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Biomedical subjects

Ming Ma

Publications and source records attributed to Ming Ma.

At least 19 recordsLinked to original sources

Proteomics-based analysis of the defense mechanisms of disease-resistant grass carp against Aeromonas veronii.

Sustainable aquaculture of grass carp (Ctenopharyngodon idella, GC) is consistently threatened by bacterial diseases, particularly those caused by Aeromonas veronii. A disease-resistant grass carp (DR-GC) has been developed by backcrossing female gynogenetic GC with normal male GC, exhibiting improved resistance. However, the systemic molecular mechanisms of DR-GC defending against Aeromonas veronii infection remain largely unexplored. Here, a label-free quantitative proteomics approach was employed to systematically compare proteomic profiles across five tissues (intestine, liver, muscle, skin, and kidney) in DR-GC and GC under healthy and infected conditions. The intestine was identified as the central defense tissue, exhibiting the highest number of differentially abundant proteins (DAPs). In DR-GC, A0A3N0YEK7 (small ribosomal subunit protein eS28), A0A3N0YGT8 (ATP synthase-coupling factor 6) and A0A3N0YNS7 (apolipoprotein A-I) were significantly upregulated in intestine, while D5KZW6 (GCHV-induced protein), A0A3N0Z0A1 and Q8JH84 (hemoglobin subunit alpha) were significantly dysregulated across multiple tissues, which playing the critical roles in defense mechanisms at the protein level. Furthermore, cytochrome P450-associated pathways, cytosolic DNA-sensing and RIG-I-like receptor signaling pathways were identified as crucial coordinators mediating immune and metabolic responses. This study provides the first comprehensive proteomic view of multi-tissue defense mechanisms in DR-GC, and identifies key DAPs and pathways for subsequent functional validation.

Animals↗

Development and evaluation of an efficient HPLC/MS/MS method for the simultaneous determination of pseudoephedrine and cetirizine in human plasma: application to phase-I pharmacokinetic study.

A sensitive, simple and highly selective liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and evaluated to determine simultaneously the concentrations of pseudoephedrine and cetirizine in human plasma. The chief benefit of the present method is the minimal sample preparation, as the procedure is only one-step protein precipitation. Two drugs were separated on a C(8) column and analyzed by LC/MS/MS using positive electrospray ionisation (ESI). The method had a chromatographic run time of 12.0 min and a linear calibration curve over the concentration range of 1.0-800 ng/ml for pseudoephedrine and 1.0-400 ng/ml for cetirizine, respectively. The lower limit of quantification of the two drugs was 1.0 ng/ml, respectively. The intra- and inter-batch precisions were less than 9.7%. The method described herein has been first used to reveal the pharmacokinetic characters in healthy Chinese volunteers treated with oral administration of different dosages of cetirizine dihydrochloride and controlled-released pseudoephedrine hydrochloride compound tablet, and approached the influence of a standard meal on the extent and rate of absorption of the combination tablet.

Adult↗

Real-time imaging of nitric oxide production in living cells with 1,3,5,7-tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacence by invert fluorescence microscope.

Although the importance of nitric oxide (NO) as a signalling molecule in many biological processes is becoming increasingly evident, many proposed and potential biological functions of NO still remain unclear. Bioimaging is a good technique to visualize observation of nitric oxide in biological samples. In this report, a fluorescent probe, 1,3,5,7-tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacence (TMDCDABODIPY), has been first applied to real-time image NO produced in PC12 cells, Sf9 cells and human vascular endothelial cells at the presence of l-arginine with inverted fluorescence microscope. NO production in the cells is successfully captured and imaged with fine temporal and spatial resolution. The results prove that the probe combined with inverted fluorescence microscope can be developed into a sensitive and selective method for further study of NO release from cells.

Animals↗

Direct detection of nitric oxide in human blood serum by use of 1,3,5,7-tetramethyl-8-(3',4'-diaminophenyl)difluoroboradiaza-s-indacene with HPLC.

