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Biomedical subjects

Z Liang

Publications and source records attributed to Z Liang.

At least 19 recordsLinked to original sources
PubMed1992

Targeting of Sp1 to a non-Sp1 site in the human neurofilament (H) promoter via an intermediary DNA-binding protein.

The human neurofilament (H) promoter contains multiple binding sites for nuclear proteins including a Proximal (Prox) site centered around the sequence GGTTGGACC and an adjacent pyrimidine (Pyr) tract site centered around the sequence CCCTCCTCCCC. Surprisingly binding to a probe containing the Prox/Pyr region of the NF(H) promoter was competed in gel shifts by an oligonucleotide containing only an Sp1 binding site (GGGGCGGGG). Supershift assays with a polyclonal anti-Sp1 antisera confirmed that Sp1 was part of the complex formed with the Prox/Pyr probe. However neither bacterially expressed Sp1 516C or vaccinia virus expressed full-length Sp1 778C bound to the Prox or Pyr sequences in DNase I footprints or gel shift assays. Gel shift competitions and supershift assays with probes containing either Prox or Pyr tract sites alone demonstrated targeting of Sp1 to the Prox binding site and identified a non-Sp1 containing complex which contains a Prox binding protein. Adding exogenous Sp1 to a HeLa nuclear extract enhanced the Sp1-containing complex but had no effect on the Prox complex. These studies show that Sp1 can be targeted to a non-Sp1 site in the human NF(H) promoter through protein/protein interactions with a distinct sequence specific DNA-binding protein.

Base Sequence↗
PubMed1992

Proton transfer roles of lysine 64 and glutamic acid 64 replacing histidine 64 in the active site of human carbonic anhydrase II.

The CO2 hydration activities of cloned human carbonic anhydrase II (carbonate hydro-lyase, EC 4.2.1.1) and variants with Lys, Glu, Gln or Ala replacing His at sequence position 64 have been measured in a variety of different buffers in the pH range 6-9. The variants with Lys-64, Gln-64 and Ala-64 showed non-Michaelis-Menten behavior under some conditions, apparent substrate inhibition being prominent near pH 9. However, asymptotic Michaelis-Menten parameters could be estimated for the limit of low substrate concentrations. All variants show distinct buffer specificities, and imidazole derivatives, Ches and phosphate buffers yield higher kcat values that Bicine, Taps and Mops buffers under otherwise similar conditions. These results are interpreted in terms of different pathways for a rate-limiting proton transfer. In unmodified enzyme, the very high catalytic activity depends on His-64 functioning as an efficient proton transfer group, but this pathway is not available in the variants with Gln-64 and Ala-64. Imidazoles, Ches and phosphate are thought to participate in a metal center-to-buffer proton transfer pathway, whereas Bicine, Taps, Mops and Mes appear to lack this capacity, so that the rate-limiting proton transfer occurs in a metal center-to-bulk water pathway for these variants. The Lys-64 and Glu-64 variants give significantly higher kcat values in Taps, Mops and Mes buffers than the Ala-64 and Gln-64 variants. The pH dependencies of these kcat values are compatible with the hypothesis that Lys-64 and Glu-64 can function as proton transfer groups. Thus, at pH near 9, Lys-64 appears to be only 5-times less efficient than His-64, while Glu-64 is inefficient. At pH 6, Lys-64 is an inefficient proton transfer group, but Glu-64 is only 2-3-times less efficient than His-64. The data indicate that Lys-64 and Glu-64 have pKa values near 8 and below 6, respectively.

Binding Sites↗
PubMed1992

Background correction by wavelength modulation for pulsed-laser-excited atomic fluorescence spectrometry.

Instrumentation was constructed to modulate the dye laser wavelength for background correction in laser-excited atomic fluorescence spectrometry (LEAFS). To achieve wavelength modulation a piezoelectric pusher was used to drive the wavelength tuning mirror in a laboratory-constructed grazing incidence dye laser. The laser pulses were synchronized with the piezoelectric pusher movement so that alternate laser pulses measured the atomic fluorescence signal at the analytical atomic spectral line (on-line) and the background signal at a wavelength displaced to one side of the atomic line (off-line). The background-corrected signal was obtained by subtracting the off-line "background" from the on-line "signal plus background". The spectral line width (fwhm) of the dye laser was 0.003 nm, while the wavelength modulation interval was controllable over the range from 0 to 0.2 nm with a spectral resolution limited only by the spectral line width of the laser. This type of background correction could, in principle, be applied to other types of tunable lasers such as pulsed Ti: sapphire lasers. The performance of background correction by wavelength modulation (WM) was demonstrated by measurement of sodium resonance fluorescence in an air-acetylene flame and by thallium nonresonance fluorescence in a graphite furnace. The experimental data indicated that the wavelength modulation corrected, effectively and quantitatively, for flame background, blackbody emission from a graphite furnace, and scatter of laser radiation off aluminum chloride (1 mg/mL as AI) matrix particles in both the furnace and the flame. Analytical results were in good agreement with certified values for the determination of sodium in standard reference materials by the use of modulated LEAFS.

