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Biomedical subjects

Chao Gao

Publications and source records attributed to Chao Gao.

14 recordsLinked to original sources

Integrated single-cell and bulk transcriptomic analysis identifies a novel senescent fibroblast subtype associated with poor prognosis in acral melanoma.

BACKGROUND: Acral melanoma (AM) exhibits significant intratumoral heterogeneity, but its tumor microenvironment (TME) and immune regulation remain unclear. This study aims to dissect TME heterogeneity and establish a prognostic model based on key cell subpopulations. METHODS: We collected AM single-cell RNA sequencing (scRNA-seq) and bulk RNA-seq data from the Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA). Unsupervised clustering, CellChat, and Scissor analysis were performed to characterize cellular heterogeneity, cell-cell communication, and prognosis-related cell subpopulations. Kaplan-Meier analysis was used to assess the prognostic value of key genes, which were further validated by multiplex immunohistochemistry (mIHC). RESULTS: In AM, Mel_C2, C7, and C9 with high SEMA6A and KIT expression were strongly linked to poor prognosis. We further identified a senescent fibroblast subpopulation (sCAF_CDKN2A) characterized by high fibroblast senescence signature (FSS) scores. Integrating Scissor analysis of fibroblast subtypes with bulk prognostic data, we identified COL3A1, VCAN, and KIT as prognosis-associated genes upregulated in poor-outcome-related fibroblast subsets. Cell-cell communication analysis revealed that sCAF_CDKN2A engages in an immunosuppressive network, interacting with regulatory T cells (Tregs) via MIF signaling and receiving signals from exhausted CD8+ T cells through PPIA-BSG interactions. Using transcription factor expression patterns from these fibroblast subtypes, we constructed a prognostic model that effectively stratified patients into distinct risk groups with significant differences in overall survival (OS). mIHC confirmed significantly higher protein levels of SEMA6A and COL3A1 in tumor tissues compared to matched normal tissues. CONCLUSIONS: We established a novel prognostic model for AM and identified sCAF_CDKN2A as an immunosuppressive senescent fibroblast subpopulation driving poor prognosis.

Acral melanoma↗

Pseudomonas stutzeri as a novel biocatalyst for pyruvate production from DL-lactate.

Pseudomonas stutzeri SDM oxidized DL-lactic acid (25.5 g l(-1)) into pyruvic acid (22.6 g l(-1)) over 24 h. Both NAD(+)-independent D-lactate dehydrogenase and NAD(+)-independent L-lactate dehydrogenase were found for the first time in the bioconversion of lactate to pyruvate based on the enzyme activity assay and proteomic analysis.

Biotransformation↗

Magnetic carbon nanotubes: synthesis by electrostatic self-assembly approach and application in biomanipulations.

Magnetic multiwalled carbon nanotubes (MWNTs) were facilely prepared by the electrostatic self-assembly approach. Poly(2-diethylaminoethyl methacrylate) (PDEAEMA) was covalently grafted onto the surfaces of MWNTs by MWNT-initiated in situ atom transfer radical polymerization (ATRP) of 2-diethylaminoethyl methacrylate (DEAEMA). The PDEAEMA-grafted MWNTs were quaternized with methyl iodide (CH(3)I), resulting in cationic polyelectrolyte-grafted MWNTs (MWNT-PAmI). Magnetic iron oxide (Fe(3)O(4)) nanoparticles were loaded onto the MWNT surfaces by electrostatic self-assembling between MWNT-PAmI and Fe(3)O(4), affording magnetic nanotubes. The assembled capability of the nanoparticles can be adjusted to some extent by changing the feed ratio of Fe(3)O(4) to MWNT-PAmI. The obtained magnetic nanotubes were characterized with TEM, EDS, STEM, and element mapping analyses. TEM and EDS measurements confirmed the nanostructures and the components of the resulting nanoobjects. The magnetic nanotubes were assembled onto sheep red blood cells in a phosphate buffer solution, forming magnetic cells. The blood cells attached with or without magnetic nanotubes can be selectively manipulated in a magnetic field. These results promise a general and efficient strategy to magnetic nanotubes and the fascinating potential of such magnetic nanoobjects in applications of bionanoscience and technology.

Animals↗

[The influence of Ad-AVEGF165 on human malignant melanoma growth in nude mice].