To study the relationship between amounts of nitric oxide (NO) in blood and the development of ischemic cardio-cerebrovascular diseases, trace NO in human blood serum has, for the first time, been determined by use of a 1,3,5,7-tetramethyl-8-(3',4'-diaminophenyl)difluoroboradiaza-s-indacene (TMDABODIPY)-based HPLC method. The proposed method is simple, rapid, and efficient, owing to its high sensitivity and good selectivity for NO. TMDABODIPY and its NO derivative are separated to baseline in 4 min, with simple separation conditions, on a C(18) column eluted with 50 mmol L(-1) ethanolamine in methanol. The derivative is detected by fluorescence at an emission wavelength of 507 nm with excitation at 498 nm. The response is a linear function of concentration in the range 0.8-800 nmol L(-1) NO. The detection limit can reach 2 x 10(-11) mol L(-1) (signal-to-noise ratio=3). The method has been used to detect NO in the serum of patients with five kinds of ischemic cardio-cerebrovascular disease and two diseases closely connected with ischemic cardio-cerebrovascular diseases. Recoveries of NO from spiked serum samples were between 96.58 and 105.71% and concentrations of NO observed in real samples were at 10(-7) mol L(-1) levels. Our studies indicate that the proposed TMDABODIPY-based HPLC technique can be developed into a sensitive and new method for clinical assay and pathology research.

Aza Compounds↗

In vitro labeling and MRI of mesenchymal stem cells from human umbilical cord blood.

OBJECTIVE: The aim of this study was to label human umbilical cord blood mesenchymal stem cells (MSCs) with poly-l-lysine (PLL)-conjugated superparamagnetic iron oxide particles and to obtain magnetic resonance (MR) images of the labeled MSCs' suspension at 1.5 T. MATERIAL AND METHODS: PLL was conjugated with iron oxide to form superparamagnetic particles called Fe(2)O(3)-PLL. Human umbilical cord blood MSCs were isolated, purified, expanded and incubated with Fe(2)O(3)-PLL. Prussian blue stain was performed to show intracellular iron; spectrometry was used to quantify iron uptake within cells. Tetrazolium salt (MTT) assay was applied to evaluate toxicity and proliferation of MSCs labeled with various concentrations of Fe(2)O(3)-PLL. The cell apoptosis rate was determined by annexin V/propichium iodide (PI) double staining method. Vials containing cells underwent MR imaging (MRI) with T(1), T(2) and T(2)* weighted MRI. RESULTS: Iron-containing intracytoplasmatic vesicles could be observed clearly with Prussian blue staining in all samples except the unlabeled control. The iron content per cell determined by spectrometry was 64.51+/-10.32 pg. Among MSCs with and without labeling of various concentrations of Fe(2)O(3)-PLL, MTT values of light absorption had no statistically significant difference (Kruskal-Wallis test, chi(2)=10.35, P=.17). A concentration at 20 mug/ml of iron appeared most suitable for incubating cells. Of labeled and unlabeled MSCs, the early [annexin V-fluorescein isothiocyanate (FITC)-positive/PI-negative] and late (annexin V-FITC-positive/PI-positive) apoptotic cells were 10.34+/-0.43%/11.36+/-1.30% and 4.01+/-1.76%/2.98+/-1.37%, respectively, and there were no significant differences between them (P>.05). T(2) weighted image (WI) and T(2)*WI demonstrated significant decrease of signal intensity (SI) in vials containing 1 x 10(6) (1 day), 1x10(6) (8 days) and 5 x 10(5) labeled cells, in comparison with unlabeled cells (P<.05). The percentage change of SI (DeltaSI) was significantly higher in 10(6) labeled cells after 1-day culture than that in the same number of labeled cells after 8-day culture and that in 5 x 10(5) labeled cells, particularly on T(2)*WI (P<.05). Among pulse sequences, T(2)*WI demonstrated the highest DeltaSI (P<.05). CONCLUSION: The human umbilical cord blood MSCs can be labeled with Fe(2)O(3)-PLL without significant change in viability and apoptosis. The suspension of labeled MSCs can be imaged with standard 1.5-T MR equipment.

Cell Survival↗

Determination of biogenic amines in HeLa cell lysate by 6-oxy-(N-succinimidyl acetate)-9-(2'-methoxycarbonyl) fluorescein and micellar electrokinetic capillary chromatography with laser-induced fluorescence detection.

