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Biomedical subjects

M Zhang

Publications and source records attributed to M Zhang.

At least 55 records · Page 3Linked to original sources

Effects on protein and mRNA expression levels of p53 induced by fluoride in human embryonic hepatocytes.

We investigated the effects of protein and mRNA expression levels on p53 induced by fluoride in human embryo hepatocyte L-02 cells. The protein and mRNA levels of p53 in L-02 cells were measured after in vitro cultured L-02 was exposed to sodium fluoride at different doses (40, 80, and 160 microg/ml) for 24 h. The results showed that the cell survival rate of L-02 cells in the high dose fluoride group was significantly lower than that of the control group. The protein expression levels of p53 in the middle and high dose fluoride group were significantly higher than in the control group and elevated with increasing fluoride concentration. The mRNA expression levels of p53 in the fluoride groups were markedly higher than in the control group. The mRNA expression level of p53 in the high dose fluoride group was however lower compared to the middle dose fluoride group, but similar to the low dose fluoride group. These finding suggest that fluoride can decrease the L-02 cells survival rate and induce protein and mRNA expressions of p53; however, there is no consistency between the protein expression level of p53 and the mRNA expression level.

Blotting, Western↗

Glial cell line-derived neurotrophic factor-induced signaling in Schwann cells.

Glial cell line-derived neurotrophic factor (GDNF), a known survival factor for neurons, has recently been shown to stimulate the migration of Schwann cells (SCs) and to enhance myelination. GDNF exerts its biological effects by activating the Ret tyrosine kinase in the presence of glycosylphosphatidylinositol-linked receptor, GDNF family receptor (GFR) alpha1. In Ret-negative cells, the alternative transmembrane coreceptor is the 140-kDa isoform of neural cell adhesion molecule (NCAM) associated with a non-receptor tyrosine kinase Fyn. We confirmed that GDNF, GFRalpha1 and NCAM are expressed in neonatal rat SCs. We found that GDNF induces an increase in the partitioning of NCAM and heparan sulfate proteoglycan agrin into lipid rafts and that heparinase inhibits GDNF-signaling in SCs. In addition to activation of extracellular signal-regulated kinases, and phosphorylation of cAMP response element binding protein, we found that cAMP-dependent protein kinase A and protein kinase C are involved in GDNF-mediated signaling in SCs. Although GDNF did not promote the differentiation of purified SCs into the myelinating phenotype, it enhanced myelination in neuron-SC cocultures. We conclude that GDNF utilizes NCAM signaling pathways to regulate SC function prior to myelination and at early stages of myelin formation.

Agrin↗

In vitro response of tumour cells to non-uniform irradiation.

This study examines differences in tumour cellular response using clonogenic cell survival between uniform and non-uniform irradiation. Cells were irradiated with a 6 MV x-ray intensity-modulated beam, in a single large flask (i.e. intercellular communication is possible) or in three small flasks (i.e. intercellular communication is inhibited across the dose gradient). For non-small-cell lung cancer and melanoma cell lines, the dose response over the entire cell culture was significantly different between freely communicating cell cultures and those with inhibited communication across the dose non-uniformity. Communicating cells exhibited poorer survival in the low dose region of the field but improved survival in the high dose region. In general, the response to non-uniform irradiation appeared to 'average out' over the entire cell culture. This was not seen when intercellular communication was inhibited. The results add strength to the body of evidence regarding bystander effects and the inter-dependence of cellular response.

Carcinoma, Large Cell↗

Dissociation of electrolytes in a nano-aqueous system within single-wall carbon nanotubes.

Research on material incorporation within single-wall carbon nanotubes (SWNTs) through aqueous solutions of various electrolytes is performed for the purpose of providing a foundation for future application of SWNTs to, for example, drug delivery systems. We have determined that the optical spectra of SWNTs were significantly affected when SWNTs that had opened holes or removed caps were treated through immersion in an aqueous solution of electrolytes, followed by drying at room temperature; however, the spectra of SWNTs without opened holes or removed caps were not subjected to such treatment. We infer that when the sucked solutions remained inside the tubes, even after drying (the nano-aqueous system), the electrolyte was dissociated into ions, which was likely to change the electronic states of SWNTs. On the other hand, when the SWNTs were well-dried under vacuum, no remarkable changes in their optical spectra were observed.

