Halothiobacillus diazotrophicus sp. nov., a chemolithoautotrophic sulphur-oxidizing and nitrogen-fixing bacterium isolated from freshwater.

A sulphur-oxidizing and nitrogen-fixing bacterium, designated strain LS2T, was isolated from freshwater collected from the Pearl River in Guangzhou, PR China. The strain was an obligate chemolithoautotroph, utilizing reduced sulphur compounds (sulphide, sulphite, elemental sulphur, thiosulphate and tetrathionate) as energy sources and electron donors. Diazotrophic growth of strain LS2T was observed at 15-40 °C, pH 5-9, with a NaCl concentration range of 0-0.68 mol l-1 and with oxygen content higher than 21 %. The major cellular fatty acids were summed feature 8 (comprising C18 : 1 ω7c and/or C18 : 1 ω6c) and C16 : 0. The DNA G+C content of the complete genome sequence was 60.7 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain LS2T formed a lineage within the family Halothiobacillaceae, showing gene sequence identity of 96.8 % with its closest relative Halothiobacillus neapolitanus c2. The genome of strain LS2T contains multiple genes encoding sulphur-oxidizing enzymes that catalyse the oxidation of reduced sulphur compounds and an nif complex encoding enzymes for nitrogen fixation. In addition, the genome contains genes encoding cbb3-type cytochrome c oxidase, aa3-type cytochrome c oxidase, bd-type quinol oxidase and cytochrome o oxidase, which enable the survival strain LS2T under oxic and microaerophilic conditions. On the basis of phenotypic, genotypic and phylogenetic data, strain LS2T is considered to represent a novel species of the genus Halothiobacillus, for which the name Halothiobacillus diazotrophicus sp. nov. is proposed. The type strain is LS2T (=GDMCC 1.4095T=JCM 39442T).

A graph-based genome and pan-genome variation of the model plant Setaria.

Setaria italica (foxtail millet), a founder crop of East Asian agriculture, is a model plant for C4 photosynthesis and developing approaches to adaptive breeding across multiple climates. Here we established the Setaria pan-genome by assembling 110 representative genomes from a worldwide collection. The pan-genome is composed of 73,528 gene families, of which 23.8%, 42.9%, 29.4% and 3.9% are core, soft core, dispensable and private genes, respectively; 202,884 nonredundant structural variants were also detected. The characterization of pan-genomic variants suggests their importance during foxtail millet domestication and improvement, as exemplified by the identification of the yield gene SiGW3, where a 366-bp presence/absence promoter variant accompanies gene expression variation. We developed a graph-based genome and performed large-scale genetic studies for 68 traits across 13 environments, identifying potential genes for millet improvement at different geographic sites. These can be used in marker-assisted breeding, genomic selection and genome editing to accelerate crop improvement under different climatic conditions.

A combination of umbilical cord mesenchymal stem cells and monosialotetrahexosy 1 ganglioside alleviates neuroinflammation in traumatic brain injury.

Neuro-inflammation and activated microglia play important roles in neuron damage in the traumatic brain injury (TBI). In this study, we determined the effect of neural network reconstruction after human umbilical cord mesenchymal stem cells (UMSCs) combined with monosialotetrahexosy 1 ganglioside (GM1) transplantation and the effect on the neuro-inflammation and polarization of microglia in a rat model of TBI, which was established in male rats using a fluid percussion brain injury device. Rats survived until day 7 after TBI were randomly treated with normal control (NC), saline (NS), GM1, UMSCs, and GM1 plus UMSCs. Modified neurological severity score (mNSS) was assessed on days 7 and 14, and the brain tissue of the injured region was collected. Immunofluorescence, RT-PCR, and western blot analysis found that inhibitory neuro-inflammatory cytokines TGF-ß and CD163 protein expression levels in injured brain tissues were significantly increased in rats treated with GM1 + UMSCs, GM1, or UMSCs and were up-regulated compared to saline-treated rats. Neuro-inflammatory cytokines IL-6, COX-2 and iNOS protein expressions were down-regulated compared to rats treated with saline. The protein expression levels of NE, NF-200, MAP-2 and ß-tubulin III were increased in the injured brain tissues from rats treated with GM1 + UMSCs, or GM1 and UMSCs alone compared to those in the rats treated with NS. The protein expression levels in rats treated with GM1 plus UMSCs were most significant on day 7 following UMSC transplantation. The rats treated with GM1 plus UMSCs had the lowest mNSS compared with that in the other groups. These data suggest that UMSCs and GM1 promote neural network reconstruction and reduce the neuro-inflammation and neurodegeneration through coordinating injury local immune inflammatory microenvironment to promote the recovery of neurological functions in the TBI.

