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Biomedical subjects

Hong Lin

Publications and source records attributed to Hong Lin.

At least 19 recordsLinked to original sources

The association between GLP-1R expression and cardiovascular-kidney-metabolic-related diseases in non-diabetic and non-obese population: evidence triangulation using Mendelian randomization, observational and polygenic score association analysis.

BACKGROUND: Glucagon-like peptide-1 receptor (GLP-1R) agonists are emerging as promising therapies for cardiovascular-kidney-metabolic (CKM) related diseases in individuals with type 2 diabetes mellitus (T2DM) or obesity. But their effects in non-obese and non-diabetic individuals are unclear. This study triangulates evidence using Mendelian randomization (MR), polygenic scores (PGS) and observational analyses to estimate the associations of GLP-1R expression with chronic kidney disease (CKD), heart failure (HF) and metabolic dysfunction-associated steatotic liver disease (MASLD). METHODS: For the MR analysis, instruments mimicking GLP-1R expression were identified using pancreas-specific cis-expression quantitative trait loci from GTEx (N ≤ 305). MR-Robust method was used as the primary MR approach. PGS and observational analyses were performed both in non-diabetic and non-obese individuals separately. A genome-wide association study (GWAS) for MASLD (14,231 cases and 348,091 controls) was performed in the general population using data from UK Biobank. RESULTS: GLP-1R expression showed robust effects on CKD (odds ratio [OR] 0.96, 95%CI 0.95 to 0.97, q = 1.7 × 10- 10 ), HF (OR = 0.96, 95%CI 0.94 to 0.97, q = 2.5 × 10- 8) and MASLD (OR = 0.96, 95%CI 0.93 to 0.98, q = 1.3 × 10- 3) in the general population. Consistent results were observed in validation analyses. Furthermore, PGS and observational analyses among non-T2DM and non-obese individuals found little evidence to support its association with CKD, HF or MASLD. GWAS analysis identified eight conditionally independent variants associated with MASLD, in which rs563199662 was a new signal located at TFPI region. CONCLUSIONS: This study provides multilayered evidence for GLP-1R expression in mitigating CKD, HF and MASLD risks in the general population, while de-prioritized its effect on CKM-related diseases in non-obese and non-diabetic individuals. Further clinical trials are needed to validate the effects of GLP-1R agonists in relative health population.

Humans↗

Enhancement of umbilical cord blood cell hematopoiesis by maitake beta-glucan is mediated by granulocyte colony-stimulating factor production.

Maitake beta-glucan (MBG) is an extract from the fruit body of the Grifola frondosa mushroom that is being widely used to treat cancer in Asia. We have previously reported that MBG enhances mouse bone marrow cell (BMC) hematopoiesis in vitro and protects BMC from doxorubicin (DOX) toxicity. In the current study, we investigated the ability of MBG to enhance hematopoiesis and to reduce the toxic effects of DOX on fresh human umbilical cord blood (CB) cells. MBG treatment significantly enhanced the colony formation unit (CFU) response of granulocytes-macrophages (CFU-GM response) over the whole dose range of 12.5 to 100 microg/ml (P < 0.05). The addition of MBG to DOX-treated CB cells significantly protected granulocyte-macrophage colony formation from the toxicity of DOX, which otherwise produced strong hematopoietic repression. MBG also partially replaced recombinant human granulocyte colony-stimulating factor (rhG-CSF), as shown by a significant augmentation of the CFU-GM response in the absence of rhG-CSF. We found that MBG induces granulocyte colony-stimulating factor (G-CSF) production in CB CD33+ monocytes, as detected by intracellular cytokine flow cytometric assessment. In contrast, we found that adult peripheral blood monocytes did not produce a significant G-CSF response to MBG, whereas both adult and CB monocytes produced G-CSF in response to lipopolysaccharide. These studies provide the first evidence that MBG induces hematopoietic stem cell proliferation and differentiation of CFU-GM in umbilical CB cells and acts directly to induce G-CSF.

Fetal Blood↗

A change in conformational dynamics underlies the activation of Eph receptor tyrosine kinases.

