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Biomedical subjects

M Zhu

Publications and source records attributed to M Zhu.

At least 91 records · Page 5Linked to original sources

[A study on transmission of pathogenic bacteria of rampant caries from mothers to children].

OBJECTIVE: In order to prevent rampant caries in children, the transmission from mothers to children of pathogenic bacteria of rampant caries will be investigated in this study. METHODS: By using fingerprint technique, the genetic diversities of Streptococcus mutans(S. mutans) were examined respectively in 10 children with rampant caries, 10 children without rampant caries S. mutans, 10 2-year-old caries-free children, and their mothers. The levels of S. mutans in saliva of mothers were also examined. For fingerprinting, chromosomal DNA of isolates obtained from plaques of these children and from saliva of their mothers were digested with restriction endonuclease HindIII and examined by electrophoresis on 0.7% agarose gel. DNA fingerprints were obtained and analyzed for genotypic similarities. RESULTS: The DNA fingerprint patterns showed that the proportion of children whose genotypes of S. mutans matched their mothers in the children with rampant caries was 70%, which is similar to that in the children without rampant caries and the caries-free children (60% respectively). The number of distinct genotypes of S. mutans harbored in children with rampant caries was, on average, greater than the number of genotypes present in children without rampant caries and caries-free children (1.7, 1.2 and 1.1 respectively, P < 0.05). Mothers of children with rampant caries had similar levels of S. mutans in saliva as those mothers of children without rampant caries and caries-free children. However, mothers of children with rampant caries had significantly higher DMFT scores and higher levels of Streptococcus sobrinus (S. sobrinus) in saliva than the mothers of caries-free children did. CONCLUSION: In all child groups, mothers were probably the main source of infection with S. mutans. Rampant caries in children is associated with DMFT scores and levels of S. sobrinus in saliva of mothers. In the initiation of rampant caries experienced by a child, the transmission of pathogenic bacteria from mothers to children may be the main source.

Adult↗

[Analysis on 16S rDNA sequence of rhizobia isolated from Kummerowla sp].

Based on the previous studies on numerical taxonomy, SDS-PAGE of whole-cell protein and DNA hybridization, the rhizobial strains isolated from Kummerowia sp. in semi-arid area of North-west constituted a new subgroup, the 16S rDNA sequence of representative strain SH714 were tested. The unrooted phylogenetic tree was produced. In this tree, the strain SH714 with Sinorhizobium xinjiangensis, S. fredii, S. meliloti, S. medicae, S. saheli and S. teranga constituted a branch of Sinorhizobium. Within this branch, the similarity valuse of 16S rDNA sequence between strain SH714 and S. xinjiangesis, S. fredii, S. meliloti, S. medicae, S. saheli and S. teranga were 97.4%, 97.5%, 96.8%, 96.7%, 97.2% and 95.6% respectively, the values were more than 95%, this indicated that these known species should belong to the same genus. The values of DNA homology between type strains of these species were less than 70%. Thus, the strain SH714 represented a new rhizobial species, and there were some diversity between SH714 and known rhizobial species in phenotypic feature and composition of protein.

DNA, Bacterial↗

[Inhibition of cultured human retinal pigment epithelial cell by tissue-type plasminogen activator].

PURPOSE: To study the inhibition effects of human retinal pigment epithelial (RPE) cells by tissue-type plasminogen activator(tPA). METHODS: The effects of tPA on human RPE cells proliferation were studied by viable cell counting and methyl thiazolyl tetrazolium (MTT) colorimetric assay. Flow cytometry (FCM) analysis was used to examine RPE cell cycles. RESULTS: The proliferation of human RPE cells could be inhibited by 0.1-3 micrograms/ml tPA (P < 0.05). The effects of inhibition were depended on the drug density and the action time directly (P < 0.01). 5 micrograms/ml tPA was deadly-toxic to human RPE cells. FCM revealed that the cells in S phase increased 9.8% (P < 0.05) and cells in G2M phase decreased 6.6% (P < 0.05), compared to controls. CONCLUSION: tPA at a certain dose range and period could inhibit human RPE cells proliferation without deadly-toxicity. The human RPE cells growth could be interfered in S phase by tPA.

