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Biomedical subjects

Z Tang

Publications and source records attributed to Z Tang.

At least 235 records · Page 13Linked to original sources

A high-sensitivity CCD system for parallel electron energy-loss spectroscopy (CCD for EELS).

A cooled frame transfer CCD camera system was developed and tested as a parallel detector in an electron energy-loss spectrometer mounted on a transmission electron microscope. The use of a shutterless camera with a frame transfer CCD collected virtually 100% of the photon signal with a reasonably fast acquisition time. The system detective quantum efficiency was over 90% under normal experimental conditions. Because of the low channel to channel gain variations in the CCD, the signal-to-noise ratio and the detection limit were substantially better than that obtained with a silicon intensified target (SIT) camera, and direct fitting to the standard data was feasible. Quantitation at the phosphorus L edge generated from a phosphoprotein, phosvitin, showed that, under identical experimental conditions, direct fitting of spectra obtained with this CCD system gave better sensitivity than that given by the SIT camera system. Because of its larger pixel charge well, the CCD system can also operate at a much higher beam current, resulting in a significant reduction in the time required for elemental mapping at a given sensitivity.

Image Processing, Computer-Assisted↗

The GATA-4 transcription factor transactivates the cardiac muscle-specific troponin C promoter-enhancer in nonmuscle cells.

The unique contractile phenotype of cardiac myocytes is determined by the expression of a set of cardiac muscle-specific genes. By analogy to other mammalian developmental systems, it is likely that the coordinate expression of cardiac genes is controlled by lineage-specific transcription factors that interact with promoter and enhancer elements in the transcriptional regulatory regions of these genes. Although previous reports have identified several cardiac muscle-specific transcriptional elements, relatively little is known about the lineage-specific transcription factors that regulate these elements. In this report, we demonstrate that the slow/cardiac muscle-specific troponin C (cTnC) enhancer contains a specific binding site for the lineage-restricted zinc finger transcription factor GATA-4. This GATA-4-binding site is required for enhancer activity in primary cardiac myocytes. Moreover, the cTnC enhancer can be transactivated by overexpression of GATA-4 in non-cardiac muscle cells such as NIH 3T3 cells. In situ hybridization studies demonstrate that GATA-4 and cTnC have overlapping patterns of expression in the hearts of postimplantation mouse embryos and that GATA-4 gene expression precedes cTnC expression. Indirect immunofluorescence reveals GATA-4 expression in cultured cardiac myocytes from neonatal rats. Taken together, these results are consistent with a model in which GATA-4 functions to direct tissue-specific gene expression during mammalian cardiac development.

3T3 Cells↗

Negative regulation of the wee1 protein kinase by direct action of the nim1/cdr1 mitotic inducer.

The wee1 protein kinase suppresses the entry into mitosis by mediating the inhibitory tyrosine phosphorylation of p34cdc2. Genetic studies have suggested that the nim1 protein kinase (also known as cdr1) acts as a positive regulator of mitosis by down-regulating the wee1 pathway in yeast cells. We have overexpressed the nim1 protein in both bacteria and insect cells. The recombinant nim1 protein autophosphorylates on both tyrosine and serine residues and can phosphorylate the isolated wee1 protein directly in a cell-free system. The nim1-catalyzed phosphorylation of the wee1 protein occurs in its C-terminal region and leads to a substantial drop in its activity as a cdc2-specific tyrosine kinase. This nim1-dependent inhibition of the wee1 protein kinase can be reversed readily in vitro by treatment with a protein phosphatase. These experiments provide direct biochemical evidence that the wee1 protein is subject to negative regulation by phosphorylation and indicate that the nim1 protein acts as an inhibitory, wee1-specific kinase.

Animals↗

Two distinct mechanisms for negative regulation of the Wee1 protein kinase.

