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Guang-Zeng Liu,1 Yan-Gu Pan,2 Bing-Bing Li,1 Xiang-Ling Hou,3 Da-Jun Zhang11Faculty of Psychology, Research Center of Mental Health Education, Southwest University, Chongqing 400715, People’s Republic of China; 2Research Institute of Social Development, Southwestern University of Finance and Economics, Chengdu 610052, People’s Republic of China; 3School of Psychology and Cognitive Science, East China Normal University, Shanghai 200062, People’s Republic of ChinaBackground: There is some research on school climate impact on adolescent alcohol use in the Western social context; however, there is no research on school climate impact on adolescent alcohol use in China. This study aimed to explore the effect of school climate on Chinese adolescents’ alcohol use, and the moderating role of psychological suzhi between them.Methods: A total of 801 adolescents (45.8% boys, 14.96±1.66 years) completed self-reports on school climate, psychological suzhi, and alcohol use.Results: Moderation analyses revealed that both school climate and psychological suzhi significantly negatively predicted adolescents’ alcohol use, and the interaction of school climate and psychological suzhi significantly positively predicted adolescents’ alcohol use. Moreover, the effect of school climate on adolescents’ alcohol use was stronger for low psychological suzhi adolescents than high psychological suzhi adolescents.Conclusions: We can build good school climate by formulating of national level legal and regulations and good social norms, and use mature interventions or cultivation strategies to improve adolescents’ psychological suzhi in order to better play its role in protection.Keywords: school climate, psychological suzhi, alcohol use, adolescents

Clostridioides difficile infection (CDI) is an urgent public health threat with limited preventative options. In this work, we developed a messenger RNA (mRNA)-lipid nanoparticle (LNP) vaccine targeting C. difficile toxins and virulence factors. This multivalent vaccine elicited robust and long-lived systemic and mucosal antigen-specific humoral and cellular immune responses across animal models, independent of changes to the intestinal microbiota. Vaccination protected mice from lethal CDI in both primary and recurrent infection models, and inclusion of non-toxin cellular and spore antigens improved decolonization of toxigenic C. difficile from the gastrointestinal tract. Our studies demonstrate mRNA-LNP vaccine technology as a promising platform for the development of novel C. difficile therapeutics with potential for limiting acute disease and promoting bacterial decolonization.

Memory T cells are conventionally associated with durable recall responses. In our longitudinal analyses of CD4+ T cell responses to the yellow fever virus (YFV) vaccine by peptide-MHC tetramers, we unexpectedly found CD45RO-CCR7+ virus-specific CD4+ T cells that expanded shortly after vaccination and persisted months to years after immunization. Further phenotypic analyses revealed the presence of stem cell-like memory T cells within this subset. In addition, after vaccination T cells lacking known memory markers and functionally resembling genuine naive T cells were identified, referred to herein as marker-negative T (TMN) cells. Single-cell TCR sequencing detected expanded clonotypes within the TMN subset and identified TMN TCRs shared with memory and effector T cells. Longitudinal tracking of YFV-specific responses over subsequent years revealed superior stability of TMN cells, which correlated with the longevity of the overall tetramer+ population. These findings uncover additional complexity within the post-immune T cell compartment and implicate TMN cells in durable immune responses.

Aglaonema commutatum is a famous species in the Aglaonema genus, which has important ornamental and economic value. However, its chloroplast genome information and phylogenetic relationships among popular green cultivars of Aglaonema in southern China have not been reported. Herein, chloroplast genomes of one variety of A. commutatum and seven green cultivars of Aglaonema, namely, A. commutatum 'San Remo', 'Kai Sa', 'Pattaya Beauty', 'Sapphire', 'Silver Queen', 'Snow White', 'White Gem', and 'White Horse Prince', were sequenced and assembled for comparative analysis and phylogeny. These eight genomes possessed a typical quadripartite structure that consisted of a LSC region (90,799-91,486 bp), an SSC region (20,508-21,137 bp) and a pair of IR regions (26,661-26,750 bp). Each genome contained 112 different genes, comprising 79 protein-coding genes, 29 tRNA genes and 4 rRNA genes. The gene orders, GC contents, codon usage frequency, and IR/SC boundaries were highly conserved among these eight genomes. Long repeats, SSRs, SNPs and indels were analyzed among these eight genomes. Comparative analysis of 15 Aglaonema chloroplast genomes identified 7 highly variable regions, including trnH-GUG-exon1-psbA, trnS-GCU-trnG-UCC-exon1, trnY-GUA-trnE-UUC, psbC-trnS-UGA, trnF-GAA-ndhJ, ccsA-ndhD, and rps15-ycf1-D2. Reconstruction of the phylogenetic trees based on chloroplast genomes, strongly supported that Aglaonema was a sister to Anchomanes, and that the Aglaonema genus was classified into two sister clades including clade I and clade II, which corresponded to two sections, Aglaonema and Chamaecaulon, respectively. One variety and five cultivars, including A. commutatum 'San Remo', 'Kai Sa', 'Pattaya Beauty', 'Silver Queen', 'Snow White', and 'White Horse Prince', were classified into clade I; and the rest of the two cultivars, including 'Sapphire' and 'White Gem', were classified into clade II. Positive selection was observed in 34 protein-coding genes at the level of the amino acid sites among 77 chloroplast genomes of the Araceae family. Based on the highly variable regions and SSRs, 4 DNA markers were developed to differentiate the clade I and clade II in Aglaonema. In conclusion, this study provided chloroplast genomic resources for Aglaonema, which were useful for its classification and phylogeny., (© 2024. The Author(s).)

Background: Costaceae, commonly known as the spiral ginger family, consists of approximately 120 species distributed in the tropical regions of South America, Africa, and Southeast Asia, of which some species have important ornamental, medicinal and ecological values. Previous studies on the phylogenetic and taxonomic of Costaceae by using nuclear internal transcribed spacer (ITS) and chloroplast genome fragments data had low resolutions. Additionally, the structures, variations and molecular evolution of complete chloroplast genomes in Costaceae still remain unclear. Herein, a total of 13 complete chloroplast genomes of Costaceae including 8 newly sequenced and 5 from the NCBI GenBank database, representing all three distribution regions of this family, were comprehensively analyzed for comparative genomics and phylogenetic relationships., Result: The 13 complete chloroplast genomes of Costaceae possessed typical quadripartite structures with lengths from 166,360 to 168,966 bp, comprising a large single copy (LSC, 90,802 - 92,189 bp), a small single copy (SSC, 18,363 - 20,124 bp) and a pair of inverted repeats (IRs, 27,982 - 29,203 bp). These genomes coded 111 - 113 different genes, including 79 protein-coding genes, 4 rRNA genes and 28 - 30 tRNAs genes. The gene orders, gene contents, amino acid frequencies and codon usage within Costaceae were highly conservative, but several variations in intron loss, long repeats, simple sequence repeats (SSRs) and gene expansion on the IR/SC boundaries were also found among these 13 genomes. Comparative genomics within Costaceae identified five highly divergent regions including ndhF, ycf1-D2, ccsA-ndhD, rps15-ycf1-D2 and rpl16-exon2-rpl16-exon1. Five combined DNA regions (ycf1-D2 + ndhF, ccsA-ndhD + rps15-ycf1-D2, rps15-ycf1-D2 + rpl16-exon2-rpl16-exon1, ccsA-ndhD + rpl16-exon2-rpl16-exon1, and ccsA-ndhD + rps15-ycf1-D2 + rpl16-exon2-rpl16-exon1) could be used as potential markers for future phylogenetic analyses and species identification in Costaceae. Positive selection was found in eight protein-coding genes, including cemA, clpP, ndhA, ndhF, petB, psbD, rps12 and ycf1. Maximum likelihood and Bayesian phylogenetic trees using chloroplast genome sequences consistently revealed identical tree topologies with high supports between species of Costaceae. Three clades were divided within Costaceae, including the Asian clade, Costus clade and South American clade. Tapeinochilos was a sister of Hellenia, and Parahellenia was a sister to the cluster of Tapeinochilos + Hellenia with strong support in the Asian clade. The results of molecular dating showed that the crown age of Costaceae was about 30.5 Mya (95% HPD: 14.9 - 49.3 Mya), and then started to diverge into the Costus clade and Asian clade around 23.8 Mya (95% HPD: 10.1 - 41.5 Mya). The Asian clade diverged into Hellenia and Parahellenia at approximately 10.7 Mya (95% HPD: 3.5 - 25.1 Mya)., Conclusion: The complete chloroplast genomes can resolve the phylogenetic relationships of Costaceae and provide new insights into genome structures, variations and evolution. The identified DNA divergent regions would be useful for species identification and phylogenetic inference in Costaceae., (© 2024. The Author(s).)