An MEKC-LIF method using 6-oxy-(N-succinimidyl acetate)-9-(2'-methoxy-carbonyl) fluorescein (SAMF) newly synthesized in our lab as a labeling reagent for the separation and determination of eight typical biogenic amines was proposed. After careful study of the derivatization condition such as pH value, reagent concentration, temperature, and reaction time, derivatization reaction was accomplished as quickly as 10 min with stable yield. Optimal separation of SAMF-labeled amines was achieved with a running buffer (pH 9.3) containing 30 mM boric acid, 25 mM SDS, and 20% v/v ACN. The proposed method allowed biogenic amines to be determined with LODs as low as 0.25-2.5 nmol/L and RSD values from 0.4 to 4.5%. The present method has been successfully used to monitor biogenic amines in HeLa cells and fish samples. This study exploits the potential of MEKC-LIF with SAMF labeling as a tool for monitoring biogenic amines involved in complex physiological and behavioral processes in various matrices.

Biogenic Amines↗

Bromophenols coupled with methyl gamma-ureidobutyrate and bromophenol sulfates from the red alga Rhodomela confervoides.

Four new bromophenols C-N coupled with methyl gamma-ureidobutyrate (1-4), a phenylethanol bromophenol (5), and three phenylethanol sulfate bromophenols (6-8) have been isolated from polar fractions of an ethanolic extract of the red alga Rhodomela confervoides. On the basis of spectroscopic evidence including HRMS and 2D NMR data, the structures of the new compounds were determined as methyl N'-(2,3-dibromo-4,5-dihydroxybenzyl)-gamma-ureidobutyrate (1), methyl N,N'-bis(2,3-dibromo-4,5-dihydroxybenzyl)-gamma-ureidobutyrate (2), methyl N'-[3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyl]-gamma-ureidobutyrate (3), methyl N'-(2,3-dibromo-4,5-dihydroxybenzyl)-N'-[3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyl]-gamma-ureidobutyrate (4), 2,3-dibromo-4,5-dihydroxyphenylethanol (5), 2,3-dibromo-4,5-dihydroxyphenylethanol sulfate (6), 3-bromo-4,5-dihydroxyphenylethanol sulfate (7), and 3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxyphenylethanol sulfate (8). The cytotoxicity of all compounds was evaluated against several human cancer cell lines including human colon cancer (HCT-8), hepatoma (Bel7402), stomach cancer (BGC-823), lung adenocarcinoma (A549), and human ovarian cancer (A2780). Among them, the phenylethanol and the phenylethanol sulfate bromophenols (5-8) showed moderate cytotoxicity against all tested cell lines.

Antineoplastic Agents↗

Study on a new precolumn derivatization method in the determination of metformin hydrochloride.

Metformin hydrochloride is successfully determined by reversed-phase high-performance liquid chromatography with a new precolumn derivatization method using 9,10-anthraquinone-2-sulfonyl chloride as the derivatization agent. Several derivatization systems are tried to optimize the derivatization conditions, and a new post-derivatization treatment method is established. The derivatization product is analyzed on a Lichrosper C18 column (6.0 mm x 150 mm, 5 microm) at 256 nm with methanol-water (70:30, v/v) as the mobile phase. The calibration curves of the derivatives for the UV detector (0.01-4 mg/L) are linear with respect to peak area. The detection limit (peak area) for the metformin hydrochloride is 0.01 mg/L for a signal-to-noise ratio of 3:1. In human plasma, the detection limit is 0.02 mg/L. This assay is rapid, sensitive, and highly reproducible.

Calibration↗

[Effects of nitric oxide synthases on kidney in early phase of endotoxic shock in macaque].

OBJECTIVE: To study the pathogenesis of kidney injury in early phase of endotoxic shock. METHODS: Eleven macaques were anesthetized and randomly divided into two groups: an experimental group (n=6), receiving a dose of 2.8 mg/kg lipopolysaccharides (LPS) by intravenous injection, and a control group (n=5), injected with normal saline of 1 ml/kg. The animals were sacrificed 120 minutes following endotoxin injection. With immunohistochemical technique endothelium nitric oxide synthases (eNOS) and inducible nitric oxide synthases (iNOS) protein of the renal tissue were determined. The ultrastructure was studied with electron microscope. Acid phosphatase (ACP) was also assayed. RESULTS: One hundred and twenty minutes after LPS challenge, damages to the glomerulus and renal tubules were found, and lysosomes were increased in the renal cells. iNOS was positively expressed in vascular endothelium of glomerulus and epithelium of renal tubules in the experimental group, but not in the control group. eNOS was positively expressed in vascular endothelium of glomerulus; and peri-vascular region in both groups. CONCLUSION: The present findings indicate that NO contributed to the pathogenesis of renal injury in the early phase of endotoxic shock in primates.