Lasers↗
PubMed1992

Expression of human mid-sized neurofilament subunit in transgenic mice.

We have created transgenic mice which carry and express the gene encoding the human NF(M) subunit. RNAase protection assays reveal that the transgene is abundantly expressed in CNS and PNS but also, at very low levels in some non-neural tissues as well. Although the neurospecificity of transgene transcription was not absolute, we are able to detect the protein only in neurons with immunocytochemical techniques. Glial and endothelial cells do not contain immunoreactive materials. Interesting subtle differences in the relative level of the human transgene encoded and endogenous murine encoded NF(M) proteins were noted in different regions of the brain. Similar differences were found in the levels of transgene and endogenous gene mRNA suggesting that these differences may be traceable to differences in RNA transcription or stability. Our data demonstrate, within the sensitivity of the immunocytochemical techniques we used, that the human NF(M) protein is present only in the neurons of the transgenic mice and that it is present in the same neurons as the endogenous NF(M). Furthermore, immunoelectron-microscopic examination of isolated neurofilaments shows that the human NF(M) coassembles with the endogenous NF(M) during filament formation. Thus, although the human NF(M) possesses a much larger multiphosphorylation site in its carboxy terminus, it seems to be the functionally equivalent to the mouse protein, even in the murine neuron.

Animals↗
PubMed1992

Novel DNA binding proteins participate in the regulation of human neurofilament H gene expression.

By a combination of DNase I footprinting, methylation interference, and gel shift analyses we have identified multiple binding sites for nuclear proteins within the promoter region of the human neurofilament H gene. Two sites likely bind the transcription factor Sp1 while two others may be targets for previously unrecognized DNA binding proteins. One site, PAL, occurs within the 10 bp sequence GGGGAGGAGG. Two copies of the PAL sequence form an interrupted palindrome around one of the Sp1 sites. A second site, PROX, is found within the sequence GGTTGGACC. Nuclear extracts prepared from both neural and non-neural cell lines, mouse brain, and mouse liver contain proteins that recognize and bind to the PROX and PAL sequences indicating that proteins which bind to these target sequences are widespread. The appearance of these target sequences in the 5' upstream region of several neuron specific genes suggests that they play key roles in the transcription of neuron specific genes. The functional activity of these target DNA sequences was demonstrated by transfection assays using a reporter gene fused to nested deletions of the NF(H) promoter region. Interestingly, these assays revealed that maximal transient expression was obtained with DNA fusion genes containing the PAL, PROX and TATA sequences. Inclusion of the Sp1 sites into the fusion genes failed to enhance the expression of the reporter gene. To determine if the NF(H) promoter can be activated in a tissue specific manner during development transgenic mice containing the promoter region linked to a beta-galactosidase reporter gene were generated. In one line sporadic expression of the transgene occurred in the CNS and testis while in four other lines no expression occurred. Collectively these results suggest that the NF(H) gene promoter is active in a tissue specific manner only by interactions with regulatory elements that lie further upstream or downstream of the start site of initiation.

Animals↗
PubMed1992

Multiple nuclear factors interact with the promoter of the human neurofilament M gene.

In order to identify potential regulatory elements of the human mid-sized (M) neurofilament (NF) gene we preformed DNase I footprinting, gel mobility shift assays and methylation interference studies with probes from the NF(M) immediate 5' flanking region. These studies identified multiple sites for DNA-binding proteins including four Sp1 sites, and single sites each for members of the NF-1 and AP-1 families of DNA binding proteins. In addition a binding site within a pyrimidine tract likely binds a novel DNA-binding protein which also interacts with the human NF(H) gene promoter. Factors that bind to these sites are found in both neural and non-neural cells suggesting that the NF(M) promoter may not contain tissue specific regulatory signals. In transient assays, addition of these binding sites to an NF(M) minimal promoter containing only a TATA box lead to a greater than 40-fold activation of transcription over background. Progressive 5' deletions reduced expression in a step wise manner suggesting that all the factors likely act synergistically as positive regulators of transcription.

Base Sequence↗
PubMed1992

Simultaneous compensation for attenuation, scatter and detector response for SPECT reconstruction in three dimensions.