The Wee1 protein kinase negatively regulates the entry into mitosis by catalyzing the inhibitory tyrosine phosphorylation of the Cdc2 protein. To examine the potential mechanisms for Wee1 regulation during the cell cycle, we have introduced a recombinant form of the fission yeast Wee1 protein kinase into Xenopus egg extracts. We find that the Wee1 protein undergoes dramatic changes in its phosphorylation state and kinase activity during the cell cycle. The Wee1 protein oscillates between an underphosphorylated 107 kDa form during interphase and a hyperphosphorylated 170 kDa version at mitosis. The mitosis-specific hyperphosphorylation of the Wee1 protein results in a substantial reduction in its activity as a Cdc2-specific tyrosine kinase. This phosphorylation occurs in the N-terminal region of the protein that lies outside the C-terminal catalytic domain, which was recently shown to be a substrate for the fission yeast Nim1 protein kinase. These experiments demonstrate the existence of a Wee1 regulatory system, consisting of both a Wee1-inhibitory kinase and a Wee1-stimulatory phosphatase, which controls the phosphorylation of the N-terminal region of the Wee1 protein. Moreover, these findings indicate that there are apparently two potential mechanisms for negative regulation of the Wee1 protein, one involving phosphorylation of its C-terminal domain by the Nim1 protein and the other involving phosphorylation of its N-terminal region by a different kinase.

Amino Acid Sequence↗

Signal transducing molecules and glycosyl-phosphatidylinositol-linked proteins form a caveolin-rich insoluble complex in MDCK cells.

GPI-linked protein molecules become Triton-insoluble during polarized sorting to the apical cell surface of epithelial cells. These insoluble complexes, enriched in cholesterol, glycolipids, and GPI-linked proteins, have been isolated by flotation on sucrose density gradients and are thought to contain the putative GPI-sorting machinery. As the cellular origin and molecular protein components of this complex remain unknown, we have begun to characterize these low-density insoluble complexes isolated from MDCK cells. We find that these complexes, which represent 0.4-0.8% of the plasma membrane, ultrastructurally resemble caveolae and are over 150-fold enriched in a model GPI-anchored protein and caveolin, a caveolar marker protein. However, they exclude many other plasma membrane associated molecules and organelle-specific marker enzymes, suggesting that they represent microdomains of the plasma membrane. In addition to caveolin, these insoluble complexes contain a subset of hydrophobic plasma membrane proteins and cytoplasmically-oriented signaling molecules, including: (a) GTP-binding proteins--both small and heterotrimeric; (b) annex II--an apical calcium-regulated phospholipid binding protein with a demonstrated role in exocytic fusion events; (c) c-Yes--an apically localized member of the Src family of non-receptor type protein-tyrosine kinases; and (d) an unidentified serine-kinase activity. As we demonstrate that caveolin is both a transmembrane molecule and a major phospho-acceptor component of these complexes, we propose that caveolin could function as a transmembrane adaptor molecule that couples luminal GPI-linked proteins with cytoplasmically oriented signaling molecules during GPI-membrane trafficking or GPI-mediated signal transduction events. In addition, our results have implications for understanding v-Src transformation and the actions of cholera and pertussis toxins on hetero-trimeric G proteins.

Animals↗

[Expression of DNA fragment encoding for immunodominant epitopes of 150kd phosphoprotein of human cytomegalovirus in Escherichia coli].

HVYP fragment, which contains the DNA encoding region for immunodominant epitopes of 150 kd phosphoprotein (pp150) of human cytomegalovirus, was isolated from recombinant plasmid pHCY1 and the inserted into the PstI site of E. coli expression vector pUC8. The resulted recombinant plasmid pUHVYP1 with correct orientation and in-frame was transformed into E. coli JM107. Induced by isopropyl-beta-D-thiogalactoside (IPTG), the transformants produced HVYP fusion protein with molecular weight of about 44-45 kd, as determined by 12% SDS-PAGE. These results from Western blot and ELISA showed that the expressed protein could be recognized by the specific anti-HCMV IgG and IgM antibodies of patients' serum. MAC-ELISA further proved that the fusion protein was a potential antigen for diagnostic reagents.