Cell Division↗

[SFE-CGC determination of tanshinone in Salvia miltiorrhiza Bunge].

AIM: To analyze tanshinone IIA in Salvia miltiorrhiza Bunge by supercritical fluid extraction (SFE) coupled with capillary gas chromatography (CGC). METHODS: The solution pressure was predicted by solubility parameter theory, the extraction conditions were confirmed by orthogonal design in which temperature, volume of modifier and dynamic extracting volume had been studied in three levels. RESULTS: This method is quick, convenient, precise and accurate, added sample recovery is 95.3%, RSD = 4.27%. CONCLUSION: Compared with the method of ultrasonic extraction, significantly better qualitative and quantitative result could be obtained by SFE-CGC and expend less organic solvent then make less pollution.

Abietanes↗

[A primary study on the relationship between condyle position and anterior distraction for cross bite caused by cleft lip and palate].

OBJECTIVE: To evaluate the changes of the position of the mandibular condyle in cleft lip and palate patients with secondary bony cross-bite deformity after early orthodontic treatment with extraoral bow and facial mask. METHODS: The TMJ topograph 6 months before and after treatment was investigated in 8 patients. The width of the anterior,superior and posterior space of the TMJ was compared prior to and post treatment. RESULTS: The results showed that there was no significant difference in the width of the three spaces of the TMJ before and after treatment. CONCLUSION: The position of the mandibular condyle had no significant change after maxillary anterior distraction with extraoral bow and facial mask.

English Abstract↗

[Soft tissue profile changes after maxillary protraction in complete unilateral cleft lip and palate patients].

OBJECTIVE: The purpose is to evaluate the soft tissue profile changes in patients with complete unilateral cleft lip and palate treated after maxillary protraction. METHODS: 12 patients with complete unilateral cleft lip and palate were treated using maxillary protraction. Lateral cephalometric films were taken before and after treatment. Soft tissue profile changes were evaluated by comparing the difference of soft tissue landmarks between two films. RESULTS: After maxillary protraction, the convex of nose (S-Ns-PRN) increased significantly (P<0.001), the convex of maxillary soft tissue was increased by 3.8 degree (P<0.01). The convex of middle face changed significantly (P<0.01). In mandible angle S-Ns-SM was decreased one degree (P>0.05). Vertically, there was no significant difference in upper face height (P>0.05). Total face height increased significantly (P<0.01). CONCLUSION: After treatment of maxillary protraction, the soft tissue profile in patients with complete cleft lip and palate improved significantly.

English Abstract↗

Development of a green fluorescent protein microplate assay for the screening of chemopreventive agents.

Here we develop a rapid, cell-based, functional assay to screen and identify naturally occurring or synthetic chemicals with chemopreventive activity. We constructed a reporter gene that consists of the gene-encoding green fluorescent protein (GFP) under the transcriptional control of the thymidine kinase (TK) promoter adjacent to which concatamerized EpRE regulatory elements were inserted. Human hepatoma HepG2 cells were transfected with the EpRE/TK-GFP reporter plasmid, and clones with low GFP background expression and high tBHQ-induced GFP expression were isolated. These GFP reporter cells were seeded into a 96-well microtiter plate, incubated for 24 h, and then treated with test compounds for an additional 24 h. The GFP level and DNA content (as an internal cell survival control) of cells in the 96-well plate were measured subsequently using a fluorescence plate reader. Known inducers of phase II enzymes, such as tert-butylhydroquinone, beta-naphthoflavone, and sulforaphane, significantly increased the GFP level in the HepG2 reporter cells. In an initial screening of a chemical library, we identified a synthetic compound whose inducing ability significantly exceeds (1.6-fold) that of the best currently known phase II enzyme inducers. The experimental results indicate that this cell system makes possible a new high throughput screening approach to identify novel chemopreventive molecules.

Antineoplastic Agents↗

Presence of oxidized cholesterol in caveolae uncouples active platelet-derived growth factor receptors from tyrosine kinase substrates.