Letter↗

Effect of resveratrol on peritoneal macrophages in rats with severe acute pancreatitis.

OBJECTIVE: The literature on resveratrol in severe acute pancreatitis (SAP) is limited though it has been widely studied in infections and trauma. The aim of this study was to investigate the inhibitory effect of resveratrol on inflammatory responses in a rat model of SAP. METHODS: Male Sprague-Dawley (SD) rats were randomly divided into 3 groups: SAP group, resveratrol group and control group. 4.0% sodium taurocholate was injected into the pancreatic duct to induce SAP. In the resveratrol group, resveratrol (10 mg/kg) was injected through penal vein 5 min after SAP was induced. The peritoneal macrophages of the rats were collected 3, 6 and 12 h after stimulus and then incubated for 24 h. The expression of nuclear factor kappa B (NF-kappaB) and inducible nitric oxide synthase (iNOS) in peritoneal macrophages was measured. The levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1) and nitric oxide (NO) in culture medium of peritoneal macrophages and serum of rats were evaluated. RESULTS: Histological examination of pancreas indicated that the damage in the SAP group was more severe than that in the resveratrol group. The expression of NF-kappaB and iNOS in peritoneal macrophages was significantly higher in the SAP group than in the resveratrol group. The concentrations of TNF-alpha, IL-1 and NO in culture medium and serum were significantly elevated in the SAP group when compared with the resveratrol group. CONCLUSIONS: The inhibiting effect on the inflammatory response and the decreased expression of TNF-alpha, IL-1 and NO in peritoneal macrophages suggest resveratrol as a novel anti-inflammatory agent for reducing the severity of SAP.

Acute Disease↗

Interactions between charged residues in the transmembrane segments of the voltage-sensing domain in the hERG channel.

Studies on voltage-gated K channels such as Shaker have shown that positive charges in the voltage-sensor (S4) can form salt bridges with negative charges in the surrounding transmembrane segments in a state-dependent manner, and different charge pairings can stabilize the channels in closed or open states. The goal of this study is to identify such charge interactions in the hERG channel. This knowledge can provide constraints on the spatial relationship among transmembrane segments in the channel's voltage-sensing domain, which are necessary for modeling its structure. We first study the effects of reversing S4's positive charges on channel activation. Reversing positive charges at the outer (K525D) and inner (K538D) ends of S4 markedly accelerates hERG activation, whereas reversing the 4 positive charges in between either has no effect or slows activation. We then use the 'mutant cycle analysis' to test whether D456 (outer end of S2) and D411 (inner end of S1) can pair with K525 and K538, respectively. Other positive charges predicted to be able, or unable, to interact with D456 or D411 are also included in the analysis. The results are consistent with predictions based on the distribution of these charged residues, and confirm that there is functional coupling between D456 and K525 and between D411 and K538.

Amino Acid Sequence↗

Double-antigen sandwich ELISA for detection of antibodies to SARS-associated coronavirus in human serum.

The study presented here was conducted to evaluate the performance of a double-antigen sandwich ELISA to detect antibodies in human serum against the coronavirus associated with severe acute respiratory syndrome (SARS). A recombinant partial nucleocapsid protein of SARS-associated coronavirus was used as a serodiagnostic antigen in the ELISA. A total of 2892 clinical serum samples were tested with the ELISA kit, which positively identified 25 of 35 (71.4%) samples of patients with confirmed SARS infection, 286 of 407 (70%) samples of patients suspected of having SARS, 229 of 302 (75.8%) samples of convalescent SARS patients, and 0 of 544 samples obtained from healthcare workers; only 1 of 1604 clinical samples obtained from patients with other diseases demonstrated a weakly positive result. These results indicate that the double-antigen sandwich ELISA is an effective screening method for the serodiagnosis of SARS-associated coronavirus.

Antibodies, Viral↗

Cardiolipin in energy transducing membranes.

