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Biomedical subjects

Jing An

Publications and source records attributed to Jing An.

At least 19 recordsLinked to original sources
PubMed2026

Integrated phenotype, endogenous hormones and transcriptome analysis revealed the mechanism of response of Phoebe bournei seedlings to shade signals.

Understory tree seedlings are subjected to prolonged shading stress imposed by the canopy foliage, which significantly impedes their growth. A hallmark of shaded environments is a reduced red to far-red light ratio (R: FR). This study elucidates the physiological and molecular responses of the endangered tree species Phoebe bournei to shading signals. Seedlings were exposed to white light (control) and simulated shading environments with R: FR ratios of 1.5, 0.8, and 0.2. The findings reveal that an increase in the proportion of far-red light significantly enhances seedling height, root-collar diameter, internode length, petiole length, leaf surface area, and leaf biomass. Differentially expressed genes (DEGs) in each treatment group predominantly enrich pathways associated with hormone signaling, stress responses, and photosynthesis. Validation experiments demonstrate that shading promotes the activity of Rubisco and RCA enzymes, total chlorophyll (Chl) accumulation, and elevated levels of hormones including indole-3-acetic acid (IAA), gibberellic acid (GA3), salicylic acid (SA)/methyl salicylate (MeSA), cytokinins (CK), abscisic acid (ABA), and jasmonic acid (JA). Weighted Gene Co-expression Network Analysis (WGCNA) identifies seven hub genes linked to photosynthesis and plant hormone regulation: MYB, KSC, SUAR, CESA POD, CESA, and SAUR. Collectively, shading signals induce P. bournei seedlings to elongate their stems and petioles, enhance photosynthetic enzyme activity, and accumulate specific hormones, with pertinent genes actively participating in light signal transduction. This research sheds light on the shading response mechanism of P. bournei, providing a robust theoretical framework for the breeding of shade-tolerant trees and the conservation of endangered species.

Transcriptome↗
PubMed2007

Neuronal apoptotic signaling pathways probed and intervened by synthetically and modularly modified (SMM) chemokines.

As the main coreceptors for human immunodeficiency virus type 1 (HIV-1) entry, CXCR4 and CCR5 play important roles in HIV-associated dementia (HAD). HIV-1 glycoprotein gp120 contributes to HAD by causing neuronal damage and death, either directly by triggering apoptotic pathways or indirectly by stimulating glial cells to release neurotoxins. Here, to understand the mechanism of CXCR4 or CCR5 signaling in neuronal apoptosis associated with HAD, we have applied synthetically and modularly modified (SMM)-chemokine analogs derived from natural stromal cell-derived factor-1alpha or viral macrophage inflammatory protein-II as chemical probes of the mechanism(s) whereby these SMM-chemokines prevent or promote neuronal apoptosis. We show that inherently neurotoxic natural ligands of CXCR4, such as stromal cell-derived factor-1alpha or viral macrophage inflammatory protein-II, can be modified to protect neurons from apoptosis induced by CXCR4-preferring gp120(IIIB), and that the inhibition of CCR5 by antagonist SMM-chemokines, unlike neuroprotective CCR5 natural ligands, leads to neurotoxicity by activating a p38 mitogen-activated protein kinase (MAPK)-dependent pathway. Furthermore, we discover distinct signaling pathways activated by different chemokine ligands that are either natural agonists or synthetic antagonists, thus demonstrating a chemical biology strategy of using chemically engineered inhibitors of chemokine receptors to study the signaling mechanism of neuronal apoptosis and survival.

AIDS Dementia Complex↗
PubMed2006

Structure-based virtual screening of chemical libraries for drug discovery.

One of the main goals in drug discovery is to identify new chemical entities that have a high likelihood of binding to the target protein to elicit the desired biological response. To this end, virtual screening is being increasingly used as a complement to high-throughput screening to improve the speed and efficiency of the drug discovery and development process. The availability of inexpensive high-performance computing platforms in recent years has transformed this field into one that is highly diverse and rapidly evolving, where large chemical databases have been successfully screened to identify hits for a wide range of targets such as Bcl-2 family proteins, G protein-coupled receptors, kinases, metalloproteins, nuclear hormone receptors, proteases and many more.