Platelet-derived growth factor receptor beta (PDGFRbeta) in fibroblasts is concentrated in caveolae where it controls the tyrosine phosphorylation of multiple proteins. Caveolae are enriched in cholesterol and sphingolipids, but the role of these lipids in PDGFR signal transduction is unknown. We report that introduction of cholest-4-en-3-one into caveolae membranes uncouples PDGFR autophosphorylation from tyrosine phosphorylation of neighboring proteins. Cholest-4-en-3-one appears to interfere with the normal interaction between PDGFR and its partners. The results suggest that tightly packed caveolae lipids form a membrane platform that functions as a lipid scaffold for organizing the molecular interactions of multiple signaling pathways.

Caveolae↗

Association of Grb2, Gads, and phospholipase C-gamma 1 with phosphorylated LAT tyrosine residues. Effect of LAT tyrosine mutations on T cell angigen receptor-mediated signaling.

The linker for activation of T cells (LAT) is a critical adaptor molecule required for T cell antigen receptor (TCR)-mediated signaling and thymocyte development. Upon T cell activation, LAT becomes highly phosphorylated on tyrosine residues, and Grb2, Gads, and phospholipase C (PLC)-gamma1 bind LAT via Src homology-2 domains. In LAT-deficient mutant Jurkat cells, TCR engagement fails to induce ERK activation, Ca(2+) flux, and activation of AP-1 and NF-AT. We mapped the tyrosine residues in LAT responsible for interaction with these specific signaling molecules by expressing LAT mutants with tyrosine to phenylalanine mutations in LAT-deficient cells. Our results showed that three distal tyrosines, Tyr(171), Tyr(191), and Tyr(226), are responsible for Grb2-binding; Tyr(171), and Tyr(191), but not Tyr(226), are necessary for Gads binding. Mutation of Tyr(132) alone abolished PLC-gamma1 binding. Mutation of all three distal tyrosines also abolished PLC-gamma1 binding, suggesting there might be multiple binding sites for PLC-gamma1. Mutation of Tyr(132) affected calcium flux and blocked Erk and NF-AT activation. Since Grb2 binding is not affected by this mutation, these results strongly suggest that PLC-gamma activation regulates Ras activation in these cells. Mutation of individual Grb2 binding sites had no functional effect, but mutation of two or three of these sites, in combination, also affected Erk and NF-AT activation.

Adaptor Proteins, Signal Transducing↗

Quantitation of tyrosine hydroxylase protein in the locus coeruleus from postmortem human brain.

In this study, we developed an immuno-autoradiographic method to obtain quantitative estimates of tyrosine hydroxylase (TH) protein in tissue sections from post-mortem human brain. Protein from tissue sections containing the locus coeruleus (LC) was directly transferred to a polyvinylidene fluoride (PVDF) membrane. Immunoreactive TH on PVDF membranes was identified with optimized concentrations of TH antibody followed by application of [125I]labeled secondary antibody. Quantities of TH on autoradiograms were estimated by comparing optical densities of transferred immunoblots to a calibrated standard curve produced with purified recombinant TH dotted onto the same PVDF membranes. Amounts of TH-immunoreactivity in the LC were proportional to the thickness of tissue sections up to 15 micrometer. However, the amounts of total protein, as measured by Ponceau S staining, were linearly related to section thicknesses up to 30 micrometer. Comparisons of quantities of immunoreactive TH in the LC using this method to amounts determined using traditional Western blotting, in which LC tissue was punched from adjacent sections from the same subject, showed a positive correlation (r(2)=0.99, P<0.01). Using the transfer immunoblot method, an uneven distribution of TH protein was observed along the rostrocaudal axis of the human LC (P<0.01). This method may provide a sensitive and useful tool for the study of the role of human TH expression in the pathophysiology of psychiatric disease.

Autoradiography↗

[Cloning and analysis of different regions of SRY promoter].