CASP6 predicts poor prognosis in glioma and correlates with tumor immune microenvironment.

Background: Glioma is an aggressive tumor of the central nervous system. Caspase-6 (CASP6) plays a crucial role in cell pyroptosis and is a central protein involved in many cellular signaling pathways. However, the association between CASP6 and prognosis of glioma patients remains unclear. Methods: Four bioinformatic databases were analyzed to identify differentially expressed genes (DEGs) between glioma and healthy tissues. Eighty-one protein-coding pyroptosis-related genes (PRGs) were obtained from the GeneCards database. The pyroptosis-related DEGs (PRDEGs) were extracted from each dataset, and CASP6 was found to be aberrantly expressed in glioma. We then investigated the biological functions of CASP6 and the relationship between CASP6 expression and the tumor microenvironment and immunocyte infiltration. The half maximal inhibitory concentration of temozolomide and the response to immune checkpoint blockade in the high- and low-CASP6 expression groups were estimated using relevant bioinformatic algorithms. Quantitative real-time reverse transcription PCR and western blotting were carried out to confirm the different expression levels of CASP6 between human astrocytes and glioma cell lines (U251 and T98G). We determined the role of CASP6 in the tumorigenesis of glioma by knocking down CASP6 in U251 and T98G cell lines. Results: We found that CASP6 was overexpressed in glioma samples and in glioma cell lines. CASP6 expression in patients with glioma correlated negatively with overall survival. In addition, CASP6 expression correlated positively with the degree of glioma progression. Functional analysis indicated that CASP6 was primarily involved in the immune response and antigen processing and presentation. Patients with high CASP6 levels responded more favorably to temozolomide, while patients with low expression of CASP6 had a better response to immunotherapy. Finally, in vitro experiments showed that CASP6 knockdown inhibited glioma proliferation. Conclusions: The pyroptosis-related gene CASP6 might represent a sensitive prognostic marker for patients with glioma and might predict their response of immunotherapy and temozolomide therapy. Our results might lead to more precise immunotherapeutic strategies for patients with glioma.

CAMK1D Inhibits Glioma Through the PI3K/AKT/mTOR Signaling Pathway.

Calcium/calmodulin-dependent protein ID (CAMK1D) is widely expressed in many tissues and involved in tumor cell growth. However, its role in gliomas has not yet been elucidated. This study aimed to investigate the roles of CAMK1D in the proliferation, migration, and invasion of glioma. Through online datasets, Western blot, and immunohistochemical analysis, glioma tissue has significantly lower CAMK1D expression levels than normal brain (NB) tissues, and CAMK1D expression was positively correlated with the WHO classification. Kaplan-Meier survival analysis shows that CAMK1D can be used as a potential prognostic indicator to predict the overall survival of glioma patients. In addition, colony formation assay, cell counting Kit-8, and xenograft experiment identified that knockdown of CAMK1D promotes the proliferation of glioma cells. Transwell and wound healing assays identified that knockdown of CAMK1D promoted the invasion and migration of glioma cells. In the above experiments, the results of overexpression of CAMK1D were all contrary to those of knockdown. In terms of mechanism, this study found that CAMK1D regulates the function of glioma cells by the PI3K/AKT/mTOR pathway. In conclusion, these findings suggest that CAMK1D serves as a prognostic predictor and a new target for developing therapeutics to treat glioma.

Association of Single-Nucleotide Polymorphisms of Gab1 Gene with Susceptibility to Meningioma in a Northern Chinese Han Population.

BACKGROUND The Gab1 gene has an important role in cell proliferation in meningioma via various signaling pathways. However, the relationship between polymorphisms of the Gab1 gene and meningioma remains unknown. In this study, we aimed to investigate the plausible association of single-nucleotide polymorphisms (SNPs) of the Gab1 gene and meningioma risk in a northern Chinese Han population. MATERIAL AND METHODS This case-control study included 205 patients with meningioma and 297 healthy controls. Four loci of the Gab1 gene were genotyped using the multiplex snapshot technique. The odds ratio (OR) and 95% confidence interval (CI) were calculated by chi-squared and logistic regression analysis. The distributions of Gab1 SNP genotypes and allele frequencies were compared between patients with meningioma and healthy controls and among patients stratified by clinical phenotypes. RESULTS The allelic frequency distributions of G at rs3805236 and C at rs1397529 were significantly higher in patients with meningioma than in healthy controls. The frequency of the rs3805236-GG and rs1397529-AC genotypes were significantly higher in patients with meningioma than in controls. Furthermore, there was a statistically significant difference between the genotypes of patients versus healthy individuals at rs1397529, according to stratification by dural invasion. The allelic frequency distributions of alleles or genotypes at rs3805246 and rs3828512 were not different in patients with meningioma and healthy controls. CONCLUSIONS The Gab1 gene rs3805236A>G and rs1397529A>C SNPs increased the risk of meningioma in the northern Chinese Han population. Furthermore, rs1397529A>C may be related to enhanced dural invasion in patients with meningioma.