Acetylcholinesterase↗
PubMed2006

A simple method to synthesize triangular silver nanoparticles by light irradiation.

We describe a simple method to synthesize triangular silver nanoparticles by photoreducing the silver ions by citrate. A noteworthy difference of the present method as compared with the previous photo-induced methods is that good shape control over the nanoparticles can be realized in the absence of soft templates or polymer directing agents. The formation process of the silver nanoparticles was investigated by UV-vis spectroscopy and transmission electron microscopy (TEM). It was found that the concentration of reactant plays important role in the morphology control of produced silver nanoparticles. As one of the applications of these nanoparticles, they were used as surface-enhanced Raman scattering substrates and 1,4-bis[2-(4-pyridyl)ethenyl]-benzene (BVPP) was used as a Raman probe to evaluate the enhancement ability of the triangular silver nanoparticles.

Chemistry↗
PubMed2006

SMM-chemokines: a class of unnatural synthetic molecules as chemical probes of chemokine receptor biology and leads for therapeutic development.

Chemokines and their receptors play important roles in numerous physiological and pathological processes. To develop natural chemokines into receptor probes and inhibitors of pathological processes, the lack of chemokine-receptor selectivity must be overcome. Here, we apply chemical synthesis and the concept of modular modifications to generate unnatural synthetically and modularly modified (SMM)-chemokines that have high receptor selectivity and affinity, and reduced toxicity. A proof of the concept was shown by transforming the nonselective viral macrophage inflammatory protein-II into new analogs with enhanced selectivity and potency for CXCR4 or CCR5, two principal coreceptors for human immunodeficiency virus (HIV)-1 entry. These new analogs provided insights into receptor binding and signaling mechanisms and acted as potent HIV-1 inhibitors. These results support the concept of SMM-chemokines for studying and controlling the function of other chemokine receptors.

Amino Acid Sequence↗
PubMed2006

Inhibition of CXCR4 with the novel RCP168 peptide overcomes stroma-mediated chemoresistance in chronic and acute leukemias.

The chemokine receptor CXCR4 mediates the migration of hematopoietic cells to the stroma-derived factor 1alpha (SDF-1alpha)-producing bone marrow microenvironment. Using peptide-based CXCR4 inhibitors derived from the chemokine viral macrophage inflammatory protein II, we tested the hypothesis that the inhibition of CXCR4 increases sensitivity to chemotherapy by interfering with stromal/leukemia cell interactions. First, leukemic cells expressing varying amounts of surface CXCR4 were examined for their chemotactic response to SDF-1alpha or stromal cells, alone or in the presence of different CXCR4 inhibitors. Results showed that the polypeptide RCP168 had the strongest antagonistic effect on the SDF-1alpha- or stromal cell-induced chemotaxis of leukemic cells. Furthermore, RCP168 blocked the binding of anti-CXCR4 monoclonal antibody 12G5 to surface CXCR4 in a concentration-dependent manner and inhibited SDF-1alpha-induced AKT and extracellular signal-regulated kinase phosphorylation. Finally, RCP168 significantly enhanced chemotherapy-induced apoptosis in stroma-cocultured Jurkat, primary chronic lymphocytic leukemia, and in a subset of acute myelogenous leukemia cells harboring Flt3 mutation. Equivalent results were obtained with the small-molecule CXCR4 inhibitor AMD3465. Our data therefore suggest that the SDF-1alpha/CXCR4 interaction contributes to the resistance of leukemia cells to chemotherapy-induced apoptosis. Disruption of these interactions by the peptide CXCR4 inhibitor RCP168 represents a novel strategy for targeting leukemic cells within the bone marrow microenvironment.

Chemokine CXCL12↗
PubMed2006

[Construction of fusion expression vector EGFP-PDX-1 and its transfection into rat fetal hepatic stem cells by electroporation].