OBJECTIVE: To investigate the effect of antisense VEGF165 infection on the growth of A375 cells in nude mice. METHODS: A375 cells were injected s.c into the axilla of the nude mouse. After the tumor formed, we cut it into 16 pieces equally, then transplanted into another 15 nude mice. There were three groups: Group PBS, Group Ad-GFP, and Group Ad-aVEGF. Four weeks after interfere, the mice were sacrificed and their tumors were excised for naked eye and histological observation. The VEGF expression was checked with ISH and immunohistochemistry staining. The micro-vessel density (MVD) in tumor mass was counted by VIII factor immunohistochemistry staining. RESULTS: The visible and palpable nodules had developed at all the injected sites. Tumor growth speed was more slowly in Group Ad-aVEGF than that in other groups. GFP gene could express effectively in tumor mass. Ad-aVEGF infection could suppress the growth of tumors, and there were no obvious side effects. Ad-aVEGF resulted more tissue necrosis, but it had no obvious effect on cell apoptosis. VEGF expression was inhibited significantly in Group Ad-aVEGF, and MVD was decreased accordingly. CONCLUSIONS: Ad-aVEGF interfere may be a new method against human malignant melanoma, whose main mechanism is to induce ischemia, but not apoptosis.

Adenoviridae↗

[Temporal and spatial distribution characteristics of nitrogen losses in hilly area of Taihu Lake].

Four typical land uses in hilly area of Taihu Lake were studied on temporal and spatial distribution characteristics of nitrogen losses in surface runoff under natural rainfall through experiment in situ. The medium value of event mean concentration (EMC) of ammonia nitrogen (NH4(+) -N) which dominated agricultural N in surface runoff accounted for 44.5% of total nitrogen (TN), while nitrite nitrogen (NO2(-) -N) accounted for 1.8%. Concentration of nitrogen in runoff had significantly seasonal variation which was related to meteorologic conditions such as rainfall, temperature, and agricultural activities. Temporal variabilities of site mean concentration (SMC) for TN, NH4(+) -N, NO3(-) -N and NO2(-) -N were decreased sequentially. The highest SMC value of TN, NH(+) -N and NO3(-) -N in upland runoff and N2(-) -N in bamboo grove were observed. The spatial distribution of nitrogen losses was determined by fertilizer application and vegetation coverage. Spatial variabilities for SMC of nitrogen were less than temporal variabilities. It was found that transportation fluxes of nitrogen in surface runoff from upland and vegetable plot were higher than that from chestnut orchard and bamboo forest which have significantly related to surface runoff volume.

China↗

Polyurea-functionalized multiwalled carbon nanotubes: synthesis, morphology, and Raman spectroscopy.

An in situ polycondensation approach was applied to functionalize multiwalled carbon nanotubes (MWNTs), resulting in various linear or hyperbranched polycondensed polymers [e.g., polyureas, polyurethanes, and poly(urea-urethane)-bonded carbon nanotubes]. The quantity of the grafted polymer can be easily controlled by the feed ratio of monomers. As a typical example, the polyurea-functionalized MWNTs were measured and characterized in detail. The oxidized MWNTs (MWNT-COOH) were converted into acyl chloride-functionalized MWNTs (MWNT-COCl) by reaction with neat thionyl chloride (SOCl2). MWNT-COCl was reacted with excess 1,6-diaminohexane, affording amino-functionalized MWNTs (MWNT-NH2). In the presence of MWNT-NH2, the polyurea was covalently coated onto the surfaces of the nanotube by in situ polycondensation of diisocyanate [e.g., 4,4'-methylenebis(phenylisocyanate)] and 1,6-diaminohexane, followed by the removal of free polymer via repeated filtering and solvent washing. The coated polyurea content can be controlled to some extent by adjusting the feed ratio of the isocyanato and amino groups. The structure and morphology of the resulting nanocomposites were characterized by FTIR, NMR, Raman, confocal Raman, TEM, EDS, and SEM measurements. The polyurea-coated MWNTs showed interesting self-assembled flat- or flowerlike morphologies in the solid state. The signals corresponding to that of the D and G bands of the carbon nanotubes were strongly attenuated after polyurea was chemically tethered to the MWNT surfaces. Comparative experiments showed that the grafted polymer species and structures have a strong effect on the Raman signals of polymer-functionalized MWNTs.

Journal Article↗

[Effect of a novel splicing mutation (IVS2-2A-->C) of SEDL gene on RNA processing].