Eph receptor tyrosine kinases (RTKs) mediate numerous developmental processes. Their activity is regulated by auto-phosphorylation on two tyrosines within the juxtamembrane segment (JMS) immediately N-terminal to the kinase domain (KD). Here, we probe the molecular details of Eph kinase activation through mutational analysis, X-ray crystallography and NMR spectroscopy on auto-inhibited and active EphB2 and EphA4 fragments. We show that a Tyr750Ala gain-of-function mutation in the KD and JMS phosphorylation independently induce disorder of the JMS and its dissociation from the KD. Our X-ray analyses demonstrate that this occurs without major conformational changes to the KD and with only partial ordering of the KD activation segment. However, conformational exchange for helix alphaC in the N-terminal KD lobe and for the activation segment, coupled with increased inter-lobe dynamics, is observed upon kinase activation in our NMR analyses. Overall, our results suggest that a change in inter-lobe dynamics and the sampling of catalytically competent conformations for helix alphaC and the activation segment rather than a transition to a static active conformation underlies Eph RTK activation.

Amino Acid Substitution↗

Analysis of the genome-wide variations among multiple strains of the plant pathogenic bacterium Xylella fastidiosa.

BACKGROUND: The Gram-negative, xylem-limited phytopathogenic bacterium Xylella fastidiosa is responsible for causing economically important diseases in grapevine, citrus and many other plant species. Despite its economic impact, relatively little is known about the genomic variations among strains isolated from different hosts and their influence on the population genetics of this pathogen. With the availability of genome sequence information for four strains, it is now possible to perform genome-wide analyses to identify and categorize such DNA variations and to understand their influence on strain functional divergence. RESULTS: There are 1,579 genes and 194 non-coding homologous sequences present in the genomes of all four strains, representing a 76. 2% conservation of the sequenced genome. About 60% of the X. fastidiosa unique sequences exist as tandem gene clusters of 6 or more genes. Multiple alignments identified 12,754 SNPs and 14,449 INDELs in the 1528 common genes and 20,779 SNPs and 10,075 INDELs in the 194 non-coding sequences. The average SNP frequency was 1.08 x 10(-2) per base pair of DNA and the average INDEL frequency was 2.06 x 10(-2) per base pair of DNA. On an average, 60.33% of the SNPs were synonymous type while 39.67% were non-synonymous type. The mutation frequency, primarily in the form of external INDELs was the main type of sequence variation. The relative similarity between the strains was discussed according to the INDEL and SNP differences. The number of genes unique to each strain were 60 (9a5c), 54 (Dixon), 83 (Ann1) and 9 (Temecula-1). A sub-set of the strain specific genes showed significant differences in terms of their codon usage and GC composition from the native genes suggesting their xenologous origin. Tandem repeat analysis of the genomic sequences of the four strains identified associations of repeat sequences with hypothetical and phage related functions. CONCLUSION: INDELs and strain specific genes have been identified as the main source of variations among strains, with individual strains showing different rates of genome evolution. Based on these genome comparisons, it appears that the Pierce's disease strain Temecula-1 genome represents the ancestral genome of the X. fastidiosa. Results of this analysis are publicly available in the form of a web database.

Analysis of Variance↗

The endo-beta-mannanase gene families in Arabidopsis, rice, and poplar.

Mannans are widespread hemicellulosic polysaccharides in plant cell walls. Hydrolysis of the internal beta-1,4-D: -mannopyranosyl linkage in the backbone of mannans is catalyzed by endo-beta-mannanase. Plant endo-beta-mannanase has been well studied for its function in seed germination. Its involvement in other plant biological processes, however, remains poorly characterized or elusive. The completed genome sequences of Arabidopsis (Arabidopsis thaliana), rice (Oryza sativa), and poplar (Populus trichocarpa) provide an opportunity to conduct comparative genomic analysis of endo-beta-mannanase genes in these three species. In silico sequence analysis led to the identification of eight, nine and 11 endo-beta-mannanase genes in the genomes of Arabidopsis, rice, and poplar, respectively. Sequence comparisons revealed the conserved amino acids and motifs that are critical for the active site of endo-beta-mannanases. Intron/exon structure analysis in conjunction with phylogenetic analysis implied that both intron gain and intron loss has played roles in the evolution of endo-beta-mannanase genes. The phylogenetic analysis that included the endo-beta-mannanases from plants and other organisms implied that plant endo-beta-mannanases have an ancient evolutionary origin. Comprehensive expression analysis of all Arabidopsis and rice endo-beta-mannanase genes showed divergent expression patterns of individual genes, suggesting that the enzymes encoded by these genes, while carrying out the same biochemical reaction, are involved in diverse biological processes.