Cloning, Molecular↗

[The changes in free radical and prostaglandin metabolism in newborn infants with asphyxia].

Blood plasma thromboxane-B2 (TXB2) and 6-keto-PGF1 alpha levels were determined by radioimmunoassay technique during the first six to sixteen hours of life in 16 newborn infants with severe asphyxia, 18 newborn infants with mild asphyxia and 27 normal term neonates. Plasma lipid peroxidation (LPO) content was measured by TBA-colour-contrast method in 15 infants with severe asphyxia, 17 infants with mild asphyxia and 24 healthy term newborn infants. The results showed that blood plasma LPO, TXB2 and 6-keto-PGF1 alpha levels in infants suffering from severe asphyxia were higher than those in infants with mild asphyxia and normal infants (P < 0.01), but no significant difference was noted between the mild asphyxia group and normal control group (P > 0.01). These suggest that production of free radicals is increased and prostaglandin metabolism is triggered in the infants with severe asphyxia, that cerebral ischemia and hypoxia caused by asphyxia contributes to the augmented production of prostaglandins and free radicals, and that accumulation of free radicals and prostaglandin enhances brain damage and the metabolism of arachidonic acid appears to be an important source of the free radicals in infants with intrauterine asphyxia.

6-Ketoprostaglandin F1 alpha↗

An experimental study on acupuncture anti-hemorrhagic shock.

The effect of acupuncture at Neiguan (P 6) on regulating hemorrhagic shock in 90 rabbits was observed. All the results suggested that acupuncture at Neiguan could raise blood pressure, protect cardiac pump function, correct the disturbance of secretion and metabolism of humoral factors, reduce blood adhesion to normal level and thus achieve anti-hemorrhagic shock. This study has provided scientific basis for expounding the mechanism of acupuncture in anti-shock.

Acupuncture Therapy↗

[The influence of neonatal hyperbilirubinemia on neonatal behavior].

Two groups of neonates, 30 cases with hyperbilirubinemia (307.8 +/- 99.86 mumol/L) and 30 normal newborns (control group with bilirubin 80.37 +/- 35.74 mumol/L), were evaluated with Neonatal Behavioral Assessment Scale (NBAS) and followed up at the age of 2 months and 5 months. All cases in the two groups were evaluated with NBAS at ages of 2-3 days, 12-14 days and 26-28 days. The results showed that there were statistically significant differences between the two groups in the aspects of attention and social responsiveness, the ability to modulate state of consciousness, the motor integrity and the reflexes. We also found that while the level of bilirubin was below 205.2 mumol/L, there was no correlation between the level of bilirubin and NBAS (r = -0.095, P > 0.02), but when bilirubin was at or above the level of 205.2 mumol/L, there was significant negative correlation (r = -0.53, P < 0.001) between the two. The follow-up revealed that 9 of 23 jaundiced infants had abnormal auditory reactions or abnormal muscle tonus at the age of 2 months; 8 of the 9 infants recovered, but 1 still had abnormal muscle tonus by the age of 5 months. The study suggests that hyperbilirubinemia above the level of 205.2 mumol/L would affect neonatal behavior and the greater the severity of jaundice, the more significant the influence on neonatal behavior. The effects on some jaundiced infants may last 5 months or more. NBAS is a valuable scale for early detection of mild brain dysfunctions which are less obvious on clinical examination.

Brain↗

[Construction of clone about major DNA fragment coding for 150kd phosphoprotein of human cytomegalovirus and its restriction site analysis].

Human cytomegalovirus (HCMV) DNA EcoRI-Y fragment, which is major DNA fragment coding for 150kd phosphoprotein (pp150) of HCMV, was separated and recovered from recombinant cosmid pCM1015. The recombinant plasmids pHCY1 and pHCY2, which contain major DNA fragment coding for pp150, were constructed successfully, using the plasmid pUC19 as vector. The recombinant plasmids were analysed by restriction endonucleases and determined by Southern hybridization. The restriction sites of DNA encoding region for immunodominant epitopes of HCMV pp150 were analysed by GENEPRO system. The region could be digested by 76 kinds of restriction endonucleases. The sizes of cleaved fragments were between 4-2667 bp.

