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Biomedical subjects

J Q Zhu

Publications and source records attributed to J Q Zhu.

At least 19 recordsLinked to original sources

Biochemical and biophysical analysis of heptad repeat regions from the fusion protein of Menangle virus, a newly emergent paramyxovirus.

Menangle virus is a novel paramyxovirus isolated in Australia in 1997, but its classification position has not yet been finally settled. Here by using a computational program, LearnCoil-VMF, we determined the heptad repeat (HR) regions (HR1 and HR2) of Menangle virus F protein. Subsequently the HR1 and HR2 peptides were expressed as a single chain (named 2-Helix) connected by a six amino-acid linker as a GST fusion protein with an E. coli in vitro expression system. The GST-removed purified 2-Helix protein could form a stable trimer in vitro judging by gel-filtration and chemical cross-linking. CD spectra showed that the 2-Helix protein had a high percentage of alpha-helix and was very thermo-stable. Crystals of the 2-Helix protein preparations have been obtained in many conditions with hanging-drop diffusion method. These results indicated that Menangle virus has the common features of the fusion protein for other paramyxoviruses and should adopt a similar fusion mechanism to other members. As the HR regions of Menangle virus F protein could form stable six-helix bundle coiled coil structure, they should be used as drug target for the design of fusion inhibitors, as successfully used for other parmyxoviruses. This is especially relevant to such a newly emergent virus with zoonotic potentials.

Amino Acid Sequence↗

Organization of human ACAT-2 gene and its cell-type-specific promoter activity.

Acyl-CoA:cholesterol acyltransferase (ACAT) plays important roles in cellular cholesterol homeostasis. Two ACAT genes exist in mammals. We report here the genomic organization of human ACAT-2 gene and analysis of its promoter activity in various cell lines. The human ACAT-2 gene spans over 18 kb and contains 15 exons. Three transcription start sites and one poly(A) site are identified by the 5'/3'-RACE. In addition, the human ACAT-2 gene is linked to the insulin-like growth factor binding protein 6 (IGFBP-6) gene in a head-to-tail manner with a small intergenic region of about 1.2 kb. The 5'-flanking region of human ACAT-2 gene contains many potential cis-acting elements for multiple transcriptional regulatory factors but lacks TATA and CCAAT boxes. Using promoter-luciferase reporter assays, we demonstrate the transcriptional activity of ACAT-2 gene promoter is high in Caco-2 cells, especially after these cells become postconfluent and behave as intestinal enterocytes.

Base Sequence↗

[Ethanol-induced gastric mucosal injury and the protection of taurine against the injury in rats].

40 SD rats were randomly divided into control group, ethanol-induced injury group and taurine protective group for the measurement of the contents of endothelin (ET), nitric oxide synthetase (NOS), somatostatin (SS) and vasoactive intestinal peptide (VIP). The results are as follows. The gastric mucosa was damaged by 75% ethanol, being aggravated with prolonged application time, as shown by a significant increase of the ET content and decreases of the contents of NOS, VIP and SS in the gastric mucosa. In the taurine group, decrease of ET content and increase of the contents of NOS, SS and VIP in the gastric mucosa were observed. Apparently, the changes in ET, NOS, SS and VIP contents of gastric mucosa serve as reliable indices in the pathogenesis by ethanol and protective effect by taurine.

Animals↗

Contribution of a voltage-sensitive calcium release mechanism to contraction in cardiac ventricular myocytes.

The contribution of a voltage-sensitive release mechanism (VSRM) for sarcoplasmic reticulum (SR) Ca2+ to contraction was investigated in voltage-clamped ventricular myocytes at 37 degrees C. Na+ current was blocked with lidocaine. The VSRM exhibited steady-state inactivation (half-inactivation voltage: -47.6 mV; slope factor: 4.37 mV). When the VSRM was inactivated, contraction-voltage relationships were proportional to L-type Ca2+ current (ICa-L). When the VSRM was available, the relationship was sigmoidal, with contractions independent of voltage positive to -20 mV. VSRM and ICa-L contractions could be separated by activation-inactivation properties. VSRM contractions were extremely sensitive to ryanodine, thapsigargin, and conditioning protocols to reduce SR Ca2+ load. ICa-L contractions were less sensitive. When both VSRM and ICa-L were available, sigmoidal contraction-voltage relationships became bell-shaped with protocols to reduce SR Ca2+ load. Myocytes demonstrated restitution of contraction that was slower than restitution of ICa-L. Restitution was a property of the VSRM. Thus activation and recovery of the VSRM are important in coupling cardiac contraction to membrane potential, SR Ca2+ load, and activation interval.