Zingiberales includes eight families and more than 2600 species, with many species having important economic and ecological value. However, the backbone phylogenetic relationships of Zingiberales still remain controversial, as demonstrated in previous studies, and molecular dating based on chloroplast genomes has not been comprehensively studied for the whole order. Herein, 22 complete chloroplast genomes from 21 species in Zingiberales were sequenced, assembled, and analyzed. These 22 genomes displayed typical quadripartite structures, which ranged from 161,303 bp to 163,979 bp in length and contained 111-112 different genes. The genome structures, gene contents, simple sequence repeats, long repeats, and codon usage were highly conserved, with slight differences among these genomes. Further comparative analysis of the 111 complete chloroplast genomes of Zingiberales, including 22 newly sequenced ones and the remaining ones from the national center for biotechnology information (NCBI) database, identified three highly divergent regions comprising ccsA , psaC , and psaC - ndhE . Maximum likelihood and Bayesian inference phylogenetic analyses based on chloroplast genome sequences found identical topological structures and identified a strongly supported backbone of phylogenetic relationships. Cannaceae was sister to Marantaceae, forming a clade that was collectively sister to the clade of (Costaceae, Zingiberaceae) with strong support (bootstrap (BS) = 100%, and posterior probability (PP) = 0.99-1.0); Heliconiaceae was sister to the clade of (Lowiaceae, Strelitziaceae), then collectively sister to Musaceae with strong support (BS = 94-100%, and PP = 0.93-1.0); the clade of ((Cannaceae, Marantaceae), (Costaceae, Zingiberaceae)) was sister to the clade of (Musaceae, (Heliconiaceae, (Lowiaceae, Strelitziaceae))) with robust support (BS = 100%, and PP = 1.0). The results of divergence time estimation of Zingiberales indicated that the crown node of Zingiberales occurred approximately 85.0 Mya (95% highest posterior density (HPD) = 81.6-89.3 million years ago (Mya)), with major family-level lineages becoming from 46.8 to 80.5 Mya. These findings proved that chloroplast genomes could contribute to the study of phylogenetic relationships and molecular dating in Zingiberales, as well as provide potential molecular markers for further taxonomic and phylogenetic studies of Zingiberales.

In order to provide a reference for improving the physicochemical properties of starch, the study of starch polyphenol complex interaction has aroused considerable interest. As a common method of starch modification, ultrasound can make starch granules have voids and cracks, and make starch and polyphenols combine more closely. In this research, canistel seed starch was modified by ultrasonic treatment alone or combined with quercetin. The molecular structure, particle characteristics and properties of starch were evaluated. With the increase of ultrasonic temperature, the particle size of the dextrinized starch granules increased, but the addition of quercetin could protect the destruction of starch granule size by ultrasonic; X-ray diffraction and infrared spectra indicated that quercetin was bound to the surface of canistel seed starch through hydrogen bonding, and the complex and the original starch had the same crystal structure and increased crystallinity; by molecular simulation, quercetin bound inside the starch molecular helix preserved the crystalline helical configuration of starch to some extent and inhibited the complete unhelicalization of starch molecules. Meanwhile, hydrogen bonding was the main driving force for the binding of starch molecules to quercetin, and van der Waals interactions also promoted the binding of both. In the physicochemical properties, as the temperature increased after the combination of ultrasonic modified starch combined with quercetin, the solubility, swelling force and apparent viscosity of the compound increased significantly, and it has higher stability and shear resistance., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Dystonia is a genetically and phenotypically heterogeneous disorder that occurs in isolation (isolated dystonia) or in combination with other movement disorders. To determine the genetic spectrum in isolated dystonia, we enrolled 88 patients with isolated dystonia for whole-exome sequencing (WES). Seventeen mutations, including nine novel ones, were identified in 19 of the 88 patients, providing a 21.59% positive molecular diagnostic rate. Eleven distinct genes were involved, of which TOR1A and THAP1 accounted for 47.37% (9/19) of the positive cases. A novel missense variant, p.S225R in TOR1A, was found in a patient with adolescence-onset generalized dystonia. Cellular experiments revealed that p.S255R results in the abnormal aggregation of Torsin-1A encoding by TOR1A. In addition, we reviewed the clinical and genetic features of the isolated dystonia patients carrying TOR1A, THAP1, ANO3, and GNAL mutations in the Chinese population. Our results expand the genetic spectrum and clinical profiles of patients with isolated dystonia and demonstrate WES as an effective strategy for the molecular diagnosis of isolated dystonia., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The baseline composition of T cells directly affects later response to pathogens, but the complexity of precursor states remains poorly defined. Here, we examined the baseline state of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific T cells in unexposed individuals. SARS-CoV-2-specific CD4 + T cells were identified in prepandemic blood samples by major histocompatibility complex (MHC) class II tetramer staining and enrichment. Our data revealed a substantial number of SARS-CoV-2-specific T cells that expressed memory phenotype markers. Integrated phenotypic analyses demonstrated diverse preexisting memory states that included cells with distinct polarization features and trafficking potential to barrier tissues. T cell clones generated from tetramer-labeled cells cross-reacted with antigens from commensal bacteria in the skin and gastrointestinal tract. Direct ex vivo tetramer staining for one spike-specific population showed a similar level of cross-reactivity to sequences from endemic coronavirus and commensal bacteria. These data highlight the complexity of precursor T cell repertoire and implicate noninfectious exposures to common microbes as a key factor that shapes human preexisting immunity to SARS-CoV-2.