A three-dimensional reconstruction method for simultaneous compensation of attenuation, scatter and distance-dependent detector response for single photon emission computed tomography is described and tested by experimental studies. The method determines the attenuation factors recursively along each projection ray starting at the intersected source voxel closest to the detector. The method substracts the scatter energy window data from the primary energy window data for scatter compensation. The detector response is modelled to be spatially invariant at a constant distance from the detector. The method convolves source distribution with the modelled response function to compensate for the smoothed by use of a non-uniform entropy prior to searching for the maximum a posteriori probability solution. The method was tested using projections acquired from a chest phantom by a three-headed detector system with parallel hole collimators. An improvement was shown in image noise, recognition of object sizes and shapes, and quantification of concentration ratios.

Image Processing, Computer-Assisted↗
PubMed1992

[The effect of acupuncture on rabbits with fever caused by endotoxin].

Forty-eight rabbits were used to investigate the effect of puncturing GV14, LI11 on the change of temperature and the level of plasma endotoxin. The animals were divided into 5 groups: reinforcing manipulation, reducing manipulation, electroacupuncture, endotoxin-control, N.S.-control group. The result shows that there is no influence on the fever caused by endotoxin and the level of plasma endotoxin in the treatments by the different acupuncture techniques.

Acupuncture Therapy↗
PubMed1992

[The influence of different acupuncture manipulations on the plethysmogram of the patient with kidney deficiency].

In order to prove the specific property between the tonifying manipulation (TM) and reducing manipulation (RM), and search further for the key in improving the effect of manipulation, the influence of to on two different manipulations. The plethysmogram in the same body-condition (kidney-deficiency) One was TM with slow-trusting and quick lefting; the other was RM with the converse movement. Randomized block design, cross-over design and single-blind design were used. The result showed that the TM increased the amplitude of the plethysmogram obviously, whereas the RM had no significant influence on it. There was significant difference not only between the TM group and the central group, but also between the TM group and RM group (P less than 0.01 or P less than 0.05). The difference between the RM group and the control group was not significant (P greater than 0.05). Meanwhile, it was recommended that "mind concentration" on acupuncture manipulation is important. In conclusion, the experiment demonstrated the experimental evidence for explaining the mechanism of TM and RM. It was suggested that acupuncture manipulations are worthy to pay attention so as to improve clinical effect.

Acupuncture Therapy↗
PubMed1991

Laser-excited atomic fluorescence in a flame as a high-sensitivity detector for organomanganese and organotin compounds following separation by high-performance liquid chromatography.

The coupling of a high-performance liquid chromatograph with a sensitive and selective laser-excited atomic fluorescence spectrometry (LEAFS) detector is described. In connection with this, a study of the signal and noise characteristics of instrumentation for dispersive, nondispersive, and front surface LEAFS is reported together with a comparison of the sensitivity and selectivity achieved with high-performance liquid chromatography (HPLC)-flame LEAFS, HPLC-ultraviolet (UV), and HPLC-continuum source excited flame atomic fluorescence spectrometry (AFC) instrumentation. The HPLC-flame LEAFS instrumentation was applied to an investigation of the Mn species responsible for (methylcyclopentadienyl)manganese tricarbonyl (MMT) toxicity in rats. The detection limits for various organomanganese species by HPLC-flame LEAFS ranged from 8 to 22 pg of manganese. Recovery of these compounds from rat urine varied between 80% and 100%, with a reproducibility of between 4% and 8% relative standard deviation. Preliminary data for the HPLC-flame LEAFS determination of toxic alkyltin compounds are reported.

Chromatography, High Pressure Liquid↗
PubMed1991

Pulse timing and optical interface between a neodymium: yttrium aluminum garnet laser and a Fourier transform ion cyclotron resonance mass spectrometer.

Laser desorption/ionization combined with pulsed (time-of-flight or Fourier transform ion cyclotron resonance) mass spectrometric detection is a powerful technique for analysis of involatile compounds and mixtures. Such experiments were originally conducted with pulsed CO2 lasers. Although a pulsed CO2 laser can be operated in single-shot mode, Nd: YAG lasers perform best with multiple flashes for warm-up before the final Q-switch output light pulse, thus creating the need to synchronize the desired final laser-output pulse with the event sequence for mass spectrometric analysis. In this paper, we describe a new and simple interface (both optical and electronic components) between a Continuum (formerly Quantel) Model YG 660A Nd:YAG laser and an Extrel FTMS-2000 mass spectrometer. The optics are modified from a prior pulsed CO2 laser interface from Extrel. Synchronization between the Nd:YAG laser and the mass spectrometer event sequence is achieved by means of a simple timing circuit that uses an inexpensive pulsing device and is triggered by pulses generated directly from the Extrel 1280 data system and cell controller, in contrast to the only prior published method that required an auxiliary microcomputer. The present interface method is highly flexible, and makes possible complex sequence events involving laser pulses for e.g.: desorption/ionization of solids; photoionization of gaseous neutrals; and photodissociation and photodetachment of gaseous ions.