Animals↗

Two-year follow-up of directly-observed intermittent regimens for smear-positive pulmonary tuberculosis in China.

SETTING: The tuberculosis component of the Infectious and Endemic Disease Control Project in the People's Republic of China is the largest single tuberculosis control project in the world using directly-observed therapy and standardized intermittent regimens. OBJECTIVE: To determine the two-year relapse and mortality rates following completion of treatment. DESIGN: A prospective cohort study of 649 cases cured in this project. The 306 new and 343 retreatment cases were treated under field conditions with 2H3R3Z3S3/4H3R3 and 2H3R3Z3E3S3/6H3R3E3, respectively. Following treatment completion, two sputum samples were collected every six months for two years and examined for acid-fast bacilli. Causes of death were identified. RESULTS: The two-year relapse rates for new and retreatment cases were 3.3% and 5.6%, respectively. Retreatment cases with delayed sputum conversion had a greater risk for subsequent relapse. The two-year mortality rate for new and retreatment cases was 3.3% and 8.5%, respectively. The higher mortality rate in retreatment cases was not attributable to relapse of disease, but rather to non-infectious sequelae of tuberculosis. CONCLUSION: The use of directly-observed intermittent regimens is effective in permanently removing infectious tuberculosis cases from the community.

Antitubercular Agents↗

"Voltage-activated Ca release" in rabbit, rat and guinea-pig cardiac myocytes, and modulation by internal cAMP.

It is widely believed that Ca release from the sarcoplasmic reticulum (SR) in heart muscle is due to "Ca-induced Ca-release" (CICR), triggered by transmembrane Ca entry. However, in intact guinea-pig cells or cells dialysed with cAMP there may be an additional mechanism - SR release may be activated directly by membrane depolarisation without Ca entry. The first objective of the present study was to investigate whether this "voltage-activated Ca release" (VACR) mechanism is present across species such as rabbit, rat and guinea-pig. The second objective was to characterise the dependence of a VACR mechanism on internal [cAMP]. Membrane current was measured with the whole-cell patch-clamp technique, intracellular [Ca] was monitored with Fura-2 (or a combination of Fluo-3/SNARF-1). Rapid changes of superfusate (within 100 ms) were made using a system which maintained cell temperature at 37 degrees C. We used a train of conditioning pulses to ensure a standard SR load before each test pulse. In rabbit myocytes dialysed with 100 microM cAMP, 89.6 +/- 7.0% of the control intracellular Ca (Cai) transient was still elicited by depolarisation during a switch to 5 mM Ni, which blocked pathways for Ca entry. This suggested that rabbit myocytes possess a VACR mechanism. The percentage of control Cai transient elicited by depolarisation in the presence of 5 mM Ni (i.e. magnitude of VACR) increased in a graded fashion with the pipette [cAMP] between zero and 100 microM. In rat myocytes dialysed with 50 microM cAMP, 64.4 +/- 6.2% of SR release was activated by depolarisation in the presence of 5 mM Ni, suggesting the presence of a VACR mechanism. The extent to which VACR triggered SR release increased with the pipette [cAMP] between zero and 50 microM. In guinea-pig myocytes dialysed with 100 microM cAMP, 74.6 +/- 3.6% of the control Cai transient was elicited by depolarisation in the presence of 5 mM Ni. The degree to which VACR triggered SR release was also graded with the pipette [cAMP] between zero and 100 microM. It therefore appears that each of the three species might possess a VACR mechanism which can be modulated by the internal [cAMP]. This may reflect an effect of cAMP to phosphorylate key proteins involved in excitation-contraction coupling. Under normal physiological conditions with a basal [cAMP] between 2 and 20 microM, VACR may play a role in triggering SR release. The role of VACR may increase under conditions which increase internal [cAMP].

Animals↗

Effect of epidermal growth factor on cultured rat hepatocytes poisoned by CCl4.