Cardiolipin is a phospholipid located exclusively in energy transducing membranes such as the bacterial cytoplasmic membrane and the inner membrane of mitochondria. It plays both a structural and a functional role in many multimeric complexes associated with these membranes. The role of cardiolipin in higher order organization of components of the mitochondrial respiratory chain revealed by a combined molecular genetic and biochemical approach is described.

Animals↗

Diagnosis of human cytomegalovirus intrauterine infection using fetal cells from maternal blood.

OBJECTIVE: The sensitivity and specificity for the noninvasive prenatal diagnosis of human cytomegalovirus intrauterine infection were estimated by using isolating single fetal cells from maternal peripheral blood. METHODS: Micromanipulation techniques were employed to isolate single fetal nucleated erythroblasts from 273 maternal blood samples. SRY gene and HCMV-DNA in single fetal cells were detected by multiple primed in situ labeling (PRINS) from 76 HCMV-DNA positive samples of maternal peripheral blood. 273 samples of maternal peripheral blood were tested for SRY gene and HCMV-DNA in single fetal cells by primed extension preamplification (PEP) and polymerase chain reaction (PCR). RESULTS: The detection rate of fetal cells from maternal blood was 100% with micromanipulation techniques. The sensitivity of PRINS for SRY gene detection was 97.56% and its specificity was 100%. The sensitivity and specificity of PEP and PCR for SRY gene detection were 97.39% and 99.17%, respectively. The sensitivity of PRINS for HCMV-DNA detection was 92.68% and the specificity was 100%. The sensitivity and specificity of PEP and PCR for HCMV-DNA detection were 95.12%and 100%, respectively. CONCLUSION: The technique for noninvasive prenatal detection of intrauterine infection of HCMV using single fetal cells from maternal peripheral blood by using PRINS and PEP and PCR is more reliable than the CMV-DNA detection in peripheral maternal blood, amniocentesis or percutaneous umbilical blood sampling.

Cytomegalovirus↗

Expression of endothelial nitric oxide synthase traffic inducer in the placentas of women with pre-eclampsia.

OBJECTIVE: To investigate the expression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) in the placenta of women with pre-eclampsia (PE) and discuss its role in the pathogenesis of PE. METHODS: Western blot analysis was used to detect the expression of NOSTRIN and endothelial nitric oxide synthase (eNOS). The activity of eNOS in placental tissue was assayed by spectrophotometry. Serum and placental NO(2)/NO(3), the stable metabolic end product of NO, was measured using nitrate reductase. RESULTS: Western blot analysis showed that the protein level of NOSTRIN was significantly higher in women with PE than in the control group (P<0.01). However, no significant difference between groups was observed in the expression of placental eNOS (P>0.05). The activity of eNOS was significantly decreased in women with PE (13.727+/-3.58 U/mg) compared with the control group (21.69+/-3.84 U/mg) (P<0.01). The placental levels of NO(2)/NO(3) were significantly lower in women with PE (27.53+/-8.51 micromol/mg) than in the control group (44.38+/-9.59 micromol/mg) (P<0.01). The levels of serum NO(2)/NO(3) were significantly lower in women with PE (48.56+/-8.49 micromol/L) than in the control group (65.37+/-9.61 micromol/L) (P<0.01). A significant negative correlation existed between the expression of NOSTRIN protein and the activity of eNOS in the placental tissues of women with PE (r=-0.57, P<0.01). CONCLUSION: The level of NOSTRIN expression in the placental tissues of women with PE is increased and is negatively correlated with the activity of eNOS-which may play an important role in the pathogenesis of PE.

Adaptor Proteins, Signal Transducing↗

A novel DNA vaccine based on ubiquitin-proteasome pathway targeting 'self'-antigens expressed in melanoma/melanocyte.

Cancer vaccine that targets 'self'-antigens expressed at high levels in tumor cells is a potentially useful immunotherapy, but immunological tolerance often defeats this strategy. Here, we describe the use of a naked DNA vaccine encoding a self tumor antigen, tyrosinase-related protein 2, to whose N-terminus ubiquitin is fused in a 'nonremovable' fashion. Unlike conventional DNA vaccines, this vaccine broke the tolerance and induced protective immunity to melanoma in C57BL/6 mice, as evaluated by tumor growth, survival rate and lung metastasis. The protective immunity was cancelled in the proteasome activator PA28alpha/beta knockout mice. Moreover, this vaccination exhibited therapeutic effects on melanoma implanted before vaccination. Our findings provide evidence for the first time that naked DNA vaccines encoding a ubiquitin-fused self-antigen preferentially induce the main effector CD8+ T cells through efficient proteolysis mediated by the ubiquitin-proteasome pathway, and lead the way to strategies aimed at targeting tissue differentiation antigens expressed by tumors.