The flat-joint model, which constructs round particles as polygons, can suppress rotation after breakage between particles and simulate more larger compression and tension ratios than the linear parallel-bond model. The flat-joint contact model was chosen for this study to calibrate the rock for 3D experiments. In the unit experiments, the triaxial unit was loaded with flexible boundaries, and the influence of each microscopic parameter on the significance magnitude of the macroscopic parameters (modulus of elasticity E , Poisson's ratio ν , uniaxial compressive strength UCS , crack initiation strength σ ci , internal friction angle φ and uniaxial tensile strength TS ) was analysed by ANOVA (Analysis of Variance) in an orthogonal experimental design. Among them, E ƒ , k ƒ has a significant effect on E ; C ƒ and k ƒ have a significant effect on ν ; C ƒ , σ ƒ and k ƒ have a significant effect on UCS ; C ƒ ; σ ƒ and E ƒ have a significant effect on TS; R sd has a significant effect on σ ci ; and φ f , E ƒ , k ƒ , μ ƒ , and σ ƒ have a significant effect on φ . Regressions were then carried out to establish the equations for calculating the macroscopic parameters of the rock material so that the three-dimensional microscopic parameters of the PFC can be quantitatively analysed and calculated. The correctness of the establishment of the macroscopic equations was verified by comparing the numerical and damage patterns of uniaxial compression, Brazilian splitting, and triaxial experiments with those of numerical simulation units in the chamber.

Objective: To explore the effects of porcine acellular dermal matrix (ADM) combined with human epidermal stem cells (ESCs) on wound healing of full-thickness skin defect in nude mice. Methods: The morphology of porcine ADM was analyzed by photograph of digital camera, the cell residues in porcine ADM were observed by hematoxylin-eosin (HE) staining, the surface structure of porcine ADM was observed by scanning electron microscope, the secondary structure of porcine ADM was analyzed by infrared spectrometer, the porcine ADM particle size was analyzed by dynamic light scattering particle size analyzer, and the porcine ADM potential was analyzed by nano-particle size potentiometer. The morphology of porcine ADM was observed by inverted fluorescence microscope when it was placed in culture medium for 30 min, 1 d, and 5 d ( n =2). The porcine ADM was divided into 5 min group, 10 min group, 20 min group, 30 min group, 60 min group, and 120 min group according to the random number table (the same grouping method below) in static state at normal temperature for the corresponding time to calculate the water absorption by weighing method ( n =3). Swiss white mouse embryonic fibroblasts (Fbs) were divided into blank control group (culture medium only), and 50.0 g/L ADM extract group, 37.5 g/L ADM extract group, 25.0 g/L ADM extract group, 12.5 g/L ADM extract group, and 6.5 g/L ADM extract group which were added with the corresponding final concentrations of ADM extract respectively. At post culture hour (PCH) 24, 48, and 72, the cell survival rate was detected by cell counting kit 8 and the cytotoxicity was graded ( n =5). The erythrocytes of a 6-week-old male Sprague-Dawley male rat were divided into normal saline group, ultra-pure water group, and 5 mg/mL ADM extract group, 10 mg/mL ADM extract group, and 15 mg/mL ADM extract group which were treated with the corresponding final concentrations of porcine ADM extract respectively. After reaction for 3 h, the absorbance value of hemoglobin was detected by microplate reader to represent the blood compatibility of porcine ADM ( n =3). ESCs were isolated and cultured from the discarded prepuce of a 6-year-old healthy boy who was treated in the Department of Urology of the First Affiliated Hospital of Army Medical University (the Third Military Medical University) in July 2020, and then identified by flow cytometry. The porcine ADM particles of composite ESC (hereinafter referred to as ESC/ADM) were constructed by mixed culture. After 3 days of culture, the composite effect of ESC/ADM was observed by HE staining and laser scanning confocal microscope. Thirty-six 7-8-week-old male non-thymic nude mice were divided into phosphate buffer solution (PBS) alone group, ADM alone group, ESC alone group, and ESC/ADM group, with 9 mice in each group, and the wound model of full-thickness skin defect was established. Immediately after injury, the wounds were treated with the corresponding reagents at one time. On post injury day (PID) 1, 7, 11, and 15, the wound healing was observed and the wound healing rate was counted ( n =3). On PID 7, the epithelialization of wounds was observed by HE staining and the length of un-epithelialized wound was measured (with this and the following sample numbers of 4). On PID 11, the dermal area and collagen deposition of wounds were observed by Masson staining and the dermal area of wound section was calculated, the number of cells expressing CD49f, a specific marker of ESC, was calculated with immunofluorescence staining, the mRNA expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in ESC after wound transplantation was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction. Data were statistically analyzed with independent sample t test, one-way analysis of variance, analysis of variance for repeated measurement, and least significant difference t test. Results: The porcine ADM was white particles and composed of reticular structure, with no cells inside, disordered structure, and rough surface. The absorption peak of porcine ADM appeared at the wave numbers of 1 659, 1 549, and 1 239 cm -1 , respectively. The main particle size distribution of porcine ADM in solution was 500 to 700 nm, with negative charge on the surface. The morphology of porcine ADM in static state at 30 min and on 1 and 5 d was relatively stable. The water absorption of porcine ADM remained relatively high level in static state from 30 min to 120 min. The cytotoxicity of mouse embryonic Fbs in 6.5 g/L ADM extract group, 12.5 g/L ADM extract group, and 25.0 g/L ADM extract group was grade 1 at PCH 24, and the cytotoxicity of the other groups was 0 grade at each time point. After reaction for 3 h, the absorbance value of hemoglobin of erythrocytes in ultra-pure water group was significantly higher than the values in normal saline group and 15 mg/mL ADM extract group (with t values of 8.14 and 7.96, respectively, P <0.01). After 3 days of culture, the cells of the fourth passage showed pebble-like morphology, with low expression of CD71 and high expression of CD49f, which were identified as ESCs. There was ESC attachment and growth on porcine ADM particles. On PID 1, the wound sizes of nude mice were almost the same in PBS alone group, ADM alone group, ESC alone group, and ESC/ADM group. On PID 7, 11, and 15, the wound contraction of nude mice in each group was observed, especially in ADM alone group, ESC alone group, and ESC/ADM group. On PID 7, the wound healing rates of nude mice in ESC alone group and ESC/ADM group were significantly higher than the rate in PBS alone group (with t values of 2.83 and 4.72 respectively, P <0.05 or P <0.01). On PID 11, the wound healing rate of nude mice in ESC/ADM group was significantly higher than that in PBS alone group ( t =4.86, P <0.01). On PID 15, the wound healing rates of nude mice in ADM alone group, ESC alone group, and ESC/ADM group were significantly higher than the rate in PBS alone group (with t values of 2.71, 2.90, and 3.23 respectively, P <0.05). On PID 7, the length of un-epithelialized wound of nude mice in ADM alone group, ESC alone group, and ESC/ADM group was (816±85), (635±66), and (163±32) μm, respectively, which were significantly shorter than (1 199±43) μm in PBS alone group (with t values of 5.69, 10.19, and 27.54 respectively, P <0.01). On PID 11, the dermal areas of wound section of nude mice in ADM alone group, ESC alone group, and ESC/ADM group were significantly larger than the area in PBS alone group (with t values of 27.14, 5.29, and 15.90 respectively, P <0.01); the collagen production of nude mice in ADM alone group and ESC/ADM group was more obvious than that in PBS alone group, and the collagen production of nude mice in ESC alone group and PBS alone group was similar. On PID 11, in the wounds of nude mice in ESC alone group and ESC/ADM group, the cells with positive expression of CD49f were respectively 135±7 and 185±15, and the mRNA expressions of GAPDH were positive; while there were no expressions of CD49f nor mRNA of GAPDH in the wounds of nude mice in PBS alone group and ADM alone group. Conclusions: ESC/ADM particles can promote the wound healing of full-thickness skin defects in nude mice, which may be related to the improved survival rate of ESCs after transplantation and the promotion of dermal structure rearrangement and angiogenesis by ADM.