AIM: To study the effects of epidermal growth factor (EGF) on CCl4-induced primary cultured hepatocytes injury. METHODS: Alanine amino-transferase (AlaAT) and aspartate aminotransferase (AspAT) activities and K+ concentractions were determined by the Auto-biochemistry Assay System. Malondialdehyde (MDA) was determined by thiobarbituric acid method. Radioactivity was determined by liquid scintillometry. Light microscopy and electron microscopy were used. RESULTS: EGF 40 micrograms.L-1 decreased CCl4 (10 mmol.L-1)-induced damages of rat primary cultured hepatocytes by decreasing AlaAT and AspAT leakage and MDA production, and promoted RNA and DNA synthesis, with a high positive correlation between intracellular K+ leakage and DNA syntheses (r = 0.99, P < 0.01). Cytopathological study showed that EGF decreased damage of liver cells. CONCLUSION: EGF maintains the stability of cellular lipid membrane and promotes syntheses of RNA and DNA of hepatocytes, and intracellular K+ transference is a promotor of the message transmission of DNA synthesis.

Alanine Transaminase↗

Effects of daurisoline on intracellular Ca2+ activity in myocardium.

AIM: To explain the effect of daurisoline (DS) on delayed afterdepolarization (DAD). METHODS: Ca(2+)-sensitive microelectrode technic was used to record intracellular Ca2+ activity (alpha Cai) and triggered activity (TA) arising from DAD in myocardium. RESULTS: Strophantin G 3 mumol.L-1 yielded an increase in resting myocardial alpha Cai by 0.19 +/- 0.11 mumol.L-1 and transient elevations of alpha Cai by 1.48 +/- 0.55 and 4.96 +/- 1.81 mumol.L-1, respectively during the development of DAD and TA. By pretreatment with DS or verapamil, strophantin G-caused elevations of the alpha Cai in resting and provoked myocardia were eliminated and TA disappeared. DS 50 mumol.L-1 reduced Na(+)-free medium-induced elevation of dog Purkinje fibrous alpha Cai and abolished caffeine-induced increase of dog myocardial alpha Cai. CONCLUSIONS: DS inhibited DAD and TA by preventing an increase of alpha Cai via transmembrane Ca2+ entry and Ca2+ release from the reticulum.

Action Potentials↗

Exacerbation of cold restraint-induced gastric ulcer by GABA in mice.

AIM: To study the effect of GABA on cold restraint-induced gastric ulcer (CRGU) and its mechanism. METHODS: CRGU was produced in adult male mice at 1 degree C in cold room under restraint for 1 h. The ulcer index indicated the severity of gastric mucosa lesion. Gastric mucin was determined with alcian blue dye. RESULTS: GABA 2 mumol (i.c.v.) accelerated CRGU, Bicuculline (Bic) 0.2 mumol (i.c.v.) did not affect CRGU, whereas Baclofen (Bac) 4 mumol (i.c.v.) exacerbated CRGU. Bic did not modify the exacerbation of CRGU by GABA and Bac. Atropine (Atr) 0.2 mg.kg-1 (s.c.) and Phentolamine (Phen) 2.5 mg.kg-1 inhibited CRGU, and abolished the exacerbation of CRGU by GABA. Cold restraint (CR) decreased the amount of the gastric mucin, but GABA 2 mumol had no effect on CR gastric mucin. CONCLUSION: While GABA-B receptor in brain was activated, exogenous GABA exacerbated CRGU via vagal and sympathetic nerves, bearing no relation to decrease of the gastric mucin or weakening of the gastric mucosal barrier.

Adrenergic alpha-Antagonists↗

[The role of TNF-alpha in the growth and differentiation of U937 cells induced by PMA and IFN-gamma].

The effects of PMA and IFN-gamma on regulation of growth and differentiation of human monoblastic leukemic cell U937 were examined. U937 cells were stimulated by different concentrations of PMA and IFN-gamma respectively and NBT reduction assay was used to detect the differentiation of the cells. The results showed that both PMA and IFN-gamma dose-dependently induced differentiation of U937 cells into mature macrophage-like cells. The data also revealed a time-course response in the differentiation induction. Moreover, the U937 cell growth was significantly inhibited by the treatment of PMA and IFN-gamma. These results suggest that PMA and IFN-gamma coupled the regulation of U937 cell growth and differentiation. It was found that tumor necrosis factor-alpha (TNF-alpha) was expressed by the stimulated U937 cells. The specific monoclonal antibody against TNF-alpha diminished the effects of PMA and IFN-gamma on the growth and differentiation of U937 cells. Thus the endogenous TNF-alpha may involved in the mechanism of the effects of PMA and IFN-gamma on the differentiation of U937 cells. The regulatory action of the endogenous TNF-alpha on U937 cells was not due to its cytotoxic effect.