OBJECTIVE: To construct the fusion expression vector of pancreatic-duodenal homeobox gene 1 (PDX-1) fused to green fluorescent protein (GFP) capable of stable expression in fetal rat hepatic stem cells after transfection by electroporation. METHODS: PDX-1 cDNA was amplified from SK900/BLSCRIPT plasmid and cloned into the multiple cloning site of pEGFP-C1 to obtain the recombined plasmid pEGFP-C1-PDX-1. Rat fetal hepatic stem cells were isolated, cultured, identified and transfected with the recombinant vector by electroporation, followed by observation of these cells with fluorescent microscope. The result of transfection was analyzed by RT-PCR and cell growth curve. RESULTS: Identification by enzyme digestion confirmed successful construction of the recombinant vector. Fetal hepatic stem cells can stably express GFP and PDX-1 for a period of time, and their growth and proliferation was not obviously affected after transfection. CONCLUSION: The fusion expression vector of EGFP-PDX-1 is successfully constructed and stably expressed in rat fetal hepatic stem cells, which may facilitate the study of the role of PDX-1 in stem cell differentiation into insulin-producing cells.

Animals↗
PubMed2005

Different stereochemical requirements for CXCR4 binding and signaling functions as revealed by an anti-HIV, D-amino acid-containing SMM-chemokine ligand.

Human immunodeficiency virus type 1 (HIV-1) uses a chemokine receptor, usually CXCR4 or CCR5, for entry into the target cells. Here, we used a chemical biology approach to demonstrate that binding and signaling domains in CXCR4 are possibly distinct and separate, as the new analogue, D(1-10)-vMIP-II-(9-68)-SDF-1alpha (RCP222), could not activate CXCR4 despite the fact that its binding activity was comparable to that of stromal cell-derived factor (SDF)-1alpha, the only natural ligand of CXCR4.

Amino Acid Sequence↗
PubMed2005

A simple strategy for generation of gene knockdown constructs with convergent H1 and U6 promoters.

RNA interference (RNAi) is a powerful tool for functional genetic studies in model organisms and mammalian cells. To facilitate rapid construction of gene knockdown constructs and RNAi libraries for known genes of mammalian cells, a new and simple strategy to produce small interfering RNA (siRNA) expression vectors with two opposing polymerase III promoters was developed. The design involved a one-step PCR amplification and single cloning procedure to construct a dual promoter siRNA expression vector. The forward primer is identical for all PCR reactions, only a single reverse primer that contains the siRNA targeting sequence has to be synthesized in the construction of each individual vector. This single primer design is cost-effective and it reduces the risk of sequence errors during synthesis of long oligos. Sense and antisense strands of siRNA duplexes were transcribed from the same template and this eliminated the need to synthesize long hairpin-forming oligonucleotides. Our study demonstrated that this vector design could mediate potent inhibition of expression of both exogenous and endogenous genes in mammalian cells.

Animals↗
PubMed2005

Downregulation of c-myc protein by siRNA-mediated silencing of DNA-PKcs in HeLa cells.

