis were performed in a JEOL JSM 6610 scanning electron microscope , using a secondary electron detector with 15 kV of acceleration. These experiments were performed in the Laboratory Laboratório Multiusuário de Microscopia de Alta Resolução (LabMic) at the Federal University of Goias, GO, Brazil. 2 were analyzed in a UV-Vis Spectrophotometer , in the wavelength ranging from 400 to750 nm, with a step size of 5 nm. The mean of transmittance values in the visible region was used as data for the 33-1 fractional factorial design.Light transmission measurements were performed according to the methodology described by Wang and Xiong , using w3-1 fractional factorial design were submitted to 10 random measurements carried out on the cross-section of the films. The thickness of the CGP/PVA films were determined as described by Cao et al. , using a−1 and an initial grid separation of 50 mm. The traction force and the elongation properties of the produced CGP/PVA films were determined in a texture analyzer , following the ASTM standard method D882-02 . Before 3-1 fractional factorial design. The models were simplified by dropping terms that have no statistical significance (p = 0.05).All analyses were performed in triplicate with repetitions, and the mean values were reported. Statistica Software  was used to perform the analysis of the data from the 3−1) and Run 6 (43.2 mg mL−1). On the other hand, films produced using 6% PVA, 1% CGP and 0.75 mol L−1 NaIO4 showed the lowest values of released carbohydrates (11.2 mg mL−1), suggesting that the CGP retention could be dependent of the PVA concentration in the formulation. In order to evaluate the retention of CGP in the CGP/PVA network, the content of unbound carbohydrates was determined in the washing waters . The hig2) and NaIO4 (X3) concentrations, the quadratic term for PVA concentration (X12) and the interaction factor for X1X22 had a positive effect on the content of released carbohydrates. This result indicates that increases in these factors will increase the amount of CGP released from the formed CGP/PVA films. On the other hand, the linear term for PVA (X1), quadratic terms for CGP (X22) and NaIO4 (X32) concentrations and the interaction factor for X1X2 negatively affected the response.Results from the multivariate analysis evidenced that all factors significantly affected the response. As can be seen in 1X2) leads to a reduction in the content of released carbohydrates. However, linear increases in PVA concentration concomitant with quadratic increases of CGP (X1X22) result in excess of unbound polysaccharides, increasing the amount of released carbohydrates.Analyzing these effects is possible to understand the relationship among the reactants in the formation of the film network. The pronounced positive effect of CGP (14.2) is a predictable result, since its excess would result in residual unbound polysaccharides, contributing to increase the released carbohydrate content. Alternatively, the expressive negative linear effect of PVA (−6.31) evidences the function of this polymer in the formation of the CGP/PVA film. The increase in PVA concentration leads to a higher CGP retention in the network. This inference is confirmed by the analysis of the interaction effect between PVA and CGP on the released carbohydrates. Generally, linear increases in both polymers (X4 concentration (2 and C3) on CGP, increasing the number of reactive groups available for chemical interactions with PVA : y denotes the content of released carbohydrates (mg mL−1), X1, X2 and X3 denote the concentration of PVA, CGP and NaIO4, respectively. Additionally, the regression analysis for released carbohydrates showed an adequate fit of experimental values to a second-order polynomial model as a function of significant factors. The mathematical model is represented by Equation (2)  revealed that the linear term of NaIO4 had the most pronounced effect on the response, negatively affecting the swelling index of the films. In addition, the quadratic term of PVA concentration and the interaction factor X1X2 also negatively affected the response. The quadratic term of CGP concentration was the only factor that positively affected the swelling index. Multivariate analysis of the results from swelling index (SI) in the acidic environment of sodium acetate buffer  indicate that the quadratic term of PVA negatively affected the swelling index (SI) in water, while the quadratic term of CGP positively affected this response. In addition, the multivariate analysis of SI in glycine buffer showed that PVA had the most pronounced effect on the swelling index, being observed as a negative effect of both linear and quadratic terms. Furthermore, the quadratic term of CGP positively affected the swelling index in the glycine buffer. It is noteworthy that the absence of the effect from NaIOa et al. . In addia et al. ,26,27,28The three-dimensional variations during the swelling of CGP/PVA films were analyzed and results are shown in 4 just affected the responses for films swollen in acetate buffer and water, negatively interfering on the response. The quadratic terms for CGP and PVA did not influence the volume values for the films swollen in acetate buffer. On the other hand, the quadratic terms for CGP (−1.46) and PVA (−5.89) negatively affected the volume values for films swollen in glycine. For the films swollen in water, the quadratic term for CGP (4.31) positively affected the films volume while the quadratic term for PVA (−3.47) had a negative effect on this response.Results from multivariate analysis showed that linear terms for PVA and CGP concentrations positively affected the values of volume for all tested solvents, whereas the content of NaIOX1, X2 and X3 denote the PVA, CGP and NaIO4 concentrations, ay, wy and yg denote the three-dimensional behavior of films swollen in acetate buffer, water and glycine buffer, respectively. The fitness was expressed by the r2 values, which indicates that more than 97% of the variability in the response can be explained by the models (Equations (3)–(5)). This suggests that the models represented accurately the data in the experimental region.The data from the volume of films swollen in the different solvents were converted into second-order polynomial equations. Consequently, the polynomial models describing the correlations between the response and the independent variables were presented as follows:Using the response surface methodology, the interaction and inter-relations of the independent variables were analyzed and the results are depicted in Scanning electronic microscopy (SEM) was performed with the aim to analyze the microstructural characteristic of the swollen CGP/PVA films . As can 4 is of major importance in the pore size and shape. In general, in the formulations produced with 1.0 mol L−1 NaIO4 there was a total absence of pores, independent of the swelling agent (films (2) and (4) from SEM micrographs evidence that the film constituents are quite important to determine the structural characteristics of the