The luciferase systems were used to assay the promoter activity of SRY gene with clones of different parts of the 5' flanking region within 544 bp which has basal promoter activity. The results were that the 179 bp region from nt. -353 to nt. -174 upstream of the first ATG included a silent element; the 49 bp region from nt. -112 to nt. -63 included an enhanced element and the 63 bp region from nt. -174 to nt. -112 included an essential promoter sequency for gene transcription. These results give some important clues to elucidate the expression and regulation mechanisms of SRY gene.

5' Flanking Region↗

A recombinant antibody-targeted plasminogen activator with high affinity for activated platelets increases thrombolytic potency in vitro and in vivo.

To increase thrombolytic specificity of urokinase (uPA), we engineered a recombinant chimeric plasminogen activator SZ51Hu-scuPA, which consists of a humanized monoclonal antibody (SZ-51Hu) specifically against P-selectin on activated human platelet and a single-chain urokinase (scuPA). The cDNA, encoding scuPA amino acids 1-411, was inserted in 5' end to 3' end orientation immediately after the CH3 of SZ-51Hu heavy-chain sequence in the expression vector alphaLys30. The resulting construct alphaLys30-SZ51VH/Hu-scuPA was used to transfect into SP2/0 murine myeloma cell line, which was pretransfected with SZ51Hu light chain. The fusion protein SZ51Hu-scuPA was expressed at 5 mg/L in the supernatant of cell culture. The fusion protein purified by affinity chromatography had a molecular weight of 160 kDa with fibrinolytic activity of 39,000 IU/mg and its affinity to activated human platelet was 67% of the parent murine mAb SZ-51. The thrombolytic property of the fusion protein was first characterized in an in vitro system, which consists of a 125I-fibrin-labeled human plasma clot containing different concentrations of human platelets suspended in citrated human plasma. Fifty percent lysis was reached with SZ51Hu-scuPA in 1 hour at a concentration of 20 IU/mL or in 2 hours at a concentration of 10 IU/ mL, which was much faster than uPA at the same concentration. The maximal lysis of the clots by SZ51Hu-scuPA was 4.1 to 8.4 times more potent than that by uPA. The fusion protein was further characterized in the hamster pulmonary embolism model with clots prepared from fresh platelet-rich human plasma containing 125I-labeled fibrinogen. The thrombolytic activity of SZ51-scuPA was 3.9 times more potent than that of uPA at 2,000 IU/kg in this model. Almost no significant fibrinogen breakdown was observed either in vitro and in vivo.

Animals↗

MR imaging of pituitary morphology in idiopathic intracranial hypertension.

The aim of this study was to investigate the morphologic changes of the pituitary gland in patients with the clinical diagnosis of idiopathic intracranial hypertension (IIH). Qualitative and quantitative analyses of pituitary morphology were performed in normal subjects (n = 23), patients with the clinical diagnosis of IIH (n = 40), and patients with acute increased intracranial pressure (AICP; n = 37) caused by acute head trauma. The loss of pituitary height (concavity) on the sagittal T1-weighted image was classified into five categories: I = normal, II = superior concavity that was mild (<(1/3) the height of the sella), III = moderate (between (1/3) and (2/3) concavity of height of sella), IV = severe (>(2/3) concavity of height of sella), and V = empty sella. The area ratio of pituitary gland to sella turcica measured in the midsagittal plane was quantified. Clinical records were retrospectively reviewed to correlate with magnetic resonance (MR) findings. Using moderate concavity (>(1/3)) as the minimum criterion for abnormality, IIH patients had an 85% incidence of morphologic changes with 80% sensitivity and 92% specificity. Empty sella (almost complete concavity of the sella) was found in only 2.5% of patients with IIH. Quantitative analysis of the pituitary gland/sella turcica area ratio showed a significant decrease in patients with IIH (P < 0.0001) but no significant difference between the normal subjects and AICP patients. A posterior deviation of the pituitary stalk was seen in 43% of patients. No enlargement of the ventricles or sulcal effacement was seen in IIH patients. Routine brain MR examination of patients with IIH frequently shows morphologic changes of the pituitary gland ranging from various degrees of concavity to (rarely) the extreme case of an empty sella. The etiology is unknown and may be related to the severity and duration of elevated CSF pressure. Such findings may be useful to facilitate the diagnosis of IIH, particularly in patients with equivocal clinical findings or when IIH is not suspected. J. Magn. Reson. Imaging 2000;12:808-813.