Different Methods of Fabricating Cartilaginous Ear Framework in Children With Microtia According to the Length of the Eighth Costal Cartilage Intraoperatively.

BACKGROUND: There is controversy over the optimal timing of microtia reconstruction. The eighth costal cartilage, which is used to shape the helix framework, can be one of the key factors determining surgical timing of microtia reconstruction. Nevertheless, it is difficult to predict the length of the eighth costal cartilage preoperatively. The aim of the present study was to discuss different methods of fabricating cartilaginous ear framework in children with microtia according to different lengths of the eighth costal cartilage. METHODS: Based on the actual length of the eighth costal cartilage in microtia children, there are 2 methods to fabricate auricular framework. In method I, the eighth costal cartilage was divided into 2 parts. Part A was used to fashion the helix, while part B was used to protrude the antihelix, superior, and inferior crus. The seventh rib was used to form the main body and the sixth rib was used to form the base of the framework. In method II, the seventh costal cartilage was used to fashion the helix and extrude the antihelix, superior, and inferior crus as method I did. The sixth rib was used to form the main body and the eighth rib was used to form the base of the framework. RESULTS: A total of 68 microtia children underwent auricular reconstruction adopting the modified techniques between 2015 and 2016. The great majority of patients (66 patients) were satisfied with the reconstructed ears. Two patients were relatively satisfied with the reconstructed ears. Three cases have been selected to illustrate the favorable result achieved. They revealed that the helix, antihelix, superior crus, and inferior crus all appeared distinct and presented a favorable result of the contour of the reconstructed auricle. CONCLUSIONS: Based on different lengths of eighth costal cartilage in children, different methods of fabricating ear framework make full use of the autogenous costal cartilage and elevate anatomical details, demonstrating that personalized treatment is necessary.

MicroRNA-124 represses wound healing by targeting SERP1 and inhibiting the Wnt/ß-catenin pathway.

BACKGROUND: Wound healing is a complex process which restores cellular structures and tissue layers after their destruction. Accumulating evidence has proven that microRNAs (miRs) are involved in wound healing. OBJECTIVES: The aim of the study was to research the role of miR-124 in wound healing. MATERIAL AND METHODS: Keratinocytes were respectively transfected with miR-124 mimic, scrambled miRNA (a negative control of miR-124 mimic: mimic NC), antisense oligonucleotides against miR-124 (ASOmiR124), or a negative control of ASO-miR-124 (ASO-NC), and then cell viability, colony formation, cell cycle, expression of cell cycle-associated proteins, and collagen content were all evaluated. The target gene of miR-124 was predicted using TargetScan and verified with luciferase assay. Subsequently, the effects of target gene overexpression on cell viability, colony formation and collagen synthesis were all evaluated. Finally, the expression levels of key kinases in the Wnt/ß-catenin pathway were detected using western blot analysis. RESULTS: Cell viability, colony formation, expression levels of cell cycle-associated proteins, and collagen content were all significantly reduced by miR-124 overexpression. As predicted using bioinformatics and validated with luciferase assay, stress-associated endoplasmic reticulum protein 1 (SERP1) is a target gene of miR-124. Meanwhile, the miR-124 mimic-induced decrease in cell viability, colony formation and collagen synthesis was reversed by SERP1 overexpression. Furthermore, the miR-124 mimic obviously upregulated glycogen synthase kinase 3ß (GSK-3ß) while downregulating ß-catenin, T cell transcription factor 4 (TCF-4) and leukemia enhancer factor 1 (LEF-1). Additionally, all the effects of ASO-miR-124 were the opposite of those of the miR-124 mimic. CONCLUSIONS: We found that miR-124 inhibited keratinocyte proliferation, collagen biosynthesis and activation of Wnt/ß-catenin by targeting SERP1.