network. In addition, it is possible to infer that the concentration of NaIO4 at concentrations of 0.75 mol L−1 or 0.5 mol L−1, the SEM analysis evidenced the presence of pores differing in incidence and size. In these formulations, the polymer concentration seems to influence the degree of porosity  and CGP (X2) affected the light transmission values for both dried and wet films. However, the magnitude of factors interference was completely opposite, being observed a negative effect on the response for the dried films and a positive effect for the swollen films. Positive effects of CGP and PVA concentrations for the swollen films can be explained due to their interaction with water molecules through hydrogen bonds, increasing the amount of water inside the film and consequently decreasing the resistance to pass light throughout. On the other hand, in the dried state the chains of CGP and PVA will represent a physical barrier to the light passage, and then increasing their concentration in the film formulation will negatively affect the light transmission. The multivariate analysis results indicate that only the linear terms for PVA , but there were variations in the ratio between the area and mass of material placed in the molds for polymerization. Moreover, after polymerization, films were submitted to washes for removal of unbound polysaccharides, which may have affected the amount of material retained in the film network. In general, the thickness values of the produced CGP/PVA films were similar to those from commercial chitosan-starch films [Thickness is a physical property defined as the perpendicular distance between two surfaces. This parameter has a high importance for characterizing mechanical properties of both conventional plastics and polysaccharide-based films, affecting aspects such as uniformity, homogeneity and reproducibility ,31,32. Tch films  and lowech films  and cellch films .Results from thickness measurements shown in r = 0.46) in the range of thickness studied for CGP/PVA films. Therefore, the composition factors  have a more pronounced effect on the light transmission of the films than thickness. In fact, the action of NaIO4 in reducing the capacity of the matrix to interact with water in the medium decrease the light transmission of the films and strongly interfere in the film transparency (r = 0.99) for wet films, whereas the CGP and PVA concentration are the main factors influencing the dried film’s transparency.Thickness is a parameter that interferes in the light transmission and therefore, can alter the transparency of CGP/PVA films. Thicker films increase the absorption rate of the incident light, decreasing the transmittance detected in the spectrophotometer. Thus, we performed a Pearson’s correlation analysis between thickness values and light transmission values. Results showed a weak correlation between these parameters  on this response. The quadratic term for the CGP concentration also positively affected (55.66) the traction force, while the quadratic term for NaIO4 had a negative effect on this response (−37.13). The multivariate analysis of data from the traction force showed a significant effect of NaIOThe effects of inter-relations and interactions of the independent variables on the traction force CGP/PVA films are depicted in 4 and CGP showed an overall curvilinear profile  showed that only the linear term for PVA concentration affected the elasticity of CGP/PVA films, positively interfering (4.54) on this response. The results can be explained by the conformational structure of the PVA, which is a long linear chain being able to distend along its own axis. This ability enables the polymer matrix to extend without brake when submitted to considerable high traction forces values.Regarding to the elasticity tests , the mulComparing the mechanical properties of CGP/PVA films with literature data, it was observed that the obtained films presented similar EB values and TF values higher than those obtained for polyolefinic and starch-based films . In addiNowadays, biomaterials produced by blending natural polysaccharides with biodegradable synthetic polymers such as polyvinyl alcohol have been exhaustively tested as substitutes for petrochemical-based films ,40,41,42The high values of light transmission and tensile strength suggest that CGP/PVA films have a great potential of utilization as food packaging, especially those formulations with a higher content of PVA and CGP (Run 8 and 9), who presented tensile strength values much higher than those reported to synthetic polymers such as polyethylene and polystyrene ,45,46,47The development of biodegradable materials has also received great attention in the biomedical area, where alternative biodegradable and biocompatible scaffold materials and drug carries are extensively studied ,54. The 3-1 fractional factorial design it was possible to distinguish the effect of every component of the CGP/PVA film formulations on the chemical, physical and mechanical properties of the produced materials. First, we can determine the optimal proportions of polymers and oxidant agent to avoid waste of material. Second, the determination of the role of every component in the film formulation allows proposing the right formulation for the desired application, predicting the property of the synthesized material, what is in consonance with the consumer and market needs. Finally, considering their biodegradability and biocompatibility features, the CGP/PVA films produced in this work can be used as a renewable and eco-friendly material in substitution of the current petrochemical-based films.In summary, this work showed that using a 3


M2 for diagnostic classification models (DCMs) is under-investigated in the current literature. Specifically, the investigations of M2 for specific DCMs rather than general modeling frameworks are needed. This article aims to demonstrate the usefulness of M2 in hierarchical diagnostic classification models (HDCMs). The performance of M2 in evaluating the fit of HDCMs was investigated in the presence of four types of attribute hierarchies. Two simulation studies were conducted to examine Type I error rates and statistical power of M2 under different simulation conditions, respectively. The findings suggest acceptable Type I error rates control of M2 as well as high statistical power under the conditions of a Q-matrix misspecification and the DINA model misspecification. The data of Examination for the Certificate of Proficiency in English (ECPE) were used to empirically illustrate the suitability of M2 in practice.The performance of the limited-information statistic  M2 statistic, was recommended by researchers  fit index for M2 (RMSEA2). Similarly, the study by Henson et al.  or the most common DCMs   refers to the main effect of attribute k for item j, and k and k′ for item j. Hence, the subscript following the first comma in λj, 1, (k) or j and the examinee has mastered the first two attributes, then j, then j. Further, a statistical constraint is defined to ensure