DNA-dependent protein kinase (DNA-PK) has been intensively investigated for its roles in the nonhomologous end-joining (NHEJ) pathway of DNA double-strand break repair and maintenance of genomic stability. Its catalytic subunit, DNA-PKcs, a serine/threonine protein kinase, has recently been reported to be overexpressed in various human cancers, but its significance is unclear. In our study, we synthesized 3 small interfering RNA (siRNA) oligonucleotides, which separately target the translation initiation region, catalytic motif and a sequence between the scid-mutation region and the FATC motif of DNA-PKcs; 3 stable cell lines were generated from HeLa cells transfected with these siRNA constructs, respectively. All 3 siRNAs resulted in remarkable depression on DNA-PKcs expression in HeLa cells, and led to an increased sensitivity to 2 or 4 Gy of gamma-ray as well as 5 or 10 J/m(2) of ultraviolet (UV) irradiation. The siRNA targeting the catalytic motif of DNA-PKcs exhibited the greatest efficiency of radiosensitization. We demonstrated that c-myc protein level was suppressed more than 80% by siRNA-mediated silencing of DNA-PKcs. Using an E-box enhancer (c-myc binding element) driving a secreted alkaline phosphatase (SEAP) reporter strategy, we further found that the transcriptional activity of c-myc was extremely suppressed by silencing DNA-PKcs. The highest suppression effect on c-myc expression was observed in the cells transfected with the siRNA targeting the catalytic motif of DNA-PKcs. Moreover, a similar suppression on c-myc expression and activity was also detected in HeLa cells treated with wortmannin, a phosphatidylinositol (PI)-3 kinase inhibitor. However, silencing DNA-PKcs did not change the level of c-myc mRNA. We have further identified the interaction between DNA-PKcs and c-myc protein. Together, our results imply that DNA-PKcs activity is necessary or contributory to the expression of c-myc protein. Targeting DNA-PKcs is an attractive anticancer strategy, which can achieve through at least two mechanistic pathways: (i) sensitizing cancer cells to radiotherapy or chemotherapy of DNA-damaging agents and (ii) downregulation of c-myc protein.

Androstadienes↗
PubMed2005

Surface-enhanced Raman activity and stability study of silver films prepared by reduction of Ag+ ions in N,N-dimethylformamide.

A new SERS-active silver film prepared by directly depositing silver nanoparticles on glass is reported. The spontaneous reduction of Ag+ ions in N,N-dimethylformamide (DMF) (in the absence of a protecting agent) leads to the deposition of silver nanoparticles on clean glass surfaces in contact with the solution. The observed optical micrographs and AFM images show that the arrangement of the nanoparticles is inhomogeneous and the particles appear to be closely packed. These films are shown to be excellent substrates for SERS measurements, demonstrating significant enhancement, good stability, and trace detection capability. The SERS enhancement of the silver films with different deposition times was compared and found that the 8-h film gives the largest enhancement ability with an enhancement factor estimated at about 10(9). It is also found that the silver films stored for 80 days in air show no significant degradation in their sensitivity.

Dimethylformamide↗
PubMed2005

Magnetic resonance imaging-based spirometry for regional assessment of pulmonary function.

In this work MRI-based spirometry is presented as a method for noninvasively assessing pulmonary mechanical function on a regional basis. A SPAMM tagging sequence was modified to allow continuous dynamic imaging of the lungs during respiration. A motion-tracking algorithm was developed to track material regions from time-resolved grid-tagged images. Experiments were performed to image the lungs during quiet breathing and volumetric strain was calculated from the measured displacement maps. Regional volume calculations, derived from volumetric strain, were integrated over the entire lung and compared to segmented volume calculations with good agreement. Results from this work demonstrate that MRI spirometry has the potential to become a clinically useful tool for measuring regional ventilation and assessing pulmonary diseases that regionally affect the mechanical function of the lung.

Adult↗
PubMed2005

Distinct functional sites for human immunodeficiency virus type 1 and stromal cell-derived factor 1alpha on CXCR4 transmembrane helical domains.

The entry of human immunodeficiency virus type 1 (HIV-1) into the cell is initiated by the interaction of the viral surface envelope protein with two cell surface components of the target cell, CD4 and a chemokine coreceptor, usually CXCR4 or CCR5. The natural ligand of CXCR4 is stromal cell-derived factor 1alpha (SDF-1alpha). Whereas the overlap between HIV-1 and SDF-1alpha functional sites on the extracellular domains of CXCR4 has been well documented, it has yet to be determined whether there are sites in the transmembrane (TM) helices of CXCR4 important for HIV-1 and/or SDF-1alpha functions, and if such sites do exist, whether they are overlapping or distinctive for the separate functions of CXCR4. For this study, by employing alanine-scanning mutagenesis, (125)I-SDF-1alpha competition binding, Ca(2+) mobilization, and cell-cell fusion assays, we found that the mutation of many CXCR4 TM residues, including Tyr(45), His(79), Asp(97), Pro(163), Trp(252), Tyr(255), Asp(262), Glu(288), His(294), and Asn(298), could selectively decrease HIV-1-mediated cell fusion but not the binding activity of SDF-1alpha. Phe(87) and Phe(292), which were involved in SDF-1alpha binding, did not play a significant role in the coreceptor activity of CXCR4, further demonstrating the disconnection between physiological and pathological activities of CXCR4 TM domains. Our data also show that four mutations of the second extracellular loop, D182A, D187A, F189A, and P191A, could reduce HIV-1 entry without impairing either ligand binding or signaling. Taken together, our first detailed characterization of the different functional roles of CXCR4 TM domains may suggest a mechanistic basis for the discovery of new selective anti-HIV agents.