The development of peptide-major histocompatibility complex tetramers has enabled direct characterization and enumeration of antigen-specific T cells. However, the weaker interaction between class II tetramers and CD4 + T cells increases the challenge of using tetramers to analyze CD4 + T cell responses. Here, we provide an optimized class II tetramer staining protocol with a magnetic-bead enrichment strategy for the detection and functional analyses of human antigen-specific CD4 + T cells. This approach enables direct sampling of lymphocytes that recognize specific peptide-MHC complexes, including rare pathogen-specific CD4 + T cells from unexposed individuals., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Sensory ataxic neuropathy, dysarthria, and ophthalmoparesis (SANDO) is a rare mitochondrial disorder associated with mutations in the POLG gene, which encodes the DNA polymerase gamma catalytic subunit. A few POLG-related SANDO cases have been reported, but the genotype-phenotype correlation remains unclear. Here, we report a patient with SANDO carrying two novel missense variants (c.2543G>C, p.G848A and c.452 T>C, p.L151P) in POLG. We also reviewed previously reported cases to systematically evaluate the clinical and genetic features of POLG-related SANDO. A total of 35 distinct variants in the coding region of POLG were identified in 63 patients with SANDO. The most frequent variant was the p.A467T variant, followed by the p.W748S variant. The clinical spectrum of SANDO is heterogeneous. No clear correlation has been observed between the mutation types and clinical phenotypes. Our findings expand the mutational spectrum of POLG and contribute to clinical management and genetic counseling for POLG-related SANDO., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

We examined how baseline CD4 + T cell repertoire and precursor states impact responses to pathogen infection in humans using primary immunization with yellow fever virus (YFV) vaccine. YFV-specific T cells in unexposed individuals were identified by peptide-MHC tetramer staining and tracked pre- and post-vaccination by tetramers and TCR sequencing. A substantial number of YFV-reactive T cells expressed memory phenotype markers and contained expanded clones in the absence of exposure to YFV. After vaccination, pre-existing YFV-specific T cell populations with low clonal diversity underwent limited expansion, but rare populations with a reservoir of unexpanded TCRs generated robust responses. These altered dynamics reorganized the immunodominance hierarchy and resulted in an overall increase in higher avidity T cells. Thus, instead of further increasing the representation of dominant clones, YFV vaccination recruits rare and more responsive T cells. Our findings illustrate the impact of vaccines in prioritizing T cell responses and reveal repertoire reorganization as a key component of effective vaccination., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Gout is a common inflammatory arthritis caused by the deposition of monosodium urate (MSU) crystals in the joints. This activates the macrophages into a proinflammatory state by inducing NLRP3-dependent interleukin-1β (IL-1β) secretion, resulting in neutrophil recruitment. Soluble decoy receptor 3 (DcR3) is an immune modulator and can exert biological functions via decoy and non-decoy actions. Previously, we showed that DcR3 suppresses lipopolysaccharides (LPS)- and virus-induced inflammatory responses in the macrophages and promotes the macrophages into the M2 phenotype. In this study, we clarified the actions of DcR3 and its non-decoy action motif h eparin sulfate proteoglycan (HSPG) b inding d omain (HBD) in the MSU crystal-induced NLRP3 inflammasome activation in the macrophages and in mice. In bone marrow-derived macrophages, THP-1 and U937 cells, we found that the MSU crystal-induced secretion of IL-1β and activation of NLRP3 were suppressed by both DcR3.Fc and HBD.Fc. The suppression of the MSU-induced NLRP3 inflammasome activation is accompanied by the inhibition of lysosomal rupture, mitochondrial production of the reactive oxygen species (ROS), expression of cathepsins, and activity of cathepsin B, without affecting the crystal uptake and the expression of NLRP3 or pro-IL-1β. In the air pouch mice model of gout, MSU induced less amounts of IL-1β and chemokines secretion, an increased M2/M1 macrophage ratio, and a reduction of neutrophil recruitment in DcR3-transgenic mice, which expresses DcR3 in myeloid cells. Similarly, the mice intravenously treated with DcR3.Fc or HBD.Fc displayed less inflammation response. These findings indicate that HBD of DcR3 can reduce MSU crystal-induced NLRP3 inflammasome activation via modulation of mitochondrial and lysosomal functions. Therefore, we, for the first time, demonstrate a new therapeutic potential of DcR3 for the treatment of gout., Competing Interests: 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., (Copyright © 2021 Pan, Huang, Sekar, Huang, Lin and Hsieh.)

Introduction: Dystonia is a movement disorder with high clinical and genetic heterogeneity. Mutations in Anoctamin-3 (ANO3) gene have been reported to cause dystonia 24 (DYT24). This study aims to clarify the spectrum and frequency of ANO3 rare variants in Chinese populations with primary dystonia and understand the clinical and genetic features of DYT24., Methods: Sanger sequencing was used to screen all exons and exon-intron boundaries of ANO3 for rare variants in 115 primary dystonia patients. The clinical manifestations of patients with ANO3 variants in our study and previously reported literatures were further characterized., Results: Four distinct variants of ANO3 (c.1127A > G, c.1235 T > A, c.1531-3T > C, c.-11G > T) were identified in six unrelated individuals. Combined with our work and literature review, a total of 35 different rare variants distributed in ANO3 were identified in 62 dystonia patients. The predominant phenotype is cranio-cervical dystonia and more than half of patients develop head/limb tremor. Most of patients presented with isolated dystonia whereas few of them showed combined dystonia. The age of onset ranged from 1 to 69 years and peaked in late adulthood, while for generalized dystonia it peaked in a young age. Half of patients with generalized dystonia experienced deep brain stimulation (DBS). And all of them showed improvement of dystonia by DBS., Conclusions: This study confirms a relatively high frequency of rare ANO3 variants in Chinese patients with dystonia and indicates that the late adulthood-onset, cranio-cervical dystonia seems to be an important feature of the ANO3 phenotype. Further functional studies are warranted to understand the role of ANO3 in dystonia., (Copyright © 2020. Published by Elsevier B.V.)

This work investigated the effects of hot air drying pretreatment (HAD), freeze drying pretreatment (FD) and vacuum drying pretreatment (VD) on the physicochemical properties and structural characterizations of starch isolated from canistels. X-ray diffraction displayed that the starches separated from canistel by different drying pretreatments showed a typical A-type crystal structure. The SEM image showed that cracks and debris appeared on the surface of HVD and VD particles. The molecular structure of starches obtained by different drying pretreatments was studied using Fourier infrared and solid state 13 C CP/MAS NMR analysis. The results indicated that vacuum drying pretreatment could promote the formation of the double helix of starch granules, and hot air drying and freeze drying destroyed the ordered structure of starch granules. These structural changed to affect the physicochemical properties of starch granules. The study of different drying pretreatments to separate starches provided practical value for drying pretreatments. Furthermore, the current study affords information for canistel starches cultivated in China that would be convenient for commercial applications., Competing Interests: Declaration of competing interest The author of this article declares that there is no conflict of interest related to this article., (Copyright © 2020. Published by Elsevier B.V.)