Amino Acid Sequence↗

MMP-2 mediates ethanol-induced invasion of mammary epithelial cells over-expressing ErbB2.

Ethanol is a tumor promoter and may enhance the metastasis of breast cancer. We have previously demonstrated that over-expression of ErbB2 promoted ethanol-mediated invasion of mammary epithelial cells and breast cancer cells. However, the underlying cellular/molecular mechanisms remain unknown. By gelatin zymography, we showed that over-expression of ErbB2 increased the production of matrix metalloproteinase-2 (MMP-2) and MMP-9 in human mammary epithelial cells (HB2). Transient or stable transfection of ErbB2 cDNA to HB2 cells upregulated the transcripts and the activity of the MMP-2/-9 gene promoter; the upregulation of MMP-2/-9 expression was mediated by p38 mitogen-activated protein kinase (p38 MAPK) and phosphatidylinositol 3-kinase (PI3K). Although ethanol, at physiologically relevant concentrations (100-400 mg/dl), did not affect the production of MMP-2/-9, it activated MMP-2 in HB2 cells over-expressing ErbB2 (HB2(ErbB2)), but not HB2 cells; it enhanced the cleavage of proform MMP-2 (72 kDa) to an active form (62 kDa). The activation was dependent on c-jun N-terminal kinases (JNKs) and reactive oxygen species (ROS). On the other hand, ethanol affected neither the expression nor the activation of MMP-9. Selective inhibitors of MMP-2 (SB-3CT and OA-Hy) and antioxidants significantly inhibited ethanol-stimulated invasion of HB2(ErbB2) cells. Furthermore, knocking down MMP-2 by small interference RNA also induced a partial blockage on ethanol-promoted invasion of HB2(ErbB2) cells. Thus, ethanol-stimulated invasion of cells over-expressing ErbB2 was mediated, at least partially, by MMP-2 activation.

Blotting, Western↗

Production of Alexa Fluor 488-labeled reovirus and characterization of target cell binding, competence, and immunogenicity of labeled virions.

Respiratory enteric orphan virus (reovirus) has been used to study many aspects of the biology and genetics of viruses, viral infection, pathogenesis, and the immune response to virus infection. This report describes the functional activity of virus labeled with Alexa Fluor 488, a stable fluorescent dye. Matrix assisted laser desorption-time of flight analysis indicated that Alexa Fluor 488 labeled the outer capsid proteins of reovirus. Labeled virus bound to murine L929 fibroblasts as determined by flow cytometry and fluorescence microscopy, and the specificity of binding were demonstrated by competitive inhibition with non-labeled virus. Labeled reovirus induced apoptosis and cytopathic effect in infected L929 cells. Mice infected with labeled virus mounted robust serum antibody and CD8(+) T-cell responses, indicating that labeled virus retained immunogenicity in vivo. These results indicate that Alexa Fluor 488-labeled virus provides a powerful new tool to analyze reovirus infection in vitro and in vivo.

Animals↗

An expanded WW domain recognition motif revealed by the interaction between Smad7 and the E3 ubiquitin ligase Smurf2.

Smurf2 is an E3 ubiquitin ligase that drives degradation of the transforming growth factor-beta receptors and other targets. Recognition of the receptors by Smurf2 is accomplished through an intermediary protein, Smad7. Here we have demonstrated that the WW3 domain of Smurf2 can directly bind to the Smad7 polyproline-tyrosine (PY) motif. Of particular interest, the highly conserved WW domain binding site Trp, which interacts with target PY motifs, is a Phe in the Smurf2 WW3 domain. To examine this interaction, the solution structure of the complex between the Smad7 PY motif region (ELESPPPPYSRYPMD) and the Smurf2 WW3 domain was determined. The structure reveals that, in addition to binding the PY motif, the WW3 domain binds six residues C-terminal to the PY motif (PY-tail). Although the Phe in the WW3 domain binding site decreases affinity relative to the canonical Trp, this is balanced by additional interactions between the PY-tail and the beta1-strand and beta1-beta2 loop of the WW3 domain. The interaction between the Smurf2 WW3 domain and the Smad7 PY motif is the first example of PY motif recognition by a WW domain with a Phe substituted for the binding site Trp. This unusual interaction allows the Smurf2 WW3 domain to recognize a subset of PY motif-containing proteins utilizing an expanded surface to provide specificity.