the monotonicity of the LCDM  is modified to With respect to the LCDM, j measures attributes 1 and 2 and attribute 2 is nested within attribute 1, the HDCM defines the probability of a correct response of the ith examinee for item j asAs such, if item Under the HDCM, the item parameters include only one intercept, one main effect for attribute 1 and one interaction effect for attribute 2 nested within attribute 1. Obviously, regardless of the number of attributes, there will be only one main effect in the item response function owing to the constraints of the linear hierarchies; accordingly, the number of item parameters for items measuring several attributes is also greatly reduced.j measures attributes 1, 2, and 4, the linear HDCM defines the matrix product of item j asIn this study, four fundamental hierarchical structures  under the convergent HDCM. In the divergent hierarchy, attribute 1 is the prerequisite attribute of both attributes 2 and 4, which in turn are the prerequisite attributes of attributes 3 and 5 respectively. Under the divergent HDCM, the same item would be modeled asIn the unstructured hierarchy, attribute 1 is the prerequisite attribute of all other attributes, which are independent from each other. Under the unstructured HDCM, the same item would be modeled by the same way of divergent HDCM. It should be noted that for items measuring four or five attributes, different attribute hierarchies would model the same item in more distinct ways, since more attributes lead to more main and interaction effects.2 and G2, cannot be practically feasible in DCMs. Specifically, the computations of χ2 and G2 should be based on all possible response patterns, namely, the full contingency table. However, it is required that the expected frequency in each cell should be large, i.e., usually exceeding 5, for χ2 and G2 to be effective. This indicates that only a few items and a large number of examinees are required for a test using DCMs because even a small number of items can contribute to a large number of response patterns, a situation that easily leads to the sparseness of the contingency table. As such, some approaches have been proposed to address the issue of sparseness. For instance, although the Monte Carlo resampling technique can be used to produce empirical p-values  and item response theory (IRT) framework in previous studies /2, is the number of first- and second-order residuals. Using a test of 3 items as an example, the up-to-order 2 marginal probabilities should be#, #, #)N information matrix, which is described in a recent study in detail . The Q-matrix . The C-RUM .We used the Examination for the Certificate of Proficiency in English (ECPE) data  and sparse contingency tables (J = 30/50), M2 demonstrated good control of Type I error rates.According to Simulation 1, the observed Type I error rates of n et al. , 24 itemu et al. . In theiM2 to the specification of an incorrect DINA model and the misspecification of the Q-matrix. According to Simulation 2, the M2 statistic is extremely sensitive to the misspecification of the Q-matrix regardless of sample size and attribute correlation. This finding is consistent with previous studies  in other modeling frameworks . For small sample sizes and small df, the RMSEA is often positively biased (Kenny et al., df, the model rejection rates of RMSEA were much lower for large sample sizes and large df given the same cutoff value (Chen et al., df s led to indistinguishable values and CIs of RMSEA2. More empirical investigations are needed for revealing the performance of RMSEA2 in examining the model-data fit for DCMs.Regarding the empirical illustration, the u et al. 's criterM2 statistic is of great value for examining the absolute model-data fit of HDCMs. However, given the strong sensitivity of M2 to model or Q-matrix errors, RMSEA2 is recommended to evaluate the model-data misfit.For the real-life application of DCMs, practitioners should carefully examine the relationships between attributes at the test design stage or the initial stage of data analysis. HDCMs are recommended as the modeling framework if attribute hierarchies are identified. However, the model selection decision should be based on both substantive considerations and technical solutions. Despite the fact that the model-data fit tests in DCMs are under-developed, according to our findings, the M2 in DCMs. Second, we considered only the dichotomous data in this study. Future studies regarding the application of M2 should include polytomous models. Finally, although four fundamental afttribute hierarchies were considered in our research, it is recommended that the performances of M2 and RMSEA2 be examined in DCMs with more complicated attribute relationships.Some limitations exist in the present study. First, we used a fixed test length of 20 items, which, in reality, is considered as a reasonable test length. However, future studies are encouraged to investigate the effects of test length on the performance of FC and TX contributed to the conceptualization and design of the work. FC and YL contributed to the analysis and interpretation of data. FC, YL, YC, and TX were involved in drafting and revising the manuscript. All authors approved the final manuscript submitted.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In this paper, the types, formation time, structural morphology, and influence of steel corrosion products in seawater and sea-sand concrete were studied, and the intermediate and final products of steel corrosion under different conditions were determined. The corrosion products of steel in these concrete specimens under two curing methods were studied separately by X-ray powder diffraction (XRD) and scanning electron microscopy (SEM). Due to the presence of a large amount of chloride ions in the concrete, the rust layer on the surface of a steel bar contained many intermediates, such as lepidocrocite (γ–FeOOH) and aka-ganeite (β–FeOOH). Under wet/dry cycles, with the addition and loss of moisture in concrete, various corrosion products were also dynamically converted into each other. In the specimens immersed in seawater for a long time, the intermediates of corrosion were lepidocrocite (γ–FeOOH) and aka-ganeite (β–FeOOH), which were substituted for oxygen as the new depolarizers of cathode reduction reaction, and consumed themselves to ensure smooth corrosion. With the continuous development of marine resources in China, the construction of islands is an unavoidable topic. Considering the cost of transportation and time, it is of great practical significance to use sea stones and sea-sand on reefs as concrete aggregates for materials in island engineering projects. Due to the large amount of chloride adhering to the aggregate surface of seawater and sea-sand concrete, the steel reinforcement in concrete is the first to be affected by corrosion ,3,4,5,6.For a long time, researchers have extensively studied the corrosion of steel bars in concrete caused by chloride ions, but most of the studies are focused on the corrosion