Facial synkinesis can be present in both primary and postparalytic hemifacial spasm (HFS). The present retrospective study aimed to summarize the clinical features of synkinesis and explore an appropriate botulinum toxin A (BoNT-A) injection strategy to manage the synkinesis accompanying HFS. Video recordings of 234 patients with primary and postparalytic HFSs were analyzed. Improvements in the severity of spasm and synkinesis owing to BoNT-A treatment were monitored and compared among 36 primary and 12 postparalytic HFS patients with synkinesis and completed follow-up records. BoNT-A was injected into the voluntary facial region (VFR), the synkinetic facial region (SFR), or both VFR and SFR, and the efficacy of these strategies was evaluated and analyzed. Oral-ocular synkinesis in the primary group (32.8%) and ocular-oral synkinesis in the postparalytic group (81.0%) showed the highest incidence. Patients in both the primary and postparalytic groups exhibited a tremendous alleviation of spasm (97.2% vs. 91.7%, P > 0.05) following BoNT-A treatment. In both groups, coinjection and SFR injection were commonly used and effective in treatment of ocular and oral synkinesis, while VFR was frequently used but ineffective for frontal synkinesis. In addition, the improper muscle selection surrounding the mouth corner resulted in pattern change and treatment failure of oral synkinesis. Synkinesis mostly affected the ocular and oral regions. BoNT-A, via treatment of SFR, is effective against synkinesis accompanying HFS., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: To ascertain if concurrent chemotherapy (CCT) benefits people with stage II nasopharyngeal carcinoma (NPC) treated with two-dimensional radiotherapy (2DRT) or intensity-modulated radiotherapy (IMRT)., Methods: A total of 4157 patients diagnosed with stage II NPC were evaluated. Patients received radiotherapy (RT) with/without CCT. Patients were divided into 2DRT and IMRT subgroups. After propensity score matching, the role of CCT was explored in these two subgroups. Overall survival (OS) was the primary endpoint and progression-free survival (PFS), locoregional relapse-free survival (LRFS) and distant metastasis-free survival (DMFS) were secondary endpoints., Results: In the 2DRT subgroup, CCT addition to RT benefited cases with T1N1/T2N1 in OS, PFS and LRFS (P < .001, P = .003 and P = .003, respectively) significantly, but no difference was observed in patients with T2N0. DMFS were similar in the two arms. CCT was a significant protective factor for OS, PFS, and LRFS for patients with stage N1. In the IMRT subgroup, RT alone could maintain equivalent OS, PFS, LRFS and DMFS (P = .209, .448, .477 and .602 respectively) and cause less acute toxicity compared with concurrent chemoradiotherapy (CCRT)., Conclusion: CCRT was better than 2DRT alone among patients with T1-2N1M0 stage. CCT application for NPC patients receiving IMRT led to no survival benefit and greater toxic effects., (© 2019 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

Hainan/Eksotika papaya is a popular cultivated plant in Hainan Island, China. Papaya seed oil (PSO) contains functional compounds with good antioxidant activity, especially monounsaturated fatty acids. In this work, the ultrasound-assisted extraction (UAE) of PSO was optimized using response surface methodology. It was found that the optimal extraction performance was realized when the elevated time was set to 20 min, the ultrasound power was set to 250 W, and the n-hexane-to-sample ratio was set to 16:1 (v/w). The highest yield of PSO (32.27%) was obtained under the optimal conditions, and PSO showed good oxidative stability. Differential scanning calorimetry analysis showed that the melting point of Hainan/Eksotika PSO was low, while its crystallization temperature was high. FTIR and NMR were used to analyze the chemical structure of PSO, which also proved that PSO possessed good stability without oxidative degradation. In addition, scanning electron micrograph was employed to investigate the change in seed microscopic structure. The results showed UAE caused serious structural damage of sample cell membranes and walls, which help oil access to the solvent with a high extraction ratio. The results indicated that UAE is an efficient environmental-friendly, and promising technique could be applied to produce PSO or other edible oil with a better health-beneficial value in food industry., Competing Interests: There is no conflict of interests regarding the publication of this paper.

Objective: To study the predictive value of procalcitonin as a serum biomarker in diagnosis of late periprosthetic joint infection(PJI) for providing theoretical reference basis for diagnosis of PJI., Methods: A total of 77 cases were retrospective included from January 2015 to December 2017 for revision of total hip arthroplasty and total knee arthroplasty, according to the diagnostic criteria of Musculoskeletal Infection Society(MISI). All cases were divided into infection group and non-infection group. Infection group included 21 cases, 7 cases for male and 14 cases for female, with an average age of (60.70±8.75) years old (ranged 43 to 75 years old). Non-infection group 56 included cases, 24 cases for male and 32 cases for female, with an average age (64.40±12.14) years old (ranged 43 to 85 years old). Concentration of preoperative serum procalcitonin was examined and the chi-square test was used to compare positive rate between the two groups., Results: Two cases in the infection group had positive serum procalcitonin, 0.06 ng/L and 0.10 ng/L respectively, with the positive rate of 9.52%; 4 cases in non-infected group had positive serum procalcitonin, 0.05 ng/L, 0.06 ng/L, 0.06 ng/L, 0.16 ng/L respectively, with the positive rate of 7.14%. No statistically significant difference was observed between the two groups ( P =0.662)., Conclusions: Most of PJI were low toxicity infection with little systemic inflammatory response, so the concentration of serum procalcitonin was normal or slightly higher level, had little clinical significance for diagnosis of PJI. But the cases of this retrospective study are not enough, more cases are needed for further study., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© 2019 by the China Journal of Orthopaedics and Traumatology Press.)

This study evaluated the physicochemical properties of oils extracted from steam-exploded camellia seed ( Camellia oleifera Abel.). Steam pressure, resident time, fatty acid composition, total phenolics, tocopherol, squalene, and sterol contents, and volatile compounds were determined. 1 H NMR and FTIR spectra were performed for the structure of camellia seed oil. This study has found the highest yield of oil was 86.56% and was obtained when steam explosion pretreatment was at 1.6 MPa 30 s. Oil extracted by steam explosion pretreatment exhibited favorable physicochemical properties and stronger antioxidant activities compared to untreated oil. The compositions of fatty acid were similar between treated and untreated camellia seed oil. According to the 1 H NMR and FTIR analyses, the functional groups of the oils were not significantly affected by the steam explosion pretreatment. Furans such as 2-pentyl-furan, 2-furanmethanol, and 3-methyl-furan were produced from stream-exploded camellia seed. Scanning electron microscopy revealed that steam explosion pretreatment efficiently promoted the release of oil by destroying the cell structure of camellia seed. Therefore, steam explosion can be an effective method for the camellia seed oil extraction., Competing Interests: There are no conflicts of interest to declare.