X-linked spondyloepiphyseal dysplasia tarda (SEDL) is a rare osteochondrodysplasia caused by the mutation of SEDL gene, which mainly involves vertebral bodies and hips. To explore the effect of the novel splicing mutation (IVS2 -2A-->C) of SEDL gene on mRNA processing in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to elucidate the molecular base of SEDL, total RNA was isolated from EDTA blood samples of patients and controls. RT-PCR was performed on total RNA. cDNA was analyzed by bi-directionally direct sequencing of PCR products and Polyacrylamide gel electrophoresis (PAGE). Sequencing analysis revealed that there were two kinds of cDNA in patients. One is that exon 2 directly spliced exon 4, that is, exon 3 absence from the mature mRNA; and the other is that exon 1 directly spliced exon 4, meaning both exon 2 and 3 being spliced out completely. Meanwhile one kind of cDNA that exon 1 directly spliced exon 3 was found in normal controls. By PAGE, RT-PCR amplified products, 679bp and 537bp, were detected in normal controls, while 567bp and 425bp fragments were found in affected individuals. Our data show that the mutation of the splice-acceptor site in intron 2 causes exon 3 entirely exclusion from the mature RNA transcripts in affected individuals. As the translation start site of the SEDL gene locates on exon 3, the splicing defect causes affected individuals failure to produce sedlin, which elucidates the causative role of SEDL gene in the pathogenesis of SEDL. The absence of exon 2 indicates that there is alternative splicing in SEDL gene. The alternative splicing was also found in normal controls, which demonstrated that the alternative splicing might not be related to the phenotype of SEDL. Because the alternative splicing of exon 2 occurred in the 5'UTR, it is not clear whether it affects the gene expression.

Base Sequence↗

Controlled functionalization of multiwalled carbon nanotubes by in situ atom transfer radical polymerization.

The in situ ATRP (atom transfer radical polymerization) "grafting from" approach was successfully applied to graft poly(methyl methacrylate) (PMMA) onto the convex surfaces of multiwalled carbon nanotubes (MWNT). The thickness of the coated polymer layers can be conveniently controlled by the feed ratio of MMA to preliminarily functionalized MWNT (MWNT-Br). The resulting MWNT-based polymer brushes were characterized and confirmed with FTIR, 1H NMR, SEM, TEM, and TGA. Moreover, the approach has been extended to the copolymerization system, affording novel hybrid core-shell nanoobjects with MWNT as the core and amphiphilic poly(methyl methacrylate)-block-poly(hydroxyethyl methacrylate) (PMMA-b-PHEMA) as the shell. The approach presented here may open an avenue for exploring and preparing novel carbon nanotubes-based nanomaterials and molecular devices with tailor-made structure, architecture, and properties.

Journal Article↗

[Relationship of plasma nitric oxide concentration and the activity of nitric oxide synthase with coronary heart disease].

OBJECTIVE: To study the levels of nitric oxide synthase (NOS) and nitric oxide (NO) in patients with coronary heart disease (CHD), and the factors affecting them. METHODS: Concentrations of NO and the activities of NOS in plasma in 136 patients and 206 controls using the corresponding Kits were measured. The data were analyzed using covariance and multiple linear regression analysis with SAS 8.1. RESULTS: The levels of NO [(217.05 +/- 153.31) micromol/L] and NOS [(14.09 +/- 7.14) U/ml] in patients were significantly higher than that in the healthy controls [(140.69 +/- 90.96) micromol/L, (7.75 +/- 3.79) U/ml, respectively, P < 0.01]. Smoking and drinking were the independent risk factors for NOS, while sex was the independent risk factor for NO (P < 0.01). CONCLUSIONS: The levels of plasma NO and NOS are closely related with coronary heart disease.

Adult↗

[Identification of a novel mutation IVS2-2A-->C of SEDL gene in a Chinese family with X-linked spondyloepiphyseal dysplasia tarda].

OBJECTIVE: To identify the mutation of spondyloepiphyseal dysplasia tarda (SEDL) gene in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to make a discussion on the pathogenesis of SEDL at the molecular level. METHODS: In two patients, four exons comprising the SEDL open reading frame as well as their exon/intron boundaries were analyzed by bi-directional direct sequencing of PCR products. The sequencing results were compared against the normal sequences in GenBank to find the mutation. Then the mutation was identified in other members of the family. RESULTS: A nucleotide substitution of the splice acceptor in SEDL intron 2, IVS2 -2A-->C,was detected in two affected individuals (IV(15) V(3)) in the Chinese family with SEDL, but no sequence change occurring on exons 3-6 was detected. The transversion was also identified in four heterozygous carriers. The mutation was not found in two unaffected male individuals and fifteen normal controls. Furthermore, four potential carriers were identified in the family. CONCLUSION: The mutation IVS2 -2A-->C of SEDL gene was firstly determined in the world. The change of the splice acceptor in SEDL intron 2 may cause skipping of exon 3 which is responsible for the disease. Molecular diagnosis can be made by detecting the mutation.

Alternative Splicing↗

[Gene diagnosis of X-linked spondyloepiphyseal dysplasia tarda by linkage analysis and DNA sequencing].