MicroRNA-17-3p promotes keratinocyte cells growth and metastasis via targeting MYOT and regulating Notch1/NF-κB pathways.

Wound healing is a fundamental biological process to restore skin integrity. The role of microRNAs (miRNAs) during this process remains elusive. Thus, our study aimed to investigate the biological functions and its molecular mechanisms of miR-17-3p in cutaneous wound healing. Human keratinocyte cell line HaCaT was transfected with miR-17-3p mimic, antisense oligonucleotides (ASO)-miR-17-3p and corresponding controls respectively. After transfection, MTT, flow cytometry, qRT-PCR and western blot were performed to analyze cell viability, colony-formation and cell cycle. Next, scratch wound, Transwell and western blot were used to examine cell migration ability. Besides, prediction of miR-17-3p target was performed by TargetScan and microRNA database, dual-luciferase reporter assay, qRT-PCR and western blot. Moreover, the functions of the MYOT on cell proliferation and metastasis were detected by transfection with its expression vector. Signal pathways of Notch1 and NF-κB were performed by qRT-PCR and western blot. Results showed that miR-17-3p overexpression distinctly promoted cell viability, colony formation, arrested cells at G2/M phase, and upregulated cyclin D1, cyclin B1 and CDK2. Simultaneously, miR-17-3p overexpression increased cell migration and downregulated E-cadherin but upregulated vimentin and α-SMA expression. Moreover, MYOT was verified as a target of miR-17-3p, and it remarkably inhibited HaCaT cells proliferation and metastasis. Protein expression levels of cyclin D1, cyclin B1, CDK2, vimentin and α-SMA were downregulated while E-cadherin was upregulated by MYOT. Furthermore, miR-17-3p signally activated Notch1/NF-κB pathways. These data demonstrated that miR-17-3p promoted keratinocyte cells proliferation and metastasis via targeting MYOT and activating Notch1/NF-κB signal pathways in cutaneous wound healing.

Effect of heparin on production of basic fibroblast growth factor and transforming growth factor-beta1 by human normal skin and hyperplastic scar fibroblasts.

Heparin affects both dermal fibroblast proliferation and collagen and may mediate these effects by altering the levels of basic fibroblast growth factor (bFGF) and transforming growth factor-beta1 (TGF-beta1) production as a wound healing modulator. The purpose of this study is to probe the effect of heparin on bFGF and TGF-beta1 production by human normal skin and hyperplastic scar fibroblasts. This research investigates the effect of heparin on bFGF and TGF-beta1 production by human normal skin and hyperplastic scar fibroblasts with exposure to 0, 100, 300, or 600 microg/ml heparin for 24, 48, 72, or 96 hours in a serum-free in vitro model. Levels of bFGF and TGF-beta1 in the supernatants were determined by enzyme-linked immunosorbant assay. All doses of heparin significantly stimulated production of bFGF by normal skin (393% to 1019% increase) and hyperplastic scar fibroblasts (405% to 899% increase) at all time points (P < .05). Heparin (300 and 600 microg/ml) also stimulated TGF-beta1 production by normal skin (26% to 83%) and hyperplastic scar fibroblasts (63% to 85%) with statistical significance (P < .05) at various time points. These effects of heparin on normal skin and hyperplastic scar fibroblasts may have implications for hyperplastic scar formation and wound healing in vivo.

[Study on poly A site gene mutation of transforming growth factor-beta1 receptor type II between multiple and single site keloid].

OBJECTIVE: To investigate the pathogenesy deference between multiple and single site keloid by detecting gene mutation of Poly A site of transforming growth factor-beta1 receptor type II (TbetaR II). METHODS: Collecting 20 keloid samples (6 multiple sites keloid samples and 14 single site keloid samples) and extracting DNA from them; designing and synthesizing the primers of Poly A site, then amplifying T1beta II DNA by PCR, analyzing the single strand conformation polymorphism about the products of PCR. After purifying the product of PCR, the site and type of the mutation rate of Poly A site was sequenced directly on the automatic sequencing equipment. RESULTS: It had been found that the Poly A site of TbetaR II in keloid has deletion mutation, its mutation rate in multiple sites keloid was 50% (3/6), in single site keloid 7.1% (1/14). The mutation rate of Poly A site in multiple sites keloid was significant higher than that in single site keloid (P < 0.05) CONCLUSION: It has been supposed that there are some deference in pathogenesy between the multiple and the single site keloid.