Background: Gold lotion (GL), a natural mixed product made from the peels of six citrus fruits, has recently been identified as possessing anti-oxidative, anti-inflammatory, and immunomodulatory effects. GL has been used to protect skin against UV-induced damage, but its activity against psoriasis, a chronic autoimmune skin disease caused by dysregulation between immune cells and keratinocytes, is not known. We therefore evaluated the effect of GL on imiquimod (IMQ)-induced psoriasis-like inflammation in mice., Results: GL treatment significantly attenuated IMQ-induced psoriasis-like symptoms in mice. The inflammatory cytokines upregulated by IMQ in skin lesions were also inhibited by feeding GL. In addition, GL treatment reduced the infiltration of CD4 + T cells/neutrophils in skin lesions and the percentage of IL-17-/IL-22-producing T cells in lymph nodes. Furthermore, GL impaired IMQ-induced type I interferon production by plasmacytoid dendritic cells (pDCs) in vitro., Conclusion: Our results indicate GL can act to suppress the initiation of psoriasis and strongly suggest that GL may have potential to be applied to the treatment of psoriasis. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

The inhibitory effect of immunoreceptor tyrosine-based activation motif (ITAM)-containing adapters DAP12 and FcεRI γ-chain (FcRγ) has been found in many immune functions. Herein, we have further explored the role of these adapters in C-type lectin receptors response. We identified that FcRγ, but not DAP12, could negatively regulate the Dectin-1 responses in dendritic cells (DCs). Loss of FcRγ or both DAP12 and FcRγ enhanced the maturation and cytokine production in DCs upon Dectin-1 activation compared to normal cells, whereas DCs lacking only DAP12 showed little changes. In addition, increments of T cell activation and T helper 17 polarization induced by FcRγ-deficient DCs were observed both in vitro and in vivo . Examining the Dectin-1 signaling, we revealed that the activations of several signaling molecules were augmented in FcRγ-deficient DCs stimulated with Dectin-1 ligands. Furthermore, we demonstrated that the association of phosphatases SHP-1 and PTEN with FcRγ may contribute to the negative regulation of FcRγ in Dectin-1 activation in DCs. These results extend the inhibitory effect of ITAM-containing adapters to Dectin-1 response in immune functions, even though Dectin-1 contains an ITAM-like intracellular domain. According to the role of Dectin-1 in responding to microbes and tumor cells, our finding may have applications in the development of vaccine and cancer therapy.

Fresh-cut Chinese water chestnuts (CWC) turn yellow after being peeled, reducing their shelf life and commercial value. Metabolomics, the systematic study of the full complement of small molecular metabolites, was useful for clarifying the mechanism of fresh-cut CWC etiolation and developing methods to inhibit yellowing. In this study, metabolic alterations associated with etiolation at different growth stages (0 day, 2 days, 3 days, 4 days, 5 days) from fresh-cut CWC were investigated using LC-MS and analyzed by pattern recognition methods (principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and orthogonal projection to latent structures-discriminant analysis (OPLS-DA)). The metabolic pathways of the etiolation molecules were elucidated. The main metabolic pathway appears to be the conversion of phenylalanine to p -coumaroyl-CoA, followed by conversion to naringenin chalcone, to naringenin, and naringenin then following different pathways. Firstly, it can transform into apigenin and its derivatives; secondly, it can produce eriodictyol and its derivatives; and thirdly it can produce dihydrokaempferol, quercetin, and myricetin. The eriodictyol can be further transformed to luteolin, cyanidin, dihydroquercetin, dihydrotricetin, and others. This is the first reported use of metabolomics to study the metabolic pathways of the etiolation of fresh-cut CWC.

Fresh cut Chinese water-chestnut is a popular ready-to-eat fresh-cut fruit in China. However, it is prone to etiolation and the chemicals responsible for this process are not known yet. To address this problem, we extracted phytochemicals from etiolated Chinese water-chestnut and separated them using MPLC and column chromatography. Four compounds were obtained and their structures were determined by interpretation of UV, TLC, HPLC and NMR spectral data and by comparison with reported data. We identified these compounds as eriodictyol, naringenin, sucrose and ethyl D-glucoside. Among those, eriodictyol and naringenin were both isolated for the first time in fresh-cut Chinese water-chestnut and are responsible for the yellowing of this fruit cutting., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The biological activity of the edible basidiomycete Antrodia cinnamomea (AC) has been studied extensively. Many effects, such as anti-cancer, anti-inflammatory, and antioxidant activities, have been reported from either crude extracts or compounds isolated from AC. However, research addressing the function of AC in enhancing immunity is rare. The aim of the present study is to investigate the active components and the mechanism involved in the immunostimulatory effect of AC. We found that polysaccharides (PS) in the water extract of AC played a major role in dendritic cell (DC) activation, which is a critical leukocyte in initiating immune responses. We further size purified and identified that the high-molecular weight PS fraction (greater than 100 kDa) exhibited the activating effect. The AC high-molecular weight PSs (AC hmwPSs) promoted pro-inflammatory cytokine production by DCs and the maturation of DCs. In addition, DC-induced antigen-specific T cell activation and Th1 differentiation were increased by AC hmwPSs. In studying the molecular mechanism, we confirmed the activation of the MAPK and NF-κB pathways in DCs after AC hmwPSs treatment. Furthermore, we demonstrated that TLR2 and TLR4 are required for the stimulatory activity of AC hmwPSs on DCs. In a mouse tumor model, we demonstrated that AC hmwPSs enhanced the anti-tumor efficacy of the HER-2/neu DNA vaccine by facilitating specific Th1 responses. Thus, we conclude that hmwPSs are the major components of AC that stimulate DCs via the TLR2/TLR4 and NF-κB/MAPK signaling pathways. The AC hmwPSs have potential to be applied as adjuvants.

Hydroxysafflor yellow A (HSYA) is a main bio-active compound important of a traditional Chinese medicine named Carthamus tinctorius L. and has been shown to possess various effects, especially anti-inflammatory benefits and potential protections against acute lung injury (ALI) in previous studies. Therefore, in this present study, we aimed to evaluating effects of HSYA on lipopolysaccharide (LPS)-induced ALI in mice. ALI was induced by intratracheal instillation of LPS into lung, and dexamethasone was used as a positive control. Results demonstrated that HSYA abated LPS-induced pathological change and attenuated lung vascular permeability and edema. HSYA down-regulated both the ability of myeloperoxidase (MPO) in lung tissues and levels of inflammatory mediators including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and IFN(interferon)-β in serum. Moreover, HSYA prevented toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and TIR-domain-containing adapter-inducing interferon-β (TRIF) protein up-expressions. In addition, the activations of mitogen-activated protein kinases including p38, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) were blocked by HSYA. And also, the phosphorylations of interferon regulatory factor 3 (IRF3), translocation of nuclear factor kappa B (NF-κB)/p65 and inhibitory kappa B (IκB)-α were inhibited by HSYA. In conclusion, HSYA attenuated inflammatory response in ALI mice through inhibition of TLR 4-dependent signaling pathways.

Human beings do not passively perceive the facial expressions of other people, but predict observed facial expressions by employing past experiences. The aim of the current study was to investigate whether and how anticipation affected the perception of facial expressions. A 3-way repeated-measures ANOVA on anticipation, orientation, and facial expression was performed on RTs and recognition accuracy in Experiments 1 and 2. The results showed that anticipation reduced susceptibility to negative facial expressions. In this regard, anticipation might be considered as an effective emotion-regulation strategy. In addition, a decreased inversion effect for positive facial expressions was found in the predictable condition, which might reflect a switch from feature-based to holistic processing.