rate of steel bars and the degradation of the structural properties of reinforced concrete ,10,11,12Refait et al.  found thd et al.  establisAlthough there are many studies on the effect of chloride ion on steel bars at home and abroad, in order to shorten the test period, most of them are simulated experiments of concrete simulated pore solution, accelerated corrosion of steel under current and artificial cracking. The study designs are single, and most of them investigated macro-mechanical properties, without the use of XRD, SEM, and other means from the micro perspective. However, the studies on phase composition and microstructure of rust layer in concrete are still lacking in systematic theory, which cannot accurately describe the mechanism of steel corrosion in seawater sand concrete. Therefore, the research on this aspect needs to be improved.In view of the main problems in the durability study of seawater and sea-sand concrete at present, several seawater and sea-sand concrete specimens and ordinary concrete specimens were prepared in this study, to simulate the corrosion of steel bar specimens under the conditions of continuous immersion in marine environment and dry-wet cycling, respectively, in order to study the corrosion of steel bar in the specimens. Scanning electron microscopy (SEM) was used to photograph and analyze the surface of steel bar rust layer, and the types, generation time and effect on steel bar matrix were judged according to the morphology of rust products. X-ray diffraction was used to obtain the diffraction patterns of corrosion products of different rust layers of steel bar, and the composition and internal structure of the products were obtained. According to the environment of the specimens, the main intermediate products of steel corrosion and the final oxidation products under different conditions were deduced, and the complex transformation relationship between various corrosion products was determined.Cylindrical specimens with dimensions of Φ × L = 52 mm × 320 mm were adopted to ensure that the corrosion of the embedded steel bar in all directions was identical. The cement used was P.O32.5. In the seawater and sea-sand concrete, sea stones, and sea-sand were used for aggregates, but ordinary gravel and natural medium sand were used in ordinary concrete. The fine modulus The steel bars used in the specimens were all made of the same batch of HPB235 ordinary low carbon steel with a diameter of 12 mm. The composition of the steel bars is shown in After the specimens were made, they were placed in the curing room in different groups. One group was immersed continuously in artificial seawater to simulate the underwater environment of the ocean. The other group used a wet/dry cycles device to simulate the tidal environment of the ocean. The specimens were immersed for two days and dried for two days. The water temperature was 20 ± 5 °C . The groFirst, the corroded steel bars in concrete were taken out. The corrosion layers can be divided into internal rust layer and external rust layer according to the structure. The external rust layer was loose and could be easily exfoliated. The internal rust layer with compact structure was closely connected with the reinforced bar matrix. The rust powder was grinded separately in grinding bowls until it passed through 330 mesh sieves. As shown in After the specimens were broken, the concrete was hammered into soybean-sized particles. Then, 1-2 pieces of cement paste flake particles were selected on the surface of steel bars with corrosion products on the outside of steel bars. The specimens were packed in sealed bags under vacuum to prevent the corrosion products from continuing to oxidize in contact with air. The prepared samples were analyzed by Nova NanoSEM450 field emission scanning electron microscope  . CorrosiX-ray diffraction spectra of powder samples of the inner rust layer and the outer rust layer of carbon steel rust products in the two concretes are shown in From ) is very unstable at the interface between concrete and steel matrix, and easily oxidizes into new corrosion products aka-ganeite (β–FeOOH). Misawa and others [Some scholars believe that green rust hydrochloride d others  reportedPrevious studies ,26 have The above chemical reaction not only solves the stagnation of cathodic reduction reaction caused by lack of oxygen, but also reduces the resistance of hydrated To understand the morphological characteristics of the corrosion layer of steel bars, samples A1, B1, A2 and B2 were broken and sampled, and the morphology of the corroded layer was observed by SEM. d by EDS , the corFrom Due to the long-term immersion of the specimens, it was difficult for oxygen to reach the surface of steel bars during the corrosion process. Thus, the other part of the specimens was oxidized incompletely and became black magnetite  and lepidocrocite (γ–FeOOH) dehydrate and transform into hematite  The passive film of reinforcing bar in concrete was destroyed mainly by electrochemical corrosion reaction. Under the action of oxygen and water, iron lost electrons and metal anodic dissolution occurred. The electrons obtained from oxygen and water underwent cathodic reduction reaction of depolarizer, resulting in the intermediate product (2) Due to the large amount of chloride ions present in the aggregate and mixing water of seawater sand concrete, the internal rust layer contained many transition products, including lepidocrocite (γ–FeOOH) and aka-ganeite (β–FeOOH).(3) Under wet/dry cycling conditions, steel corrosion products tended to form maghemite (γ–(4) The content of lepidocrocite (γ–FeOOH) and aka-ganeite (β–FeOOH) observed in seawater samples for a long time was relatively small. The reason is that they can replace


Tissue fibrosis, including pulmonary fibrosis, hepatic fibrosis, and cardiac fibrosis, is an important stage in the development of many diseases. It can lead to structural damage and dysfunction and even severe carcinogenesis or death. There is currently no effective method for the treatment of fibrosis. At present, the molecular mechanism of tissue fibrosis has not yet been fully elucidated, but many studies have demonstrated that it is involved in conveying the complex messages between fibroblasts and various cytokines. Sphingosine 1-phosphate (S1P) is a naturally bioactive sphingolipid. S1P and the related signaling pathways are important intracellular metabolic pathways involved in many life activities, including cell proliferation, differentiation, apoptosis, and cellular signal transduction. Increasing evidence suggests that S1P and its signaling pathways play an important role in the development of tissue fibrosis; however, the mechanisms of these effects have not yet been fully elucidated, and even the role of S1P and its signaling pathways are still controversial. This article focuses on the role of S1P and the related signaling pathways in the development of fibrosis of lung, liver, heart, and other tissues, with emphasis on the application of inhibitors of some