Amino Acid Motifs↗

Role of neurofilament aggregation in motor neuron disease.

A major question in the pathogenesis of motor neuron disease is why motor neurons are selectively susceptible to mutations in widely expressed gene products. Reexamination of motor neuron degeneration due to alterations of neurofilament (NF) expression suggests that disruption of assembly with aggregation of the light neurofilament (NFL) protein may be an upstream event and contributing factor leading to the preferential degeneration of motor neurons. The implications of these findings are that aggregation of NFL is not only a triggering mechanism to account for the hallmark aggregates of NF protein in sporadic and familial forms of amyotrophic lateral sclerosis, but that aggregates of NFL may also promote aggregation of wildly expressed proteins that are destabilized by missense mutations, such as by mutations in superoxide dismutase-1 protein. This review examines the potential role of NFs in determining and promoting the preferential degeneration of motor neurons in motor neuron disease. The underlying premise is that motor neurons are selectively susceptible to alterations in NF expression, that alterations in NF expression lead to NF aggregates in motor neurons, and that elevated levels of NF aggregates provide a favorable microenvironment for the formation of neurotoxic aggregation and degeneration of motor neurons.

Animals↗

Differentiation and quantitative determination of surface and hydrate water in lyophilized mannitol using NIR spectroscopy.

Mannitol hydrate is a metastable form produced during lyophilization. It is unstable, and therefore can undergo dehydration to release water to the surrounding environment at room temperature. The analysis of this form is challenging due to its thermodynamic instability. This study describes the development of a fast and non-invasive method to determine the mannitol hydrate and surface water content in a lyophilized product using near-infrared (NIR) spectroscopy. The mannitol hydrate was produced through lyophilization and characterized using XRPD, TGA, and NIR spectroscopy. Quantitative methods for hydrate and surface water were developed for NIR spectra with curve fitting and partial least square (PLS) regression models. The curve fitting method deconvoluted the NIR spectra into hydrate and surface water peaks and generated a calibration model by correlating pure spectra peak area to concentration. The standard error of prediction (SEP) for hydrate and surface water content were 0.65 and 0.40%, respectively. The PLS model developed for the same sample set was better than the curve fitting model; SEP = 0.50% for hydrate water and 0.22% for surface water, respectively. The methods can be used to monitor the formation and stability of mannitol hydrate in mannitol-containing formulations during the lyophilization process.

Data Interpretation, Statistical↗

Effect of age and heart rate on atrial mechanical function assessed by Doppler tissue imaging in healthy individuals.

BACKGROUND: The potential usefulness of Doppler tissue imaging (DTI) to assess atrial mechanical function in cardiac disease has been demonstrated. However, there are few reports on normal values of atrial function by DTI analysis. METHODS: Echocardiography with color-coded DTI was performed in 131 healthy control subjects. The peak atrial contraction velocity (V(A)) and the timing of mechanical events were assessed offline at the left atrial (LA) and right atrial (RA) free wall and interatrial septum. RESULTS: V(A) was higher in the RA (9.0 +/- 2.6 cm/s) than the LA (7.5 +/- 2.4 cm/s, P < .001), and both sites were higher than the interatrial septum (5.6 +/- 1.3 cm/s, both P < .001). The interatrial delay was 24 +/- 21 milliseconds. V(A) at the LA was higher in the participants aged 60 years or older than those who were younger than 60 years (8.1 +/- 2.7 vs 6.7 +/- 1.4 cm/s, P < .001), as was the velocity at the RA (9.6 +/- 2.8 vs 8.0 +/- 2.1 cm/s, P < .01). V(A) at the LA was higher in the participants with heart rate of 60/min or higher than those with lower heart rate (7.9 +/- 2.5 vs 6.8 +/- 1.8 cm/s, P < .05). Sex difference had no effect on V(A) and timings of atrial events. CONCLUSIONS: Assessment of atrial mechanical function by DTI is feasible in healthy individuals. The V(A) is the highest at the RA, followed by LA, and the lowest at the interatrial septum. Older age and faster heart rate seems to augment V(A) in the atrial walls.