The aim of this study was to determine the value of computed tomography perfusion (CTP) parameters, including cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT) and time-to-peak (TP), in a clinical study of patients with stroke. Additionally, we determined which parameter or combination of parameters are reliable in detecting the presence of an infarct and penumbra. CTP was performed within 24 h of the onset of symptoms in 20 patients with possible stroke. Magnetic resonance imaging (MRI) was performed 3-7 days later and the threshold of the CTP was adjusted according to the results to provide CT images that correlated with the MRI; the MRI results were taken as the gold standard. CBV, CBF and TP contrast agent enhancement were calculated using the CT results. The CTP results were compared with the MRI findings. All CTP parameters were reliable in detecting the penumbra (P<0.001). In these parameters, changes of MTT were the most useful. CTP revealed various changes in CBF, CBV, MTT and TP in ischemic areas. CTP parameters were also reliable in detecting the infarct core (P<0.001). We determined that when detecting the penumbra, all CTP parameters are reliable, and when detecting cerebral ischemia, a combination of parameters should be used.

We report a rare case of a 38-year-old woman with metastatic colonic adenocarcinoma from primary lung adenocarcinoma detected by PET/CT. She underwent colonoscopy and adenocarcinoma was diagnosed in the pathology report, which was the same as that for lymph node biopsy from a left supraclavicular lymph node. We used immunohistochemistry to diagnose primary adenocarcinoma of the lung with colonic metastasis. Owing to mutation in exon 19 of EGFR gene, targeted therapy was given to her with a prescription of oral gefitinib for 1 month as first-line treatment. It was chosen to further treat the patient with chemotherapy and radiotherapy. As the patient was suffering from increasing coughing and sputum, radiotherapy and chemotherapy were subsequently cancelled. Since the general condition of the patient was relatively poor, Tarceva was therefore prescribed. The patient had lived for 5 more months since the diagnosis of metastatic colonic adenocarcinoma.

Background: Cell-based gene therapy using cells that express angiogenic factors is an alternative technique for therapeutic angiogenesis in transplantation of xenogeneic acellular dermis matrix (ADM). However, immune rejection is a substantial obstacle to implantation of genetically engineered allogeneic or xenogeneic cells., Objective: To evaluate application of microencapsulated cells that express vascular endothelial growth factor (VEGF) in xenogeneic ADM transplants to improve wound angiogenesis and healing., Materials and Methods: NIH3T3 cells were genetically modified to secrete VEGF and enveloped in semipermeable microcapsules. Microencapsulated VEGF-NIH3T3 cells were implanted in defects on the dorsa of guinea pigs with xenogeneic ADM and autologous split-thickness skin grafts. Cell structure and microencapsulation were observed at microscopy, and expression of VEGF was detected using an enzyme-linked immunosorbent assay (ELISA) and immunochemistry. Extent of angiogenesis in the ADM and the survival rate of the composite skin were evaluated after 2 weeks. In addition, expression of human VEGF and CD31 in the implanted acellular dermis was assessed, and microvessel density was calculated., Results: Microencapsulated VEGF-expressing NIH3T3 cells were prepared successfully, and demonstrated proliferation and viability, and expressed VEGF both in vitro and in vivo. Extent of angiogenesis and survival rate of the composite skin containing the microencapsulated VEGF-expressing cells were significantly greater than in controls. Microencapsulated VEGF-expressing NIH3T3 cells augmented early angiogenesis in ADM implanted on wound and improved healing., Conclusion: Microencapsulated xenogeneic cell-based gene therapy may be a novel approach to therapeutic angiogenesis in transplantation of xenogeneic ADM skin.

Objective: To observe the influence of recombinant adenovirus-mediated PDE5-shRNAs on the free calcium level in rat penile smooth muscle cells and to explore the feasibility of gene therapy for erectile dysfunction (ED)., Methods: Smooth muscle cells of the rat corpus cavernosum were transfected with constructed rAd-rPDE5-shRNAs and then dyed with the calcium fluorescent probe Fluo-3/AM at 24, 48 and 72 hours. The dynamic changes of the calcium fluorescence intensity were observed under the laser scanning confocal microscope (LSCM). The relative level of intracellular calcium was determined by fluorescence indexes., Results: The fluorescence indexes of calcium at 24, 48 and 72 hours were 829.3 +/- 7.8, 801.5 +/- 9.5 and 856.3 +/- 8.7 in the rAd-rPDES-shRNAs group, significantly lower than in the rAd-mock (1106.3 +/- 10.8, 1121.3 +/- 10.2 and 1058.5 +/- 12.1) and blank control group (1076.6 +/- 9.7, 1133.4 +/- 11.2 and 1104.3 +/- 10.5) (P < 0.05)., Conclusion: Adenovirus mediated shRNAs of the target PDE5 gene can significantly decrease the intracellular calcium level in the smooth muscle cells of the rat corpus cavernosum.

Objective: To investigate the effect of phosphodiesterase type 5 (PDE5) small interfering RNA (siRNA) on cyclic guanosine monophosphatethe (cGMP) in the smooth muscle cells of human corpus cavernosum, and to provide laboratory evidence for the application of the RNA interference (RNAi) technique for the treatment of erectile dysfunction., Methods: The recombinant adenovirus rAd5-shRNA-PDE5A3 expressing three pairs of specific shRNA was constructed successfully. The smooth muscle cells of human corpus cavernosum were divided into an experimental, a negative control and a blank control group, and transfected respectively with rAd5-shRNA-PDE5A3, adenovirus rAd5-mock and phosphate buffered saline. The concentration of cGMP was measured by radioimmunoassay at 24, 48 and 72 hours after transfection, and the effect of rAd5-shRNA-PDE5A3 was detected on the cGMP in the smooth muscle cells of the corpus cavernosum., Results: The cGMP level in the smooth muscle cells of the corpus cavernosum was significantly higher in the rAd5-shRNA-PDE5A3 group than in the rAd5-mock control and blank control groups (P < 0.05), most significantly at 72 hours after transfection., Conclusion: The rAd5-shRNA-PDE5A3 can obviously increase the cGMP level in the smooth muscle cells of human corpus cavernosum, and enhance the inhibition of the PDE5 gene.

Objective: To study the feasibility of inducing the differentiation of rat adipose-derived stem cells (ADSCs) into smooth-muscle-like cells in monolayer culture in vitro., Methods: ADSCs were obtained from the adipose of the inguinal region of the SD rat. A growth curve of the ADSCs was drawn. The fourth passage ADSCs were induced to differentiate into adipocytes with adipogenic inducing fluid and determined by Oil Red O staining, into osteoblasts with osteogenic inducing fluid and determined by Von Kossa staining, as well as into smooth-muscle-like cells with inducing fluid containing beta-mercaptoethanol, and the expression of alpha-smooth muscle actin (alpha-SMA) was detected by the immunohistochemical method., Results: The ADSCs presented spindle and polygon shapes and proliferated rapidly in vitro. The growth curve showed the ADSCs in the logarithmic growth phase two days after subculture. The fourth passage ADSCs exhibited red stained lipid droplets characteristic of adipocytes in the cytoplasm on Oil Red O staining after induction with adipogenic inducing fluid, and calcium nodes on Von Kossa staining after induction with osteogenic inducing fluid. The immunohistochemical results of the ADSCs induced into smooth-muscle-like cells showed that the positive rate of alpha-SMA in the beta-mercaptoethanol induced cells was (29.80 +/- 6.89)%, significantly higher than in the uninduced ones ([2.89 +/- 1.24]%, P < 0.01)., Conclusion: ADSCs displayed obvious characteristics of smooth muscle cells after induction, and could be a new source of cells in the tissue engineering studies of smooth muscle related diseases.