of molecules in the pathway in clinical treatment of fibrosis diseases. Tissue fibrosis is the formation of excess fibrous connective tissue in a reparative or reactive process. It is usually observed after tissue injury in the lung, liver, heart, skin, eyes, and other organs. Some fibrotic diseases, such as idiopathic pulmonary fibrosis (IPF) and systemic sclerosis (SSc), can even lead to systemic progressive fibrosis and death .in vitro or in vivo experiments, inhibitors of this signaling pathway have been proven to effectively inhibit the development of multiple types of fibrosis, which provides a new thought for the treatment of fibrosis and the related diseases.The development of tissue fibrosis is generally sustained and progressive. At present, almost all kinds of fibrosis have no effective treatment, and the existing treatment can only improve the quality of life or prolong the life of patients to a limited extent. Therefore, it is urgent to find new precise signaling pathways or molecules participating in tissue fibrosis, and to explore new therapeutic strategies for tissue fibrosis in order to control the progressive development of the disease and even cure the disease. The S1P (sphingosine 1-phosphate) signaling pathway has become a hot spot in the research of fibrosis mechanisms in recent years because it is involved in the development of fibrosis. In many In recent years, the role of lipid in intercellular signal transduction has attracted increasing attention. The sphingomyelin (SM) signaling pathway is one of the main lipids of interest, which is involved in many activities of cells and organs, including cell survival, proliferation, differentiation, and diseases, such as cancer, infection, neurodegenerative disorders, and fibrosis.1-5. S1PRs are located in different tissues. For example, S1P1, S1P2, and S1P3 are widely expressed in multiple tissues; however, the expression of S1P4 is limited to lymphatic and hematopoietic tissues, and S1P5 is expressed in the central nervous system. Through binding to different receptors, S1P regulates many physiological or pathological processes   and S1P In recent decades, functions of S1P have been determined by several studies under normal physiological conditions. In general, there are two main functions. Firstly, S1P acts as an intracellular second messenger involved in regulating cell proliferation and inhibiting apoptosis by binding to the receptor S1PRs , whose fSphingosine 1-phosphate is widely assumed to play a protective role in the vascular system and tissues at the physiological level, while the abnormal activation of the S1P associated signaling pathway usually leads to fibrosis, cancer, and vascular dysfunction , and S1PRecent studies have shown that S1P plays a crucial role in tissue fibrosis , and thein vivo and plays an important role in life activities  and radiation exposure  can aggravate mice pulmonary fibrosis and vascular permeability, which is induced by bleomycin  and bone marrow-derived mesenchymal stem cells (BMSCs) . At the in vitro study of CCl4-induced hepatic fibrosis shows that S1P promotes the development of liver fibrosis by participating in HSCs activation and differentiation to myofibroblasts , FTY720-P, plays a role in suppressing cell proliferation and migration in cultured HSCs by inhibiting the signal pathway between S1P and PDGF , while this is not observed at lower SphK1 levels (SphK1 activity increased five times) (The expression of SphK1 in cardiac fibroblasts can affect the degeneration and fibrosis of cardiomyocytes. e times) . Howevere times) . The abo1, S1P2, and S1P3 are expressed in the heart. There are differences in the distribution of different subtypes of S1PRs in different types of cardiac cells: the main distribution of S1PR in cardiomyocytes is S1P1, while the expression of S1P2 and S1P3 are low -6 secretion depending on the Ang II-AT1, causing cardiac hypertrophy and fibrosis, and affecting the systolic function of the heart. However, this phenomenon occurs only in the proliferative fibroblasts/myocytes of the mouse heart, but not in the cardiomyocytes  and TGF-βs, which are soon released into the interstitium, and fibrosis-related signal pathways are activated, resulting in the positive feedback to promote the production and accumulation of ECM. In particular, extracellular growth factors can stimulate the paracrine effect in renal fibrosis . A studyGluteal muscle contracture (GMC) is also a chronic fibrotic disease caused by multiple etiologies, which is characterized by proliferation of fibroblasts and accumulation of collagen in the ECM of muscles . A recenSphingosine 1-phosphate and the related signaling pathway are also involved in ocular fibrosis. It can stimulate retinal pigmented epithelial cells converting into fibroblast-like cells. At the same time, S1P increases two markers of fibrosis, PAI-1 and HSP47 . Intrave1 and S1P3 have been shown to cooperate with TLR4 to promote the secretion of pro-inflammatory cytokines in endothelial and epithelial cells  induced by TGF-β and then inhibit the degradation of ECM and promote the occurrence of fibrosis  and tissue remodeling (the late stages of fibrosis) . SeveralIn addition to drug therapy, S1P is also a potential biomarker of some diseases. In a study, S1P was considered to be a potential biomarker for judging the severity of liver diseases . Howeverin vitro and in vivo studies have shown that S1P and its signaling pathways are involved in the development of multiple tissue fibrosis. Through combining with different S1PRs, S1P may play different roles in different organs and models. SphK, S1PL, and several kinds of S1P transporters also take part in the process of fibrosis by regulating the synthesis, degradation, and transfer of S1P. At the same time, S1P and the related signaling pathways compose complex signaling networks with so many signal pathways, such as inflammation, apoptosis, and autophagy, and regulate the development of fibrosis. However, the mechanism of S1P and the related signaling pathways in fibrosis have not been fully elucidated, and they have different roles in different organs and species that require further study. In addition, the intervention of S1P and the related signaling pathways are potential treatments for fibrosis-related diseases, and S1P is also expected to become an effective biomarker of the severity of fibrosis diseases in the future.At present, an effective treatment of fibrosis is still lacking. Once the fibrous tissue has formed, it is difficult to repair into normal tissue. Therefore, the studies on preventing or slowing down the development of fibrosis diseases and blocking the upstream biological processes before fibrogenesis offer better strategies for these kinds of diseases at present. A large number of EW contributed to the preparation and writing of the manuscript. MZ and XH reviewed and edited the contents of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The synergetic effect of adding multi-walled carbon nanotubes (MWCNTs) and graphene nanoplatelets (GNPs) on the thermomechanical properties and electric resistance of epoxy polymers were experimentally analyzed in this study. The total content of two employed carbon fillers were kept constant at 0.4 wt %, and seven filler ratios between two fillers (MWCNTs:GNPs), i.e., 10:0, 1:9, 3:7, 5:5, 7:3, 9:1, and 0:10, were considered in the experimental program to investigate the influences of employed nano-filler ratios on the viscoelastic and electrical properties of the studied nanocomposites. The thermomechanical properties and the sheet resistance of the nanocomposites were analyzed using a dynamic mechanical analyzer and four-point probe, respectively. Moreover, the thermogravimetric analyzer was utilized to measure the pyrolysis temperature of the nanocomposites. Experimental results show that the synergistic effect of adding two nano-fillers were clear for the improvement of the storage moduli, glass transition temperatures, and electric conductivity. Oppositely, the employment of two fillers has a slight effect on the pyrolysis temperatures of the studied nanocomposites. The composites with the MWCNT:GNP ratio of 1:9 display the most apparent enhancement of the thermomechanical properties. The improvement results from the uniform distribution and the high aspect ratio of GNPs. The addition of a small amount of MWCNTs provides more linkage in the matrix. Moreover, the specimens with the MWCNT:GNP ratio of 1:9 shows remarkable electrical properties, which result from the large contact surface areas of GNPs with each other. The employment of few MWCNTs plays an important bridging role between the layered GNPs. With the demand of electromagnetic shielding devices, conductive coatings, and antistatic materials, the conductive polymer materials have attracted more attention recently ,2,3. TheIn the past 20 years, the electrical properties ,12,13,14Since the characteristics of polymer materials are sensitive to the temperature, and the applications of carbon nanocomposites often encounter the thermal problems due to the electrical performance and surrounding environment, the knowledge of the synergistic effect on the thermomechanical properties is urgently needed. The main purpose of the study is to experimentally analyze the rheological properties of MWCNT/GNP hybrid epoxy composites, and to understand the synergistic effect of MWCNT/GNP hybrid fillers on the viscoelastic properties of the studied nanocomposites. The electrical properties and thermal stability of the MWCNT/GNP hybrid nanocomposites will be also investigated herein to evaluate the effect of adding studied hybrid fillers on the electrical properties and thermal stability. In the study, the filler ratios between two employed reinforcements will be an important variable to elucidate the influence of filler ratios on the previously mentioned properties of the nanocomposites.The matrix material employed in the present study was a two-part bisphenol A type epoxy (EPO-RT 90) supplied by Epotech Composite Corporation, Taichung, Taiwan. The mixing ratio between the epoxy resin and the hardener is 100:35. Applied Nanotechnologies, Inc., Chapel Hill, North Carolina, USA, provided the MWCNTs utilized. The purity of the studied MWCNTs is larger than 95%. 2/g. The GNPs with the designation of KNG-150 were supplied by Xiamen Knano Graphene Technology Corporation . The purity of the studied GNPs is larger than 99.5% and the specific surface area is around 40 to 60 mSeven types of specimens with different filler weight ratios were prepared for future inspections. The total content of two employed fillers are controlled at 0.4 wt %. According to a preliminary study on the room-temperature mechanical properties of MWCNT/epoxy composites with various filler contents, the nanocomposites with 0.4 wt % MWCNT have the optimal static stiffness and strength compared with those with other MWCNT contents. In the present study, the total content of two nano-fillers of the studied specimens was controlled at 0.4 wt %, and the specimens with various MWCNT:GNP ratios were prepared by replacing different contents of MWCNTs with GNPs to investigate the synergistic effect of MWCNT/GNP hybrid fillers on the thermomechanical and electrical properties. The considered MWCNT:GNP ratios in the specimen preparation are 0:10, 10:0, 1:9, 3:7, 5:5, 7:3, and 9:1. The specimens with the filler ratios of 0:10 and 10:0 represent the epoxy matrix of the specimens, which were reinforced with GNPs and MWCNTs only, respectively. In addition, the specimens of neat epoxy were also prepared for the comparison purpose.In preparing the nanocomposites specimens, the sodium dodecyl sulfate (SDS) was utilized as the surfactant to form beneficial groups on the surfaces of MWCNTs and GNPs. The SDS was added in acetone at 60 °C with mechanical stirring for 30 min. Then the nano-fillers were added in the previously mentioned solution with 10 min of mechanical stirring and 10 min of ultra-sonication to obtain the uniform distribution of the nano-fillers in the acetone solution. Next, the epoxy resin was added in the solution with 10 min mechanical stirring followed by 10 min ultra-sonication . The solution was then stirred at 100 °C until the acetone is completely volatilized. The hardener was added in the solution with ultra-sonication and low-speed mechanical stirring for 30 min. The viscous solution was vacuumed for 1 h to remove the bubbles. The de-gassed solution was then poured into a mold and vacuumed again at 100 °C for 1 h. At last, the resin was cured at 1 atm and 100 °C for 8 h to obtain the solidified nanocomposites.gT were also obtained by using the DMA inspection. The loading frequency of the DMA tests was 3 Hz, and the measured temperature ranged from 15 to 145 °C. For each type of studied composite with specific filler ratio, three samples were prepared and tested for DMA tests to examine the repeatability of obtained results.In the study, the effectiveness of treatment of carbon nano-fillers using SDS was inspected using a Fourier transform infrared spectroscopy . The morphological study was performed on the nanocomposites using a scanning electron microscope  to examine the degree of nano-filler dispersion in the epoxy matrix. To study the thermomechanical properties of the fabricated nanocomposites with various filler rations, the bending-mode dynamic mechanical analyzer  was employed to evaluate the viscous constants and rheological properties. The glass transition temperatures of the specimens dT were also obtained. The TGA tests were performed in nitrogen. The measured temperature of the TGA inspection ranged from 25 to 800 °C with the ramp rate of 10 °C/min. The TGA tests for the studied nanocomposite specimens with identical filler ratio were repeated three times to ensure the reliability of obtained data.To study the electrical properties of the nanocomposites, an Everbeing EB-6 analytical probe station  was used to measure the sheet resistances of the prospered specimens by using the four-point probe technique with a 2 mm pin interval. The