Aging↗

Expression and functional analysis of the rice plasma-membrane intrinsic protein gene family.

Plasma membrane intrinsic proteins (PIPs) are a subfamily of aquaporins that enable fast and controlled translocation of water across the membrane. In this study, we systematically identified and cloned ten PIP genes from rice. Based on the similarity of the amino acid sequences they encoded, these rice PIP genes were classified into two groups and designated as OsPIP1-1 to OsPIP1-3 and OsPIP2-1 to OsPIP2-7 following the nomenclature of PIP genes in maize. Quantitative RT-PCR analysis identified three root-specific and one leaf-specific OsPIP genes. Furthermore, the expression profile of each OsPIP gene in response to salt, drought and ABA treatment was examined in detail. Analysis on transgenic plants over-expressing of either OsPIP1 (OsPIP1-1) or OsPIP2 (OsPIP2-2) in wild-type Arabidopsis, showed enhanced tolerance to salt (100 mM of NaCl) and drought (200 mM of mannitol), but not to salt treatment of higher concentration (150 mM of NaCl). Taken together, these data suggest a distinct role of each OsPIP gene in response to different stresses, and should add a new layer to the understanding of the physiological function of rice PIP genes.

Abscisic Acid↗

Heroin-administered mice involved in oxidative stress and exogenous antioxidant-alleviated withdrawal syndrome.

Heroin has been shown to elevate dopamine (DA) level. It is well known that an increase in DA oxidative metabolism leads to increased reactive oxygen species (ROS) formation, and thus, ROS have been frequently associated with neuronal cell death due to damage to carbohydrates, amino acids, phospholipids, and nucleic acids. This study investigated whether there are oxidative stress and effects of exogenous antioxidants in heroin-administered mice. The heroin-dependent mice model was made via intraperitoneal injection. Oxidative damage of DNA, protein, and lipid was measured by analysis of single cell electrophoresis, the 2,4-dinitrophenylhydrazine method, and thiobarbituric acid method respectively. The activities of antioxidative enzymes and total antioxidant capacity were assayed by spectrophotometry. After administration with heroin, the mice not only showed decrease of total antioxidant capacity in serum and antioxidant enzymes such as superoxide dismutase, catalase, and glutathione (GSH) peroxidase in brain, but also exhibited the oxidative damages of DNA, protein and lipid. On the other hand, exogenous antioxidants could restrain the oxidative stress, even alleviate withdrawal syndrome in heroin-administered mice. Our results also imply a possibility that ROS may participate in the whole process of dependence and withdrawal of heroin. Therefore, strategies of blocking oxidative stress may be useful in the development of therapy for opiate abuse.

Animals↗

Effect of high intensity ultrasound on the allergenicity of shrimp.

The tropomyosin fraction of shrimp proteins is potentially responsible for allergic reaction in individuals with genetic predisposition to allergy. However, there are no efficient and safe methods to reduce its allergenicity. High intensity ultrasound is known to change the structure of proteins. This study is aimed at assessing high intensity ultrasound's effect on the allergenicity of shrimp allergen. Shrimp and purified shrimp allergen were treated with high intensity ultrasound for 30-180 min. Extracts of treated samples were analyzed by enzyme-linked immunosorbent assay (ELISA) with pool serum of shrimp allergy patients and polyclonal anti-allergen antibodies and by immunoblotting after polyacrylamide gel electrophoresis. Shrimp treated with high intensity ultrasound showed a decrease in allergenicity measured with ELISA. A linear relationship between the immune response induced by treated shrimp allergen and the applied treatment time was observed. The decrease in allergenicity was confirmed by immunoblot assays with shrimp allergic patients serum. Allergenicity of shrimp allergen extracted from treated shrimp was higher than that of purified shrimp allergen with the same treatment time. Gel-filtration HPLC was applied for analysis of shrimp allergen after treatment with high intensity ultrasound. Some fractions were appeared with increasing treatment time. The results suggested that high intensity ultrasound could be used to reduce the allergenicity of shrimp.

Adolescent↗

[Study on cell senescence induced by D-galactose in cultured rat mesenchymal stem cells].