Binding Sites↗
PubMed2005

Unique ligand binding sites on CXCR4 probed by a chemical biology approach: implications for the design of selective human immunodeficiency virus type 1 inhibitors.

The chemokine receptor CXCR4 plays an important role as the receptor for the normal physiological function of stromal cell-derived factor 1alpha (SDF-1alpha) and the coreceptor for the entry of human immunodeficiency virus type 1 (HIV-1) into the cell. In a recent work (S. Tian et al., J. Virol. 79:12667-12673, 2005), we found that many residues throughout CXCR4 transmembrane (TM) and extracellular loop 2 domains are specifically involved in interaction with HIV-1 gp120, as most of these sites did not play a role in either SDF-1alpha binding or signaling. These results provided direct experimental evidence for the distinct functional sites on CXCR4 for HIV-1 and the normal ligand SDF-1alpha. To further understand the CXCR4-ligand interaction and to develop new CXCR4 inhibitors to block HIV-1 entry, we have recently generated a new family of unnatural chemokines, termed synthetically and modularly modified (SMM) chemokines, derived from the native sequence of SDF-1alpha or viral macrophage inflammatory protein II (vMIP-II). These SMM chemokines contain various de novo-designed sequence replacements and substitutions by d-amino acids and display more enhanced CXCR4 selectivity, binding affinities, and/or anti-HIV activities than natural chemokines. Using these novel CXCR4-targeting SMM chemokines as receptor probes, we conducted ligand binding site mapping experiments on a panel of site-directed mutants of CXCR4. Here, we provide the first experimental evidence demonstrating that SMM chemokines interact with many residues on CXCR4 TM and extracellular domains that are important for HIV-1 entry, but not SDF-1alpha binding or signaling. The preferential overlapping in the CXCR4 binding residues of SMM chemokines with HIV-1 over SDF-1alpha illustrates a mechanism for the potent HIV-1 inhibition by these SMM chemokines. The discovery of distinct functional sites or conformational states influenced by these receptor sites mediating different functions of the natural ligand versus the viral or synthetic ligands has important implications for drug discovery, since the sites shared by SMM chemokines and HIV-1 but not by SDF-1alpha can be targeted for the development of selective HIV-1 inhibitors devoid of interference with normal SDF-1alpha function.

Anti-HIV Agents↗
PubMed2005

[Effects of modification of the nutritional components in potassium simplex optimized medium on preimplantation development and cleavage of mouse embryos in vitro].