Cloning, Molecular↗

Radioiodinated anti-hepatocellular carcinoma (HCC) ferritin. Targeting therapy, tumor imaging and anti-antibody response in HCC patients with hepatic arterial infusion.

Radioimmunoimaging and radioimmunotherapy with radioiodinated anti-(hepatocellular carcinoma ferritin) antibody (131I- or 125I-FtAb) have been applied in patients with primary liver cancer. A total of 41 patients with surgically unresectable hepatocellular carcinoma (HCC) and receiving hepatic artery ligation and cannulation during exploratory laparotomy were treated with this regimen by intrahepatic arterial infusion. Compared with the control group, a decline of serum alpha-fetoprotein (65.7% versus 42.9%) and shrinkage of tumor (68.3% versus 33.9%) were observed in the treated group, and a higher second-look resection rate (31.7% versus 5.1%) and longer survival (1-year: 61.0% versus 37.3%, 3-year: 25.0% versus 6.9%) resulted. The administration of antibody through a hepatic arterial catheter (n = 16) was compared with intravenous injection (n = 17) in terms of the tumor-imaging sensitivity in 33 patients with liver cancer. The results indicated that hepatic arterial infusion was superior to intravenous injection. The sensitivity 7 days after the administration was 100% in the i.a. group and 76.5% in the i.v. group, the uptake ratio of tumor to liver being 1.74 +/- 0.57 in the former and 1.34 +/- 0.29 in the latter. Furthermore, intrahepatic arterial infusion revealed a lower anti-antibody detection rate than intravenous injection (0/14 versus 4/11).

Antibody Formation↗

[Preliminary use of recombinant glycoprotein 52kd as an antigen in the diagnosis of human cytomegalovirus infection].

Interest in the human cytomegalovirus (HCMV) mainly derives from its associations with congenital malformations, mental retardation, and severe or fatal infections in immunosuppressed individuals such as transplant patients, tumor and AIDS patients. It is evidenced that there has been a need for a rapid and sensitive methods to detect an ongoing acute infection. The recent studies showed that high titers of antibody to the glycoprotein 52kd are present in sera of patients undergoing acute HCMV infection. However, purification of individual glycoprotein from HCMV-infected cells is a daunting prospect. HCMV glycoprotein 52 kd expressed via recombinant DNA techniques are a promising approach to solve this problem. In order to evaluate the diagnostic value of the recombinant glycoprotein 52 kd antigenic code region for HCMV infection, we have used the polymerase chain reaction (PCR) and recombinant DNA techniques to construct successfully the high-level expression plasmid pHCMV containing the HCMV GP-52 kd antigenic code region, with the predicted protein at levels up to 20% in total bacterial protein. The expressed protein was purified from SDS-PAGE, used as an antigen in Western-blot, and reacted with 12 cases of the positive sera, 4 cases of the negative sera, following by reaction with HRP-labelled horse IgG antibody against human. The results indicated that the approach we are using to detect antibody to HCMV acute infection are as sensitive as general serological methods such as ELISA, with the advantages of easy preparation of antigen with high quantity, and clinical practicability.

Antibodies, Viral↗

[Influence of moxibustion on TXA2 and PGI2 in plasma of rat infected epidemic hemorrhagic fever virus (EHFV)].

In this paper, the model of rat infected EHFV was made and the influence of moxibustion on TXA2 and PGI2 in its plasma was observed. The results show that the content of TXA2 increased and PGI2 decreased in rat significantly after the abdominal inoculation of EHFV, and the content of TXA2 decreased and PGI2 increased markedly in rat infected EHFV to normal level after treatment with moxibustion, suggesting that the regulative function of moxibustion on TXA2 and PGI2 is one of its nerve-endocrine-immune regulations to the body, and there is an important significance. This study provides an important reference for mechanism exploration of moxibustion preventing and treating EHF.