Cell Transformation, Neoplastic↗

[Antagonism of daurisoline on arrhythmogenic delayed afterdepolarization].

Standard microelectrode technique was used to study the effect of daurisoline (DS) on delayed afterdepolarization (DAD) and triggered activity (TA) in guinea pig ventricular trabeculae. DS (50 mumol.L-1) abolished TA induced by ouabain or caffeine, inhibited isoprenaline-induced TA, decreased incidence of TA induced by K(+)-free and high-Ca2+ medium or phenylephrine from 100.75% to 9.1% and 22.2% respectively, and significantly suppressed DAD caused by the above induces. It is concluded that DS exert antagonistic action to DAD and TA in guinea pig trabeculae.

Alkaloids↗

[Measurement of cytoplasmic Ca2+ activity in myocardia with Ca(2+)-sensitive microelectrode].

The improved neutral ligand ETH1001 Ca(2+)-sensitive microelectrodes (Ca-ISE, tip diameter 0.4-0.8 mumol.L-1) are highly ion selective and sensitive and therefore can be reliably used to measure cytoplasmic Ca2+ activity (alpha iCa) in myocardia. The resting alpha iCa in guinea-pig ventricular trabecula, canine ventricular myocardia and Purkinje fibers were respectively 0.19 +/- 0.01, 0.20 +/- 0.02 and 0.46 +/- 0.07 mumol.L-1. Three mumol.L-1 strophantin G increased the resting and dynamic myocardial alpha iCa by 0.18 +/- 0.02 and 6.69 +/- 2.09 mumol.L-1 respectively, as well as engendered triggered activity (TA). When pretreated with 100 mumol.L-1 dauricine (Dau), strophantin G could no longer increase alpha iCa and TA disappeared. It is suggested thus the applicability of Ca-ISE for measurement of myocardia alpha iCa and TA is obvious.

Alkaloids↗

Assessment of fermentation in growing pigs given unmolassed sugar-beet pulp: a stoichiometric approach.

In four experiments growing pigs were given a cereal-based diet alone or supplemented with unmolassed sugar-beet pulp (SBP), used as a model substrate for fermentation. The rates of production of methane and gaseous hydrogen were measured and, together with the molar proportions of volatile fatty acids (VFA) in the digesta, used in stoichiometric calculations of fermentation. The resulting estimates were only one-sixth of the observed extent of digestion of SBP. Bacteriostatic levels of antibiotics reduced fermentation by more than half, as judged from the digestion of non-starch polysaccharides: allowing for the incomplete suppression of fermentation it was estimated that the production of methane and VFA could account completely for the digested SBP. The potential contribution of various routes of hydrogen disposal to the error of the stoichiometric calculations is discussed.

Animals↗

Purification and characterization of the biologically active human truncated macrophage colony-stimulating factor expressed in Saccharomyces cerevisiae.

A human truncated macrophage colony-stimulating factor (M-CSF) cDNA encoding amino acid residues from 3 to 149 of the native M-CSF was isolated by using the polymerase chain reaction. When introduced into Saccharomyces cerevisiae by a general secretion vector pVT 102u/alpha, it directs the expression of the biologically active dimeric form of M-CSF. Through the 3 stages of purification, i.e. concentration by DEAE-cellulose column chromatography, hydrophobic chromatography on phenyl-sepharose and Mono Q fast protein liquid chromatography, the recombinant truncated M-CSF was purified as to exhibit a specific activity of 1.02 x 10(7) units/mg of protein. SDS-PAGE of this purified truncated M-CSF showed that its apparent molecular mass is 22 kDa under reducing conditions.

Amino Acid Sequence↗

Parallel stranded DNA under the scanning tunnelling microscope.

Using scanning tunnelling microscopy, we have directly observed parallel stranded DNA helixes of 43 nucleotides in length. The double helix is right-handed and has an average spacing, 17.43 A (+/- 1 S.D.: 2.30 A), and an average apparent depth, 4.79 A (+/- 1 S.D.: 1.04 A) for each groove. The average pitch of the helical turn is 34 A (+/- 1 S.D.: 3.35 A) and consists of no more than ten base pairs. The diameter of the helix is approx. 17-20 A. Our results provide direct evidence for the existence of a parallel structure of DNA in vitro and some details of its fine structure.