Objective: To investigate the clinical effect of acellular dermal matrix (ADM) combined with auto-skin grafting on deep burn wound ,and the result of long-term follow-up and histological examination., Methods: One hundred and fifty-two patients with deep burn hospitalized from February 2000 to July 2003 were repaired with porcine ADM and auto split-thickness graft. Wound healing rate was assessed 1 week after operation. Degree of cicatricial hyperplasia was examined 1, 3, 6, 12 months after operation. Wound samples from 5 patients were harvested for histological examination 72 months after operation, for which transmission electron microscopy were employed in 2 cases., Results: Grafts completely survived was seen in 116 patients (accounting for 76.3% of cases), survival rate over 95% were observed in 23.7% of cases. One hundred and twenty-seven patients were followed up 1 month after operation, in whom mild local contraction, cord like scar was seen along its junction with skin, its texture was soft ,and there was no pruritus or blister formation. One hundred and one patients were followed up 3 months after operation, and the graft showed mild contraction less marked when compared with that of the site where auto split-thickness skin grafting was used. Articular function was good. Eighty-two patients were followed up 6 months after operation,color and texture of grafts were similar to normal skin with no obvious cicatricial hyperplasia. Fifty-eight patients were followed up 12 months after operation, the texture of grafts was similar to normal skin without obvious reject reaction. Sixteen patients were followed up over 72 months after operation, the grafts appeared dry compared with normal skin. Histological examination showed: tissue structure of grafts was similar to normal skin, intact small sweat gland and sweat gland cells were not found in dermal layer., Conclusion: Heterologous ADM combined with auto split-thickness graft can survive in human body without obvious immune rejection reaction for a long time. No intact small sweat gland or sweat gland cells in dermis is a problem worth of study in regeneration of skin function.

Objective: To investigate the role of xenogenic (porcine) ADM as dermal substitute in scar treatment., Methods: After scar excision, the wounds were covered with composite grafts of DR procine ADM and autologous thin split-thickness grafts in one stage or in two stages., Results: 22 out of 47 cases were treated in two-staged procedure. After the ADMs were applied to the wound, the autologous thin split-thickness grafts were implanted 7 days later. 25 cases were treated in one-staged procedure. The survival rates of composite grafts were (88.3 +/- 3.7)% for subcutaneous recipient bed and (89.7 +/- 3.4)% for deep fascia recipient bed in group with two-staged procedure, compared with (92.5 +/- 4.1)% and (93.2 +/- 5.2)%, respectively, in group with one-staged procedure. Early after grafts taken, the grafts had a pink colour and smooth surface. The patients were followed up for 90 days at most. The survived composite grafts were durable, elastic, smooth and soft with good function and appearance like normal skin. They could even be pinched up. The scar along the edge of the grafts was slightly hypertrophic., Conclusions: The survival rate of composite graft is higher in patients with one-staged procedure. The elasticity and textural of the taken grafts are better on subcutaneous recipient bed than on deep fascia recipient bed, though the function has no difference. Xenogenic (porcine) ADM can be an optimal dermal substitute for wound coverage after scar excision.

Objective: To explore the effect of one dressing of porcine acellular dermal matrix on deep partial thickness burns., Methods: From January 1997 to January 2004, sixty-seven cases of deep partial thickness total burned surface area (TBSA) from 50% to 90% burn wound were treated by a single dressing of porcine acellular dermal matrix (the porcine acellular dermal matrix group). Ten cases of deep partial thickness burned patients with the same TBSA treated by exposure method served as the exposure method group. The healing time of the wound was observed. The patients were followed up for 3 months to 2 years, and the scar proliferation was observed., Results: The deep partial-thickness wound would be healed without dressing change in the porcine acellular dermal matrix group, and the average healing time was (12.2 +/- 2.6) days. The average healing time of the exposure method group was (27.4 +/- 3.5) days. Follow up of the patients within 3 months to 2 years showed that scar proliferation in the porcine acellular dermal matrix group was much less than that in the exposure method group, even no scar proliferation was observed in some patients., Conclusion: Without tangential excision, autografting and dressing change, a single dressing of porcine acellular dermal matrix on deep partial thickness burn wound could shorten the healing time and inhibit scar proliferation.

Objective: To explore a new injectable material., Methods: Micronized swine acellular dermal matrix was injected into the ear of rabbits. The augmentation was observed and measured with pathological and transmission electron microscopic technique., Results: There were fibroblasts and capillaries in the injected material. The absorption rate of injected acellular dermal matrix was 26.93% in the experimental group at 48 weeks after injection., Conclusion: The acellular dermal matrix is an ideal injectable material for lower absorption and easy usage.

Objective: To investigate the possibility of treating deep partial-thickness burns by closed dressing of the wounds with porcine acellular dermal matrix (ADM) and evaluate the therapeutic effects., Methods: We conducted a retrospective review of 128 cases of burn patients who received treatment with porcine ADM within the period from January 1998 to January 2002 in our hospital. Different procedures were adopted according to the degree of the burn injury. As for "fairly superficial" deep partial-thickness skin burns, after removing the necrotic epidermis and washing with 0.1 % benzalkonium bromide, the wound was covered with porcine ADM pretreated with povidone-iodine and then bandaged with cotton gauze and bandages. In cases of "fairly deep" deep partial skin thickness burns, eschar excisions as deep as to expose parabiotic lamina were performed prior to dressing the wounds in the same manner as described above., Results: All the patients were successfully treated with satisfactory clinical results., Conclusions: Porcine ADM is feasible as an efficient dressing material for deep partial-thickness burns, which may promote epithelialization in the wounds and help stabilize the patients' condition during burn shock stage to reduce the complications and shorten the treatment courses.

OBJECTIVE: To observe the prognostic value of intrapartum continuous fetal heart monitoring(FHM). METHODS: A review of 150 patients who had intrapartum FHM was performed. The relation between fetal heart rate and amniotic fluid condition, newborn Apgar scores, and delivery method was determined. RESULTS: Among the 150 cases,normal fetal heart rates were recorded in 108 cases. The abnormalities in the other 42 cases with abnormal FHM were as follows: cloudy amniotic fluid 27 cases(64.3%)compared with 17/108 (15.7%) in the normal FHM group (P<0.005); 10 cases (23.8%) had Apgar scores <8 compared with only 2 (1.85%) in the normal FHM group (P<0.005). In the abnormal FHM group,there were 15 cases (35.7%) of spontaneous labor,10 cases (23.8%) of forceps delivery and 17 cases (40.5%) of C-section compared with 81, 3, and 24 (75%,2.8%, and 22.2%) respectively in the normal FHM group (P<0.005). Spontaneous delivery was more common in the normal FHM (75%)compared with abnormal FHM group(35.7%). C-section rate was higher in the abnormal FHM group (40.5%) (P<0.005). CONCLUSION: Intrapartum fetal heart monitoring detects early fetal distress and thus may lead to earlier intervention such as C-section during the perinatal period.