6-Ketoprostaglandin F1 alpha↗

Clinical evaluation of 99mTc-MIBI myocardial tomography for detecting coronary artery disease.

To evaluate the clinical efficacy of 99mTc-MIBI myocardial tomography for detecting coronary artery disease, 34 consecutive patients who underwent both 99mTc-MIBI study and coronary arteriography were included in the present study. Ten without and 24 with significant coronary artery disease were documented by coronary arteriography. The overall sensitivity and specificity of 99mTc-MIBI study for detecting coronary artery disease were 91.8% (22/24) and 80.0% (8/10), respectively. In subset studies, 16 of 17 (94.1%) patients with previous myocardial infarction and 6 of 7 (85.7%) patients with significant CAD but without MI were identified as abnormal. Reversible perfusion abnormalities were shown in 4 of 5 patients undergoing both exercise and rest studies. Rest perfusion abnormalities were shown in 2 patients with unstable angina pectoris who underwent only rest study. In conclusion, 99mTc-MIBI myocardial tomography provides a reliable method for detecting coronary artery disease. Rest study has high sensitivity for identifying patients with MI, and exercise/rest study can identify patients with significant CAD but without MI.

Coronary Disease↗

[Analysis of genetic models and gene effects on main agronomy characters in rapeseed].

According to four different genetic models, the genetic patterns of 8 agronomy traits were analysed by using the data of 24 generations which included positive and negative cross of 81008 x Tower, both of the varieties are of good quality. The results showed that none of 8 characters could fit in with additive-dominance models. Epistasis was found in all of these characters, and it has significant effect on generation means. Seed weight/plant and some other main yield characters are controlled by duplicate interaction genes. The interaction between triple genes or multiple genes needs to be utilized in yield heterosis.

Brassica↗

Prothrombin expression in the adult and fetal rabbit liver.

Plasma prothrombin levels in newborn humans are lower than in adults. The same is true of many newborn and fetal mammals, including the rabbit. To determine if the lower levels are due to less expression of the protein, we have compared mRNA for prothrombin in fetal and adult rabbit liver. Northern blots were hybridized with a cDNA for rabbit prothrombin revealing a single mRNA of approximately 2 kb in both adult and fetal animals. mRNA specific for prothrombin was quantitated by slot blotting of RNA prepared from adults and fetuses aged 21 d to term (31 d). Prothrombin-specific mRNA in fetuses was greater than 50% of that in adults even when the fetal plasma prothrombin was only 15% of the adult level. This suggests that low plasma levels in the fetuses are not the result of less transcription. Examination of liver sections revealed that the predominant tissue in the fetus is hematopoietic, not hepatic. In the youngest fetuses, less than 20% of the liver consisted of hepatocytes, yet these fetuses expressed more than 50% of the adult level of prothrombin-specific mRNA. Thus, transcription of prothrombin mRNA may be proceeding at a greater rate in the fetal hepatocyte than in the adult, or hematopoietic cells may be expressing the protein. We conclude that in fetal rabbit liver, prothrombin is expressed at a high level relative to the hepatocyte content and that the cause of the low plasma levels is posttranscriptional.

Animals↗

Hb Westmead: an alpha 2-globin gene mutation detected by polymerase chain reaction and Stu I cleavage.

Hb Westmead or alpha 2 122(H5)His----Gln beta 2 is one of the most common hemoglobin variants in Guangxi, a province in Southern China. The alpha 2-globin DNA sequence of a carrier for this variant was selectively amplified by the polymerase chain reaction and analyzed with the restriction enzyme Stu I. We found that this mutant globin was encoded at the alpha 2 locus and that the CAC----CAG mutation at codon 122 created a new Stu I restriction site. It is easy to detect this mutation by these methods.

Base Sequence↗