OBJECTIVE: To study morphological and biological senescence changes induced by D-galactose in the cultured rat mesenchymal stem cells. METHODS: After 3rd generations cultured in the DMEM-F12, MSCs were changed into DMEM-F12 medium containing 8 g/L D-galactose and cultured to the 6th generations as the inducement group. The comparison were the 6th generations which was cultured in the DMEM-F12 medium all along, and then identified by surface wave. Using flow cytometer to check the comparisons cell cycle change after swing in with 8 g/L D-galactose within the 4 days. In the first 7 days to draw the growth curve to the two groups. Optical and electronic microscope were used to identify the influences of characteristic morphological of mesenchymal stem cells of the two groups, the influences of biological markers were identified by single cell gel electrophoresis and beta-galactose dye. Results After treatment with D-galactose, the mesenchymal stem cells displayed morphological and biological changes in the cell senescence with the senescent characteristic morphological markers; 85% of the cells were X-gal dye masculine, and the signal cell gel electrophoresis showed DNA damnification. The flow cytometry showed that 90% of the cells stayed in G0/G1, but the cells in S and G2/M almost disappeared. However, the cells in the control group had no such DNA damages. CONCLUSION: D-galactose can induce senescence of the mesenchymal stem cells, and 8 g/L is the best concentration to do so. This study has provided a good model for the research of the mesenchymal stem cells senescence.

Animals↗

[Study on solid phase extraction and spectrophotometric determination of cadmium with 2-[2-(5-bromoquinolinylazo)]-5-diethylaminophenol].

A new chromogenic reagent 2-[2-(5-bromoquinolinylazo)]-5-diethylaminophenol (5-Br-QADEAP) was synthesized, and its structure was verified by elemental analysis, infrared spectrum, and 1H nuclear magnetic resonance spectrum. The color reaction of 5-Br-QADEAP with cadmium was studied. In the presence of pH 8.0 buffer solution and Triton X-100 medium, 5-Br-QADEAP can react with cadmium(II) to form a stable 2 : 1 complex (5-Br-QADEAP: Cd(II). The molar absorptivity is 1.60 x 10(5) L x mol(-1) x cm(-1) at 595 nm. Beer's law is obeyed in the range of 0-1.0 microg x mL(-1). The cadmium ion in the samples can be enriched and separated by solid phase extraction with anion exchange resin cartridge. This method was applied to the determination of cadmium in water and food. The relative standard deviations are 2.5%-3.2%, and the recoveries are 96%-105%. The results are satisfactory.

Animals↗

Determination of trace copper by solid substrate-room temperature phosphorescence quenching method based on activating effect of alpha,alpha'-dipyridyl on Vitamin C reducing beryllon.

A new solid substrate-room temperature phosphorescence quenching method for the determination of trace copper has been established. It is based on the fact that beryllon (R) can emit strong and stable solid substrate-room temperature phosphorescence on the filter paper, and Vitamin C (Vc) reduces R to non-phosphorescent compound that leads to solid substrate-room temperature phosphorescence (SS-RTP) quenching of R, and alpha,alpha'-dipyridyl can activate copper catalyzing Vitamin C reducing R. The DeltaI(p) of the system with alpha,alpha'-dipyridyl is 3.3 times higher than that without alpha,alpha'-dipyridyl, which shows the reaction of alpha,alpha'-dipyridyl activating copper catalyzing Vitamin C reducing R. The reducing value of phosphorescence intensity (DeltaI(p)) is directly proportional to the content of Cu(II) in the range of 0.040-4.0 fg spot(-1) (corresponding concentration: 0.10-10.0 pg ml(-1), sample volume: 0.40 microlspot(-1)). The regression equation of working curve can be expressed as DeltaI(p)=69.99+41.00 m Cu(2+) (fg spot(-1)) (r=0.9980, n=6), and the detection limit is 0.0088 fg spot(-1)(corresponding concentration: 2.2 x 10(-14) g ml(-1)). This sensitive and accurate method with good repeatability and high selectivity has been applied to the determination of trace copper in real samples with satisfactory results. The reaction mechanism for the determination of trace copper by solid substrate-room temperature phosphorescence quenching method based on the activating effect of alpha,alpha'-dipyridyl on Vitamin C reducing beryllon is also discussed.

Ascorbic Acid↗