Acute Disease↗

The human hyaloid system: cell death and vascular regression.

The present study had investigated the roles of apoptosis and necrosis in the regression of the human fetal hyaloid vasculature. Normal human fetal hyaloid specimens (n = 67) ranging from 10 to 20 weeks' gestation were studied. Specimens were either immunolabeled with anti-von Willebrand factor and major histocompatibility complex class I antibodies or investigated using the terminal-deoxyribonucleotidyl transferase-mediated dUTP-biotin DNA nick-end labeling technique. A fluorescent DNA-binding dye acridine orange/ethidium bromide mixture was also applied to unfixed flat mounts of hyaloid vasculature and some specimens were processed for transmission electron microscopy. Vascular regression including cell loss in the connecting vessels, stretching and thinning of the vasa hyaloidea propria, tunica vasculosa lentis and the pupillary membrane was clearly evident after 13 weeks' gestation. Cresyl violet staining revealed condensed cells and pyknotic bodies throughout the hyaloid system; cell death occurred either in single cells or along small capillary segments associated with vascular regression. Acridine orange/ethidium bromide staining showed DNA condensation at early and late stages of cell death. Similarly, DNA nick-end labeling was positive in endothelial cells, pericytes and vessel and non-vessel associated hyalocytes. The observation of hyalocytes juxtaposed to cytolysed endothelial cells may indicate a role for these cells in vascular regression. Features of apoptosis were more evident during early vascular regression whilst necrosis was increasingly evident at later stages.

Apoptosis↗

Expression of angiotensin AT(1) and AT(2) receptors in adult rat cardiomyocytes after myocardial infarction. A single-cell reverse transcriptase-polymerase chain reaction study.

The effector hormone of the renin-angiotensin system, angiotensin II, plays a major role in cardiovascular regulation. In rats, both angiotensin receptor subtypes, AT(1) and AT(2), are up-regulated after myocardial infarction but previous studies failed to identify the cell types which express the AT(2) receptor in the heart. To address this question we established a single-cell reverse transcriptase-polymerase chain reaction for AT(1) and AT(2) receptors to determine whether these receptor subtypes are expressed in adult rat cardiomyocytes before and 1 day after myocardial infarction. By laser-assisted cell picking, section profiles of single cells without genomic DNA contamination were isolated. After dividing samples into two identical aliquots, polymerase chain reaction amplification for AT(1) and AT(2) receptors was carried out and polymerase chain reaction products were subjected to gel electrophoresis. Compared to control (n = 4) and sham-operated animals (n = 4), the number of cardiomyocytes expressing the AT(1) receptor mRNA 1 day after myocardial infarction (n = 4) was not changed (42% and 33% versus 45%, respectively). On the other hand, AT(2) receptor mRNA was expressed in 8% and 13%, respectively, of cardiomyocytes gained from control (n = 4) and sham-operated animals (n = 4) and in 14% isolated after myocardial infarction (n = 4). These results demonstrate for the first time that the AT(2) receptor is expressed in adult cardiomyocytes in vivo. They further suggest that the previously observed up-regulation of cardiac AT(1) and AT(2) receptors after myocardial infarction involves cell types other than cardiomyocytes.

Animals↗

Closed-loop control of muscle length through motor unit recruitment in load-moving conditions.