Animals↗

Thiopurine methyltransferase and 6-thioguanine nucleotide measurement: early experience of use in clinical practice.

BACKGROUND: Azathioprine and 6-mercaptopurine (6-MP) are well established for the treatment of inflammatory bowel disease (IBD). Assessing thiopurine methyltransferase (TPMT) status has been recommended to reduce the risk of serious toxicity. Measuring red blood cell (RBC) 6-thioguanine nucleotide (6-TGN) concentrations has been recommended for dose adjustment. AIM: To describe the results of measuring TPMT activity and genotype, and 6-TGN concentration in New Zealand. METHODS: Canterbury Health Laboratories provided these analyses for New Zealand. Those with low TPMT activity also underwent genotyping. All results were collated and analysed descriptively. 6-TGN concentrations were correlated with the dose of thiopurine when known. RESULTS: TPMT enzyme activity (range 1-22 U/mL) from 574 patients showed a trimodal distribution. Genotyping results matched this distribution with only mild overlap between (*1/*1) homozygote and (*1/*3) heterozygote groups. One patient without TPMT measurement before therapy had life-threatening neutropenia and was later found to have (*3/*3) genotype. TPMT analysis probably prevented two further such cases. Of 884 6-TGN concentrations (range 0-1434 pmol/10(8) RBC), 41, 39 and 20% were within, below, and above the therapeutic range of 235-450 pmol/10(8) RBC, respectively. Leucopenia was seen in some patients with high 6-TGN. 6-MMP concentrations in 177 patients with low 6-TGN suggested non-compliance in 31, underdosing in 130, and preferential metabolism of 6-MP to 6-methylmercaptopurine in 16. There was poor correlation between azathioprine dose and 6-TGN concentration (r(2) = 0.002), supporting 6-TGN monitoring. CONCLUSIONS: Measurement of TPMT enzyme activity and 6-TGN concentration has been well-integrated into clinical practice. These tests should reduce the risk of toxicity and improve efficacy with thiopurines in patients with IBD.

Azathioprine↗

The high prognostic value of the histologic grade at the deep invasive front of tongue squamous cell carcinoma.

BACKGROUND: Although many histopathologic factors in squamous cell carcinoma of the tongue predict the prognosis, the major predictive factors have not been identified clearly. This study analyzed the prognostic value of the histologic grade at the deep invasive front of tongue squamous cell carcinoma. METHODS: The clinicopathologic features of 124 consecutive patients seen between January 1985 and December 1999 with previously untreated squamous cell carcinoma of the tongue were reviewed. Their mean age was 58.5 years (range: 23-90) and the male-female ratio was 1.8: 1 (79 men and 45 women). There were 41, 40, 30, and 13 cases at stage I to stage IV, respectively. The clinicopathologic factors, especially the histologic grade at the deep invasive front (invasive front grade, IFG), were analyzed to determine factors predicting prognosis. RESULTS: The 5-year disease-free survival rate of the patients treated with curative aim only was 66.7%. Clinicopathologic factors significantly associated with the prognosis were T classification, tumor size, stage classification, tumor depth, macroscopic appearance, cervical lymph node metastasis (nodal metastasis), microvascular invasion, and IFG. In a multivariate analysis, patients with tumor depth >/=4 mm, IFG >/=8 points, and nodal metastasis had a reduced disease-free survival and IFG >/=11 points had a predictive value for nodal metastasis (odds ratio: 7.34; P = 0.0019). CONCLUSION: This study found that a high IFG malignancy score had a high prognostic value for squamous cell carcinoma of the tongue.

Adult↗

Age- and sex-specific body composition of Chinese children.