studied nanocomposites with low hybrid filler concentrations (0.4 wt %) are expected to have the electrical conductivity near or below the percolation threshold of MWCNT/GNP/epoxy composites and the quantum tunneling effect is significant in the electrical conductivity ,23,24,25E′ of the studied nanocomposites with different MWNT/GNT ratios. It shows that, in the glassy region, the composites with single carbon nano-filler decrease the storage moduli of the neat epoxy. However, the employment of hybrid fillers in the epoxy improves the storage moduli significantly except for the specimens with the MWCNT:GNP ratio of 9:1. The hybrid composites with the MWCNT:GNP ratio of 1:9 display the best synergistic effect on the thermomechanical properties for the corresponding values of the storage moduli, which are higher than those with other filler ratios. The improvement results from the uniform distribution and the high aspect ratio of MWCNTs. The addition of a small amount of GNPs provides more linkage in the matrix. Surveying past literature reveals that the synergistic effect of the MWCNT/GNP hybrid fillers on the thermomechanical properties of epoxy composites is scarcely studied. The optimal filler ratios obtained by a few investigations on the room-temperature mechanical properties of the MWCNT/GNP/epoxy composites are diverse [ diverse ,10,16,21δ curves of the studied nanocomposites with various filler ratios. It shows that peaks of loss factor curves decrease with the addition of carbon nano-fillers in the pristine epoxy. Moreover, the loss factor curves tend to shift to the right when the epoxy specimens are added with the nano-fillers. The trend to the left of the loss factor curves for the hybrid nanocomposites with appropriate filler ratios is stronger than those with single filler. In gT obtained from the temperature of maximum peak of dT was obtained from the curve since the temperature corresponded to 10% weight loss. The observed pyrolysis temperatures of the specimens with various filler ratios are listed in The thermomechanical, electrical, and thermal stability properties of the epoxy composites reinforced by MWCNT and GNP hybrid fillers were experimentally studied herein to elucidate the synergistic effect of hybrid carbon nano-fillers on these properties of the studied composites. Some conclusions can be summarized as follows.(1)The results of morphological study show that the uniform dispersion of employed nano-fillers in the epoxy matrix can be obtained by applying the SDS treatment. The special hydrophobic and hydrophilic ends of SDS can avoid the agglomeration of carbon fillers and improve the dispersion effectively. Moreover, the uniform dispersion of nano-fillers in the specimen preparation ensures the repeatability of tests.(2)A strong synergistic effect of the hybrid carbon nano-fillers was found for the studied nanocomposites to improve the storage moduli and increase the glass transition temperatures.(3)The high aspect ratio of GNPs and the additional linkage provided by the few MWCNTs contribute to the significant synergistic effect on the thermomechanical properties of the composites with the MWCNT:GNP ratio of 1:9.(4)Comparing the studied composites with other filler ratios, the composites with the MWCNT:GNP ratio of 1:9 display a more intensive synergistic effect of the hybrid nano-fillers for the enhancement of electrical properties.(5)The large contact areas between the GNPs and the bridging effect provided by the MWCNTs is the main synergistic mechanism of the hybrid nano-fillers on the electrical properties of the studied composites with the MWCNT:GNP ratio of 1:9.(6)Although the addition of carbon nano-fillers can increase the pyrolysis temperatures of the studied composites, the synergistic effect of the hybrid fillers on the thermal stability of epoxy composites is not evident.


NCT03354026). Hypertensive intracerebral haemorrhage (HICH), which is characterized by rapid change, high morbidity, and mortality, is extremely dangerous. Both medical and surgical treatments lack definitive evidence and remain controversial. A prospective RCT that we have conducted has shown that the usage of the herbal medicine ICH-012 within 6 h of the event may increase the risk of haematoma enlargement and gastrointestinal bleeding. However, the volume of haematoma remains stable after 6 h. Thus, we will increase the time window to the period from 6 to 72 h after onset to evaluate the safety and efficacy of ICH-012 treating ICH  and Group C were placebo. Patients should meet all the inclusion criteria: age between 18 and 80 and diagnosis of HICH by brain CT scan between 6 and 72 h from the onset. The CT scan will be taken at four critical time points: baseline, between 6 and 72h, 24h after onset, and between 10 and 14 days after onset. The drug intervention lasts 10 days, and there is a follow-up visit taken after 90 days. The haematoma enlargement after 24 h onset as demonstrated by CT is the primary outcome. Trial registration at ClinicalTrial.gov, ID: NCT03354026, is registered on 23rd Nov. 2017. A large amount of data from high-quality RCTs is needed for the extensive clinical application of herbal medicine. The CRRICHTrial-II will evaluate the safety and effectiveness of ICH-012 in a safer time window between 6 and 72 h and investigate the possible mechanisms of action and direction of herbal medicine in the haematoma growth after HICH. Cerebrovascular disease has become the first leading cause of death in Chinese people, and haemorrhagic stroke  is one of the most important manifestations . PreventNCT01918722) in which the herbal medicine ICH-012 may increase the risk of haematoma growth and gastrointestinal bleeding without efficacy in reducing mortality or morbidity during the 6 h time window [In China, the curative effects of the traditional Chinese medicines for ICH have been demonstrated clinically, which plays an important role in reducing the mortality and disability of ICH. Herbal medicine that promotes blood circulation and reducing blood stasis was most commonly used in clinical practice –17. On tThis trial's primary aim is to evaluate the safety and efficacy of the herbal medicine ICH-012 in treating AICH within the time window of 6 to 72 h from onset. Exploring the possible mechanisms of action of the herbal medicine is also our concern.NCT03354026) involving 7 participating neurological centres. The flowchart of this research is shown in This is a prospective, randomized, double-blinded, controlled trial  The Second Teaching Hospital of Guangzhou University of Chinese Medicine, Guangzhou; (2) Lianjiang People's Hospital, Lianjiang; (3) Shenyang Second Hospital of Traditional Chinese Medicine, Shenyang; (4) The Third People's Hospital of Hubei Province, Hubei; (5) Liaocheng People's Hospital, Liaocheng; (6) Zengcheng Hospital of Traditional Chinese Medicine, Zengcheng; and (7) Cong Hua Hospital of Chinese Medicine, Conghua. Researchers from different hospitals will receive unified training and assessment before the beginning of the trial. Quality control of the trial will be undertaken by the subject team at least once a year.Men and women aged from 18 to 80 yearsAcute cerebral haemorrhage confirmed by brain CT scan within 6 to 72 h from onsetGlasgow Coma Scale ≥6Signed informed consentSecondary cerebral haemorrhage caused by brain tumour, blood diseases, cerebrovascular malformation , aneurysm, or other pathologyPatients with severe heart, liver, and renal insufficiencyIntolerance to traditional Chinese medicine (TCM) or history of allergiesPatients with severe cerebral hernia in the early onset stagePoor complianceA total of 7 hospitals across China participated in this study, including the principal responsible unit. All the participation centres are equipped with qualified neurology centres, equipment, and specialists for standardized medical care and clinical trials.