Lasers↗
PubMed1991

[Preliminary study of cervicovaginal peroxidase as indicator for ovulation].

The study was designed to determine whether the change in the pattern of cervicovaginal peroxidase activity in the fertile period of women can serve as a basis for the development of a technique to predict ovulation time in natural family planning. Eight healthy women volunteered for this study in Chengdu area. The samples of cervical mucus and vaginal secretion were collected. The specific G-Px activity was measured with reference to the procedure of Tsibris, et al., on days 6, 8, 10, 12 through 16, 18, 20, 22, 24 of each menstrual cycle; at the same time an antecubital venous blood sample was drawn to be analyzed for levels of LH, FSH, E2, and P by RIA. The data showed that in 7 ovulatory cycles the G-Px activity did not vary regularly in vaginal secretion but it did drop rapidly in cervical mucus 2 or 3 days before ovulation time and it rose again one day after ovulation. The results suggest that the measurement of the G-Px activity in cervical mucus can be used to predict ovulation by two to three days before the time.

Cervix Mucus↗
PubMed1991

[Evaluation of a logistic regression model in predicting the prognosis of Graves' disease treated by antithyroid drugs].

One hundred and twelve new cases of Graves' disease were treated by tapazole for 6 months and followed up for another 12 months. The initial dose was 30 mg/d. Clinical and biochemical euthyroidism was achieved within 1 to 3 months, then a maintenance was given until cessation of drugs at 6 months. One hundred and eleven cases completed the study. Remission and relapse were defined at the end of follow-up for 1 year according to the presence or absence of clinical manifestation of hyperthyroidism and the levels of T3 and T4. The results of the 12-month follow-up showed that 46 of 111 cases were in remission, and the remaining 65 cases suffered relapse. A logistic regression model with 4 variables was established, which included thyroid suppression rate and goitre size by palpation at the end of drug treatment, the level of T3 before therapy, and the patients' age. The model had 81.5% sensitivity, 84.8% specificity and 82.9% (92/111) accuracy in predicting the outcome of Graves' disease after withdrawal of drug for 1 year. The results were much better than any other univariate analysis in this study.

Adolescent↗
PubMed1991

[Intrauterine adhesion and ovulatory disorder following induced abortion].

Follicular stimulating hormone, luteinizing hormone, prolactin, estradiol and progesterone were measured by radioimmunoassay in 34 patients with those from 34 normal cycling women after induced abortion. The result showed that the anovulatory incidence of the patients (20.6%) was significantly higher than that of the controls (2.8%) (P less than 0.05). Increased prolactin and premature ovarian failure might be responsible for the anovulatory disorder, resulting in unsatisfactory restoration of fertility. It is suggested that the low success rate in treating patients with intrauterine adhesion after induced abortion may be due to premature ovarian failure and hyperprolactinaemia.

Abortion, Induced↗
PubMed1991

[Further study on the treatment of severe beta-thalassemia with myleran].

In a previous paper we reported the results of treatment of five patients with beta-thalassemia major with myleran. Three more cases are reported in this paper. After 40-50 days of oral administration of myleran, major hematological findings (Hb, HbF, RBC) began to increase and remained at higher level for 4-5 months before dropping back to the original. The results of analysis of sister chromatid exchange (SCE) showed no irreversible damage on chromosomal DNA of the patients at the dosage used (0.2 mg/kg.d). It was also demonstrated that the effect of treatment does not seem to be related to the types of beta-globin gene mutations of the patients.

Busulfan↗
PubMed1990

[Observation of the outcome of 112 cases with Graves' disease treated by short-term antithyroid drug therapy for one year].

One hundred and twelve new cases with Graves' disease were treated by tapazole for 6 months and followed-up for another 12 months in a prospective study. One hundred and eleven cases completed the whole course of study, and only one failed to be followed up. The results of the follow-up for 12 mon showed that the remission rate was 41.4% (46/111) and relapse rate was 58.6% (65/111). The present study indicated that the remission or relapse was related with serum levels of T3 before treatment, shrank goiter and disappearance of goiter bruit during the treatment as well as thyroid suppression rate, but not related to sex, age, period of illness before therapy or severity of the disease. It is suggested that a 6-mon antithyroid drug therapy instead of traditional long term therapy may be suitable for those patients who are with a reduced goiter and disappeared goiter bruit during the antithyroid therapy, and suppressible thyroid up-take by T3 and whose serum levels of T3 before treatment are not very high. So the patients may benefit from the short-term therapy. They are not only able to obtain a prolonged remission but also save time and money.

Adolescent↗