OBJECTIVE: X linked spondyloepiphyseal dysplasia tarda (SEDL) is heritable osteochondrondysplasia characterized in affected males by disproportional short stature with short neck and trunk resulting from a growth defect of the vertebral bodies, accompanied by barrel chest and degenerative osteoarthropathy of hip joints. This progressive skeletal dysplasia is caused by the SEDL gene located approximately 100 kb centromeric of DXS16 at Xp22. The disorder usually manifests in late childhood without systemic complications, and generally female carriers of SEDL are asymptomatic. So the diagnosis of potential carriers and presymptomatic patients is almost impossible. This study aimed to establish methods of gene diagnosis for finding out potential carriers and presymptomatic patients. METHODS: The blood samples were collected from 21 individuals in a large Chinese pedigree with SEDL. Microsatellite marker DXS16 was selected for linkage analysis. In order to confirm the allele of DXS16 linked to the pathogenic SEDL gene, polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE) were used to examine the variability of the lengths of DXS16, and linkage analysis was performed for the diagnosis of potential carriers and presymptomatic patients. Then the pathogenic mutation of the SEDL gene in the family was identified by bi-directionally direct sequencing of PCR products amplified for each of the four coding exons as well as their exon/intron boundaries. The potential carriers and presymptomatic patients were also diagnosed in this way. RESULTS: Six young individuals (IV(14), IV(19), IV(21), IV(23), V(4), V(7))who wanted to know whether they were carriers or presymptomatic patients were diagnosed by linkage analysis. Four females of them (IV(14), IV(19), IV(21), V(7)) were determined being carriers because they carry the allele of DXS16 which links the pathogenic SEDL gene, and the other two (IV(23), V(4)) being normal individuals for their alleles of DXS16 linked with wild SEDL gene. DNA sequencing identified that the pathogenic mutation of SEDL gene in the family, which was a nucleotide substitution of the splice-acceptor site in intron 2, IVS2 -2 A-->C. This is a novel mutation in the SEDL gene. Four female individuals (IV(14), IV(19), IV(21), V(7)) carried the mutation; individuals IV(23) and V(4) carried the wild SEDL gene. The results of diagnosis of linkage analysis coincide completely with that of DNA sequencing. CONCLUSION: Linkage analysis is a simple, rapid and inexpensive gene diagnosis method for SEDL and its accuracy was the same as DNA sequencing. Each of linkage analysis and DNA sequencing can be used to diagnose SEDL, which is very helpful for finding potential carriers and presymptomatic patients.

Base Sequence↗

Effects of shenmai injection on expression of TNF-alpha mRNA in peritoneal macrophages of scald mice.

OBJECTIVE: To explore the effect of shenmai injection (SI) on expression of TNF-alpha mRNA in peritoneal macrophages (pMPhis) of scald mice. METHODS: BALB/c mice were inflicted with 11% of body surface area III degree scald and injected intraperitoneally (ip) with SI daily for 5 days, and expression of TNF-alpha mRNA in pMPhis was determined by semi-quantitative RT-PCR. RESULTS: In scald mice, the expression of TNF-alpha mRNA in pMPhis increased significantly, but it was reduced obviously (P < 0.01) after SI administration, while the livability was increased markedly (P < 0.05). CONCLUSIONS: For scald mice, the cause of death at early stage might be related to the high expression of TNF-alpha mRNA in pMPhis and the use of SI can decrease the death rate.

Animals↗

[Cloning cDNA encoding for GAGA-like element binding proteins in human Jurkat cells].

OBJECTIVE: To explore GAGA-like element binding protein in human cells. METHODS: Yeast one-hybrid system was used to screen the GAGA-like element binding proteins in HTLV-1 transformed Jurkat cell cDNA fusion library. Total RNA extracted from Jurkat cells was first labeled by reverse transcription, and was taken as cDNA probe to hybridize with the candidate positive clones. RESULTS: 9 positive clones were obtained, and 6 out of the 9 clones were positively hybridized with the cDNA probe. CONCLUSIONS: 6 candidate clones encoding for GAGA-like element binding proteins were obtained from Jurkat cells for further investigation.

Carrier Proteins↗

Water-soluble degradable hyperbranched polyesters: novel candidates for drug delivery?

A novel approach to hyperbranched polymers is presented in this work. Hyperbranched polyesters with a large amount of terminal hydroxyl groups are prepared by a one-pot synthesis from commercially available AB-type and CD(n)-type monomers (n >/= 2). In this paper, Michael addition of diethanolamine (CD(2)) or N-methyl-d-glucamine (CD(5)) to methyl acrylate (AB) generates dominantly AD(n)-type intermediates. Further self-condensation of intermediates at higher temperature and in the presence of catalyst gives hyperbranched polyesters. Because of the tertiary amino groups in the backbone and the hydroxyl groups in the linear and terminal units, the resulting hyperbranched polyester is highly soluble in water. Furthermore, the hyperbranched polymer is degradable because of its ester units. So, the water-soluble hyperbranched polyesters might be applied as a novel material for drug delivery.

Biodegradation, Environmental↗