α=0.05, 1-β=80%, and the primary outcome rate=20%, we used the following formula to calculate the sample size. The loss rate is supposed to be approximately 10%, and 360 patients are necessary.The enlargement percentage of the haematoma volume measured on the CT scan at the time 24 h after onset is the primary outcome of this study. We designed 3 groups  to balance and calculate the enlargement percentage of the haematoma by blinded methods. The hypothesis is that haematoma in the three groups is not different. According to a previous literature review, the rate of haematoma enlargement at 24 h after onset is likely between 14% to 20% –21. The The Chinese medical herbs used here are from the Pharmacopoeia of the People's Republic of China released on 5 June 2015. Our randomized programme was completed by the key clinical research laboratory of the Traditional Chinese Medicine Hospital of Guangdong Province. We assigned 306 cases into three groups: Groups A, B, and C in the proportion of 1:1:1. Group A, the first experimental group, used RBS, which includes 8 herbal medicines, Hirudo and Tabanus, and 6 herbals of promoting blood circulation. Group B, the second experimental group, used an herbal medicine, which includes all the herbals in Group A except Hirudo, Tabanus, and rhubarb. Group C is a placebo group with dextrin, farina, and so on. This study adopts a stratified random sampling method and intra-slice randomization on the basis of using PROC PLAN progress in SAS V9.2. In addition, the experimental measures and the control measures will be double blind. The surface of the opaque randomization envelopes will indicate the information of the test name, hospital name, and the entry sequence number of the patient. The research process of incorporating the patient, dispensing medicine depending on the random envelopes, and others will be supervised by the researchers.The first dose will be arranged within 6 to 72 h of onset, and the medicine will be taken twice a day for 10 days. The details of the treatment intervention in the three groups are shown in π/6).The first CT scan should be completed immediately after admission to identify ICH, which may be less than 6 h or between 6 and 72 h from onset. The secondary CT scan must be done after the onset time at 24 h, mainly to evaluate the haematoma growth and the oedema around the haematoma. The last CT scan can be completed within 10 to 14 days after onset, mainly to evaluate the enlargement or absorption of the haematoma. We define the amount of bleeding as the volume of the haematoma by the chef formula There are 7 secondary outcomes: (1) GCS scoring scale for consciousness assessment; (2) NIHSS scoring scale for neurological impairment assessment; (3) BI index for living ability assessment; (4) social function activity questionnaire (FAQ) to evaluate the quality of life after stroke; (5) the case fatality rate on 14 days after the onset; (6) the disability rate at the 90 day follow-up; and (7) other general questionnaires to collect information about the blood type and genotype of the patients.We record all adverse events during the experiment process in detail, and the adverse events should not be considered as a single symptom. The researchers must assign adverse events into three categories: mild, moderate and severe. Also, the possible causes and the processing method should be written on the case observation chart. If it is uncertain whether the symptom is related to the medicine, it will be recorded in detail. To judge the adverse reactions in clinical trials, we mainly follow these five principles: (1) the rationality of the time sequence of medication and adverse events; (2) adverse events reported of this herbal medicine in the past; (3) the withdrawal effects of this herbal medicine; (4) the results of the remedication after the disappearance of adverse events; and (5) considerations of other confounding factors. Lastly, the researchers will judge and decide the adverse event validation using descriptions of irrelevant, may not be relevant, probably relevant, most likely relevant, and certainly relevant.Before unblinding, only the researchers responsible for the randomized method knew the background grouping data. All the participants in this study, including the principal investigator, researchers, experimental subjects, controllers, and data analyst, will be blinded about the background data. Only in the case of severe adverse events closely related to the treatment will the blind be broken.α=0.05 (the size of the first error in hypothesis testing), 1-β=0.9 (the efficiency of the expected test), and δ=2 . In this study, the percentage change in the volume of the haematoma is measured and calculated from the baseline CT scan and the 24 h CT scan. The 7 test centres, researchers, and the CT readers are fitted with a random effect model. Also, the baseline CT scan, time course from onset to CT scan, and interventions during the research are fitted with a fixed effect model. The percentage change rate of haematoma volume will be converted to logarithmic form and normally distributed. The two treatment groups will be analysed comparatively by chi-square distribution with Bonferroni correction.The data analysis for this experiment is a comparative analysis of multiple independent sample rates, in accordance with the standard that The Data Safety Management Centre (DSM) from the Guangdong Province Hospital of Traditional Chinese Medicine is responsible for the data and safety monitoring. The data inspectors trained by the DSM will complete and submit the physical and chemical examination normal value range table to the DSM. If there is any indication of impropriety, the DSM will stop the trial and the person responsible for the trial must make a detailed description. Annual quality control meetings will be held in the duty hospital, and the researchers from all the participating hospitals must give a report about the trial progress in detail. The patients joining the trial will be registered by key information on the ClinicalTrial.gov online website.Considering the lack of studies in TCM meeting the standard of RCT, further high-quality RCTs are required . We have