Base Sequence↗

Molecular dosimetry of urinary aflatoxin-DNA adducts in people living in Guangxi Autonomous Region, People's Republic of China.

Hepatocellular carcinoma is one of the five leading human cancers causing at least 250,000 deaths each year. One of the major risk factors for this disease is exposure to dietary aflatoxins, and the development of appropriate molecular dosimetry biomarkers would facilitate the identification of individuals at risk. This study was undertaken to explore the relationship between dietary intake of aflatoxins and the excretion of the major aflatoxin-DNA adduct and other metabolites into the urine of chronically exposed people. The following protocol was developed for this investigation in Guangxi Autonomous Region, People's Republic of China, where the diets of 30 males and 12 females (ages, 25-64 years) were monitored for 1 week and aflatoxin intake levels determined each day. Starting on the fourth day, total urine volumes were obtained in consecutive 12-h fractions for 3 or 4 days. High performance liquid chromatography and competitive radioimmunoassay analyses were done on each of the urine samples, and the relationships between excretion of total aflatoxin metabolites, aflatoxin-N7-guanine, aflatoxin M1, aflatoxin P1, and aflatoxin B1, and aflatoxin B1 intake values were determined. The average intake of aflatoxin B1 by men was 48.4 micrograms/day, giving a total mean exposure during the study period of 276.8 micrograms. The average daily intake by women was 77.4 micrograms/day, resulting in a total average exposure during the 7-day period of 542.6 micrograms aflatoxin B1. Initial efforts to characterize aflatoxin metabolites in urine samples were with an analysis by competitive radioimmunoassay. The analysis by linear regression of the association between aflatoxin B1 intake/day and total aflatoxin metabolite excretion/day showed a correlation coefficient of only 0.26. These findings stimulated the immunoaffinity/analytical high performance liquid chromatography analysis for individual metabolites. When the data were analyzed by linear regression analysis, the aflatoxin N7-guanine excretion and aflatoxin B1 intake from the previous day showed a correlation coefficient of 0.65 and P less than 0.000001. Similar analysis for aflatoxin M1 resulted in a correlation coefficient of 0.55 and P less than 0.00001, whereas there was no positive statistical association between exposure in the diet and aflatoxin P1 excretion, despite aflatoxin P1 being quantitatively a major metabolite. Analysis of the total aflatoxin-N7-guanine excretion in the urine during the complete collection period plotted against the total aflatoxin B1 exposure in the diet for each of the individuals, smoothing the day to day variations, revealed a correlation coefficient of 0.80 and P less than 0.0000001.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

Selective inhibition of normal murine myelopoiesis "in vitro" by a Hox 2.3 antisense oligodeoxynucleotide.

Multiple homeobox genes are expressed in haematopoietic cell lineages and their expression is cell-type specific. Thus we hypothesized that certain homeobox genes may play an important role in the process of haematopoiesis. To prove that issue, normal murine bone marrow cells were stimulated with appropriate Colony Stimulating Factors in the presence of mouse homeobox gene (Hox 2.3) sense or antisense oligodeoxynucleotides and the effects on the haematopoietic colony formation were examined. Treatment of the cells to Hox 2.3 antisense oligodeoxynucleotides led to a selective inhibition of myeloid colony formation, both in size and in numbers, but without significant effect on erythroid and megakaryocytic haematopoiesis. Exposure to Hox 2.3 sense oligodeoxynucleotides (no-oligomers), had no such effect. It was further showed that inhibition of myelopoiesis by Hox 2.3 antisense oligodeoxynucleotides was dependent on the differentiation stage of target cells. These findings demonstrated that Hox 2.3 gene plays a critical role in regulating normal murine myelopoiesis.

Animals↗

Protective and anti-arrhythmic effects of dauricine and verapamil on acute myocardial infarction in anesthetized dogs.

Dauricine (Dau) 5 mg.kg-1 and verapamil (Ver) 0.15 mg.kg-1 iv followed by infusions of 0.1 and 0.01 mg.kg-1.min-1, respectively, for 30 min, depressed the elevated coronary venous blood LDH and CPK after LAD occlusion. Dau produced antagonistic effects on acute myocardial ischemia-induced ventricular ectopic activities (VE) and ventricular tachycardia (VT). The incidences of VE and VT in Ver group and ventricular fibrillation (VF) in both groups tended to descend. The results suggested that Dau and Ver produced marked protective effects on myocardial infarction and antagonized the acute ischemic arrhythmia.

Alkaloids↗