Animals↗

Simultaneous determination of water-soluble and fat-soluble synthetic colorants in foodstuff by high-performance liquid chromatography-diode array detection-electrospray mass spectrometry.

An accurate method was developed for the simultaneous determination of water-Tartrazine, Amaranth, Ponceau 4R, Sunset Yellow FCF, and fat-Sudan (I-IV), synthetic soluble colorants in foodstuff. This method uses dimethylsulfoxide (DMSO) as the extraction solvent in the sample preparation process and high performance liquid chromatography (HPLC)-diode array detector (DAD)-electrospray mass spectrometry (ESI-MS), applying selected ion recording in positive/negative alternate mode to acquire mass spectral data, as the analytical technique. Linearity of around three orders in the magnitude of concentration was generally obtained. Detection and quantification limits of the investigated dyes, which were evaluated at signal to noise ratio of 3 for detection limit and 10 for quantification limit, were in the ranges of 0.01-4 and 0.03-11.2 ng, respectively. The recoveries of the eight synthetic colorants in four matrices ranged from 93.2 to 108.3%. Relative standard deviations of less than 8.2% were also achieved. This method has been applied successfully in the determination of water-soluble colorants in the soft drink and the delicious ginger, and fat-soluble dyes in chilli powders and chilli spices.

Beverages↗

Ultrasound-assisted liquid-phase microextraction and high-performance liquid chromatographic determination of nitric oxide produced in PC12 cells using 1,3,5,7-tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacene.

To reveal the function of nitric oxide (NO) that has been reported as an intracellular second messenger and a diffusible intercellular messenger, a novel ultrasound-assisted liquid-phase microextraction (ULPME) and high-performance liquid chromatographic method for the determination of NO produced in PC12 cells has been developed. 1,3,5,7-Tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacene (DAMBO-CO(2)Et) has been used for NO trapping to form corresponding triazole (DAMBO-CO(2)Et-T). The enrichment factor of DAMBO-CO(2)Et-T reached 150 in 2.5 min with the proposed ULPME method. NO produced in (200+/-5) PC12 cells has been determined directly with the detection limit (S/N=3) of 2.5 x 10(-13)mol/L (5a mol), which was the lowest in HPLC determination of NO reported yet. The proposed method was sensitive, selective, convenient and rapid, which has great prospect in the qualitative and quantitative determination of NO in minimal biological samples.

Animals↗

Liquid-phase microextraction combined with high-performance liquid chromatography for the determination of local anaesthetics in human urine.

A simple liquid-phase microextraction (LPME) device combined with high-performance liquid chromatography (HPLC) is presented for the simultaneous analysis of local anaesthetics, lidocaine, bupivacaine, and tetracaine, from human urine sample. An organic solvent showed good compatibility with the mobile phase of the HPLC, o-dibutyl phthalate, was selected. Local anaesthetics are extracted from 6 ml of the feed aqueous solution and human urine sample into a water-immiscible organic solvent suspended at the needle tip of the microsyringe, then the organic solvent was directly introduced to a reversed-phase HPLC system. The kind of the organic extraction solvent, the stirring rate, the pH value of the aqueous feed solution, and the extraction time have been discussed. Under the optimized extraction conditions, high enrichment factors (more than 86.0-fold) and significant sample clean-up for all of studied local anaesthetics were achieved within 30 min. The detection limits (lower than 0.05 microg/ml) were comparable with previously reported gas chromatography methods. This method was applied to specimen of patient who was treated with extradural anaesthesia of lidocaine, bupivacaine, and tetracaine, and revealed that simultaneous determination of above three local anaesthetics in human urine was possible.

Anesthetics, Local↗

[Expression of melanoma antigen gene in urinary transitional cell carcinomas].