OBJECTIVE: To solve the problem known as 2-cell block in in vitro culture of preimplantation embryos, modification of the nutritional components in potassium simplex optimized medium (KSOM) was attempted and the preimplantation development and cleavage of mouse embryos in such medium observed. METHOD: One-cell mouse embryos collected from the oviduct of the superovulated mice were cultured in microdrops of different culture media for 5 days [144 h after human chorionic gonadotrophin (hCG) injection], and the percentages of embryos developed to pre-blastocyst, unhatched blastocyst, partially hatched blastocyst and completely hatched blastocyst were recorded at deferent time points (96, 120 and 144 h post-hCG) and total cell number of blastocysts at 120 h post-hCG counted. With these indices as the evaluation criteria, we evaluated the modifications of the concentrations of glucose (0.2, 5.56 mmol/l), bovine serum albumin (BSA, 1 and 4 mg/ml), and amino acids (1/2X NEAA and EAA, or not) in KSOM for their effect on the development of Kunming mouse zygotes in vitro. RESULTS: Increasing the concentration of glucose and BSA was not shown to have significant effects on the rate of blastocyst formation at 120 h post-hCG or the total cell counts in KSOM without amino acids. After supplementation of the medium with amino acids, the rate of partially and completely hatched blastocyts was significantly increased in KSOM or KSOM supplemented with high concentrations of glucose and BSA (KSOMGB). Glucose at the high concentration of 5.56 mmol/L did not inhibit the development of mouse zygotes to hatched blastocysts cultured in KSOM supplemented with amino acids. CONCLUSION: Increasing the concentration of glucose and BSA and amino acids in KSOM can promote the preimplantation development of mouse zygotes in in vitro culture.

Amino Acids↗
PubMed2005

[Experimental study of in situ electroporation-induced testis injury in specific pathogen-free male Kunming mice].

OBJECTIVE: To observe testis injuries induced by in situ electroporation in specific pathogen-free (SPF) adult male Kunming mice. METHODS: With two sets of parameters of high and low voltages, respectively, in situ electroporation of the testis was performed in vivo by fixing the testis and epididymis of the mice between a pair of rectangular tweezer-type electrodes. Two weeks after electroporation, the mice were killed and the testis and epididymis separated and fixed with 4% paraformaldehyde after recording the weight of the testis. Routine histological sections were prepared and observed under optical microscope after HE staining. The epididymis was transferred into M16 medium for spermatozoa separation, and after dilution of the spermatozoa suspension, the spermatozoa viability was observed under optical microscope. RESULTS: Two weeks after high-voltage electroporation, examination of spermatozoa viability and microscopy of the testis sections revealed irreversible testis injury, and the testis weight was significantly reduced in comparison with that of control mice (P<0.01). Low-voltage electroporation, in contrast, only caused reversible injuries of the testis, and the male mice retained their reproductive capacity after a certain length of recovery period. The testis weight after low-voltage electroporation showed no significant difference from that of the control mice (P>0.05). CONCLUSIONS: Appropriate setting of the parameters for in vivo electroporation may avoid severe impact on the reproductive capacity of the testis in SPF male Kunming mice. This technique also provides a possibility for exogenous gene transfer into the reproductive cells.

Animals↗
PubMed2005

Silencing of DNA-PKcs alters the transcriptional profile of certain signal transduction genes related to proliferation and differentiation in HeLa cells.

DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a member of a sub-family of phosphoinositol 3-kinases, has been reported overexpressed in various human cancers, but its significance is unclear. In the present study, we generated the stable cell line HeLa(siRNAH1) of silenced DNA-PKcs by transfecting HeLa cells with the siRNA construct targeting the catalytic motif of DNA-PKcs. The expression of DNA-PKcs was markedly suppressed in HeLa(siRNAH1) cells, and eventuating in increased cellular sensitivity to ionizing radiation as well as cisplatin. Microarray assay was used to explore the transcriptional profiling of signal transduction-associated genes. The results demonstrated that 15 genes were up-regulated and eight were down-regulated in HeLa(siRNAH1) as compared with the HeLa(control) cells that transfected with non-specific siRNA construct. Seven of the up-regulated genes are associated with the interferon-signaling events, the others function in the BMP signal pathway, or as regulators of cell cycle and differentiation. The down-regulated genes include IL8, IL10RA, DAPK3, and those involved in nuclear factor of activated T cells (NFAT) signal pathway and endocrine responsiveness. Using the NFAT-driving secreted alkaline phosphatase reporter expression system, we further confirmed that NFAT transcriptional activity was markedly minimized after silencing DNA-PKcs. These results demonstrated that inactivation of DNA-PKcs altered the transcriptional level of certain signal transduction-associated genes related to proliferation and differentiation.

Cell Differentiation↗