Neuroprostheses aimed at restoring lost movement in the limbs of spinal cord injured individuals are being developed in this laboratory. As part of this program, we have designed a digital proportional-integral-derivative controller integrated with a stimulation system which effects recruitment of motor units according to the size principle. This system is intended to control muscle length while shortening against fixed loads. Feline sciatic nerves were exposed and stimulated with ramp, triangular, sinusoidal, staircase and random signals as test inputs. Changes in muscle length and effective time delay under different conditions were measured and analyzed. Differences of tracking quality between open- and closed-loop conditions were examined through analysis of variance as well as the differences between small (250g) and large (1kg) loads. The results showed that parameters used to compare muscle length output to the input signals were dramatically improved in the closed-loop trials as compared to the open-loop condition. Mean squared correlation coefficients between input and output signals for ramp signals increased by 0.019, and for triangular signals by 0.12. Mean peak cross correlation between input and output signals for sinusoidal waveforms increased by 0.06, with decreases in time to peak cross correlation (effective time delay) from 195 to 38ms. In slow random signals (power up to 0.5Hz), peak cross correlation went from 0.74 to 0.89, and time-to-peak cross correlation decreased from 205 to 55ms. In fast random signals (power up to 1Hz), peak cross correlation went from 0.82 to 0.89, and time-to-peak cross correlation from 200 to 65ms. For staircase signals, both rise times and mean steady-state errors decreased. It was found that, once the length range was set, the load weight had no effect on tracking performance. Analysis of mean square error demonstrated that for all signals tested, the feedback decreased the tracking error significantly, whereas, again, load had no effect. The results suggest that tracking is vastly improved by using a closed-loop system to control muscle length, and that load does not affect the quality of signal tracking as measured by standard control system analysis methods.

Animals↗

Biexponential recovery model of lumbar viscoelastic laxity and reflexive muscular activity after prolonged cyclic loading.

OBJECTIVES: To determine the rest duration required for full recovery of reflexive muscular activity and laxity/creep induced in the lumbar viscoelastic structures (e.g., ligaments, discs, etc.) after 50 min of cyclic loading, and to develop a model describing such recovery. BACKGROUND: It is well established that steady, cyclic or vibratory loading of the lumbar spine induces laxity/creep in its viscoelastic structures. It was also shown that such viscoelastic creep does not fully recover when subjected to rest equal in duration to the loading period. Rest periods of 24 h, however, were more than sufficient to allow full recovery. The exact period of time allowing full recovery of viscoelastic laxity/creep, and its pattern is not known. It is also not known what is the duration required for full recovery of reflexive muscular activity lost due to the laxity/creep induced in the spine during cyclic loading. METHODS: The lumbar spine of 'in vivo' feline preparations was subjected to 50 min of 0.25 Hz cyclic loading applied v ia the L4/5 supraspinal ligament. At the end of the loading period the spine was subjected to prolonged rest, interrupted by a single cycle loading applied hourly for measurement purposes until the laxity was fully recovered (>90%). Reflexive EMG activity was recorded with wire electrodes from the L-1-L-7 multifidus muscles. A biexponential model was fitted to the load and EMG recorded in the recovery period in order to represent viscous and elastic components of structures with different architecture (e.g., disc vs. ligament). RESULTS: Full recovery of the laxity induced by 50 min of cyclic loading at 0.25 Hz required 7 h and was successfully fitted with a biexponential model. Similarly, EMG activity was fully recovered in 4 hours, and often exceeded its initial value during the following 3 h. CONCLUSIONS: Full recovery of laxity induced in the lumbar viscoelastic structures by a given period of cyclic loading requires rest periods, which are several folds longer than the loading duration. Similarly, reflexive muscular activity requires 4 h of rest in order to be restored. Meanwhile, significant laxity can be present in the joints, exposing the spine to potential injury and low back pain. Increased EMG activity at the end of the recovery period may indicate that pain was possibly induced in the spinal structures, inducing hyperexcitability of the muscles during passive loading. RELEVANCE: Although the data was derived from a feline model, and its extrapolation to the human model is not straightforward, the general pattern of decreasing reflexive muscular activity with cyclic loading is expected in both species. Therefore, workers who subject their spine to periods of cyclic loading may be exposed to prolonged periods of laxity beyond the neutral zone limits, without protection from the muscles and therefore the risk of possible injury and low back pain. Pain and muscle hyperexcitability could also be a factor associated with cyclic loading, being expressed several hours after work was completed.

Animals↗