OBJECTIVE: To evaluate the significance of melanoma antigen (MAGE) gene expression in bladder transitional cell carcinoma (TCC). METHODS: MAGE-A1, A2, A3, A4 mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR) in 3 clusters bladder TCC cells and 20 samples of bladder TCC patients (T(1) 7 samples, T(2) 5 samples, T(3) 6 samples, T(4) 2 samples, G(1) 1 samples, G(2) 11 samples, G(3) 8 samples). MAGE-A4 protein was detected by immunohistochemistry in 105 samples of bladder TCC patients (T(1) 35 samples, T(2) 12 samples, T(3) 26 samples, T(4) 13 samples, G(1) 13 samples, G(2) 44 samples, G(3) 48 samples). RESULTS: Three clusters bladder TCC cells had MAGE gene mRNA expression. In detection of MAGE mRNA of 20 samples of bladder TCC patients, 12 samples (60%) expressed MAGE-A1, 16 samples (80%) expressed MAGE-A2, 11 samples (55%) expressed MAGE-A3, 18 samples (90%) expressed MAGE-A4, 8 samples (40%) expressed all of MAGE-A1--A4. In 105 bladder TCC samples, 53 samples (50%) expressed MAGE-A4 protein. Strong expression (++ or +++) was significant higher in higher grade (13 samples/48 samples) or stage (14 samples/51 samples) than in lower grade (2 samples/57 samples) or stage (0 samples/35 samples). CONCLUSION: MAGE gene highly expresses in bladder TCC. Bladder TCC of high grade or high stage has higher MAGE-A4 protein strong expression.

Adult↗

[Expression of SSX2 gene in human urologic neoplasms].

OBJECTIVE: To investigate the expression of SSX(2)gene in human renal cell carcinoma and urinary transitional cell carcinoma. METHODS: Reverse-transcription polymerase chain reaction (RT-PCR) was used for detecting SSX(2) gene in the specimens from renal cell carcinoma (n = 26), urinary transitional cell carcinoma (n = 27) and in 15 specimens taken from the tumor surrounding tissues. RESULTS: Positive expression of SSX(2) gene at mRNA was detected in 69% renal cell carcinomas (18/26), in 81% urinary transitional cell carcinomas (22/27). The mRNA of SSX(2) was not detected in the 15 specimens from tumor surrounding tissues. CONCLUSION: The SSX(2) gene is highly expressed in human renal cell carcinoma and urinary transitional cell carcinoma.

Adult↗

Prevention of core cell damage in isolated islets of Langerhans by low temperature preconditioning.

AIM: To study the core cell damage in isolated islets of Langerhans and its prevention by low temperature preconditioning (26 degrees). METHODS: Islets were cultured at 37 degrees for 7-14 d after isolation, and then at 26 degrees for 2, 4 and 7 d before additional culture at 37 degrees for another 7 d. Core cell damage in the isolated islets was monitored by video-microscopy and analyzed quantitatively by use of a computer-assisted image analysis system. The analysis included daily measurement of the diameter and the area of the isolated islets and the area of the core cell damage that developed in those islets over time during culture. Histology and TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay were used to characterize the cell damage and to monitor islet function. RESULTS: Microscopic analysis showed that during the 7 to 14 d of culture at 37 degrees, core cell damage occurred in the larger islets with diameters >200 microm, which included both necrotic and apoptotic cell death. Low temperature (26 degrees) culture could prevent core cell damage of isolated islets. The 7-d culture procedure at 26 degrees could inhibit most of the core cell (excluding diameters >300 microm) damages when the islets were re-warmed at 37 degrees. CONCLUSION: Our results indicate that core cell damage within isolated islets of Langerhans correlates with the size of islets. Low temperature (26 degrees) culture can prevent core cell damage in isolated islets, and successfully precondition these islets for incubation at 37 degrees. These novel findings may help to understand the pathophysiology of early loss of islet tissue after transplantation, and may provide a new strategy to improve graft function in the clinical setting of islet transplantation.

Animals↗

Bromophenols coupled with derivatives of amino acids and nucleosides from the red alga Rhodomela confervoides.

Three new bromophenols coupled with pyroglutamic acid derivatives and one bromophenol coupled with deoxyguanosine were obtained from the red alga Rhodomela confervoides. By spectroscopic methods including 2D NMR and single-crystal X-ray structure analysis their structures were elucidated as N-(2,3-dibromo-4,5-dihydroxybenzyl)methyl pyroglutamate (1), N-(2,3-dibromo-4,5-dihydroxybenzyl)pyroglutamic acid (2), N-[3-bromo-2-(2,3-dibromo-4,5-dihydroxybenzyl)-4,5-dihydroxybenzyl]methyl pyroglutamate (3), and 2-N-(2,3-dibromo-4,5-dihydroxybenzylamino)deoxyguanosine (4), respectively. Compounds 1-4 were evaluated against several microorganisms and human cancer cell lines, but found inactive. To our knowledge this is the first report of bromophenols coupled with amino acid or nucleoside derivatives through the C-N bond.

Amino Acids↗