UNLABELLED: We examined age- and sex-specific body compositions of Chinese children by the bioelectrical impedance method. The subjects were a total of 587 children aged 6-14 y who had normal relative weight. In all ages, boys had larger fat-free mass and lower percent body fat (%BF) than girls did. Even in the subjects with BMI <20 kg/m2, more than one quarter of them had high %BF. CONCLUSION: Chinese children may have higher %BF than that predicted by BMI.

Adolescent↗

A quantitative food frequency questionnaire for women in southeast China: development and reproducibility.

This study describes the development and reproducibility of a 128-item quantitative food frequency questionnaire (FFQ) to measure usual food consumption for women in southeast China. The FFQ was pre-tested using 51 Chinese women who recently migrated to Australia. Cronbach's alpha coefficient was 0.81 for internal consistency. The reliability of the FFQ was then assessed by another test-retest study. A sample of 41 women residing in southeast China was interviewed twice within 12 weeks. Intraclass correlation coefficients were moderate to high for mean food group consumption (0.43-0.96) and mean daily nutrient intakes (0.47-0.89). Kappa statistics for eating habits ranged from 0.27 to 0.89 in the test-retest. The mean ratio of energy intake to basal metabolic rate was 1.73 (S.D. 0.39) in both test and retest samples. The study confirmed that the FFQ method using standard containers is appropriate to assess dietary intake for women in southeast China.

Adult↗

Atrial natriuretic peptide inhibits the actions of FSH and forskolin in meiotic maturation of pig oocytes via different signalling pathways.

Atrial natriuretic peptide (ANP) as well as its receptors is found in mammalian ovary and follicular cells and its function in oocyte meiotic maturation has also been reported in Xenopus, hamster and rat. But the results are controversial and the physiological mechanism of ANP on oocyte maturation is not clear, especially the relationship between gonadotrophin and ANP as well as the signal transduction, and these need further study. The present study conducted experiments to examine these questions by using drug treatment and Western blot analysis and focused on pig oocyte meiotic maturation and cumulus expansion in vitro. The results revealed that ANP could inhibited FSH-induced pig oocyte maturation and cumulus expansion and prevent the full phosphorylation of mitogen-activated protein kinase in both oocytes and cumulus cells, and that these inhibitory effects could be mimicked by 8-Br-cyclic guanosine 5'-monophosphate (8-Br-cGMP), but blocked by a protein kinase G (PKG) inhibitor KT5823. Zaprinast, a cGMP-specific phosphodiesterase inhibitor, could enhance the inhibitory effect of ANP on oocyte maturation. A specific analogue of ANP, C-ANP-(4-23), which binds to the natriuretic peptide receptor-C (NPRC), had no effect in either FSH-induced or spontaneous oocyte maturation. Treatment with forskolin, a stimulator of adenylate cyclase, had a biphasic effect; 44 h treatment induced cumulus expansion but inhibited oocyte maturation while 2 h treatment induced maturation of cumulus-enclosed oocytes (CEOs). Both ANP and C-ANP-(4-23) could inhibit the effect of forskolin on CEO maturation, and these inhibitory effects of ANP/C-ANP-(4-23) could be blocked by preincubation with pertussis toxin (PT), consistent with mediation by a Gi protein(s) in the cumulus cells. All these results suggest that ANP is a multifunctional regulator of FSH and forskolin on pig CEO maturation by two signalling mechanisms: one is via a cGMP/PKG pathway, the other is via NPRC receptors in cumulus cells and the activation of the PT-sensitive Gi protein(s).

Animals↗

Eruption sequestrum: x-ray microanalysis and microscopic findings.

A 12 year-old girl with eruption sequestrum of the mandibular left second molars is reported. Intraoral findings revealed that a small hard tissue fragment white in color and with bone-like hardness on the occlusal surface of the mandibular second molar which was erupting. Histopathologically, the fragments consisted of necrotized cortical bone. Chronic inflammatory alterations were also observed in the gingiva in the area of contact with the osseous tissue. X-ray microanalyzer findings revealed the percentages of calcium and phosphorous (by weight) as 78.41% and 21.59%, respectively, for a calcium to phosphorous ratio of 3.63, which was higher than that seen in normal osseous tissue.

Bone and Bones↗