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Biomedical subjects

S Q Zhang

Publications and source records attributed to S Q Zhang.

At least 19 recordsLinked to original sources

An animal model for pregnancy-associated sleep disorder.

We studied basic sleep changes in pregnant rats in order to understand how pregnancy alters sleep. In the rat, pregnancy increased nocturnal nonREM sleep across the entire period but increased REM sleep only in the early period. By the end of pregnancy, diurnal sleep was decreased, showing that pregnancy in rats causes biphasic sleep changes as it does in humans. Termination of pregnancy returned the enhanced sleep to baseline as in the estrous cycle. Therefore, significant changes in the pattern of sleep occurred during pregnancy in rats, suggesting that the animal model may contribute to understanding the mechanism of sleep disorders related to human pregnancy.

Animals

Intracellular location of the Saccharomyces cerevisiae CDC6 gene product.

The CDC6 gene product from Saccharomyces cerevisiae is required for transition from late G1 to S phase of the cell cycle. We have investigated the subcellular localization of the CDC6 protein in yeast to explore where Cdc6p exerts its gene function (s). Using affinity-purified sera we localized Cdc6p to the cytoplasm and the nuclear matrix by both subcellular fractionation and indirect immunofluorescence microscopy. The nuclear localization was confirmed to be in the nuclear scaffold by the low-salt extraction method. The Cdc6p cannot be detected in the mitochondrial or plasma membrane fractions. Using indirect immunofluorescence, we found that a subpopulation of Cdc6p migrated into the nucleus after G1/S transition and diminished after M phase, suggesting its temporal role in nuclear DNA replication. The predicted Cdc6p polypeptide contains a conserved nuclear localization, 27PLKRKKL33, similar to that of the SV40 large T antigen and other nuclear proteins. To test whether this peptide segment plays a role in mediating nuclear transport, we have carried out site-directed mutagenesis to alter the conserved 29Lys to Thr and Arg. The wild-type nuclear localization signal of Cdc6p was found to mediate the LacZ reporter gene fused to CDC6 efficiently to the nucleus, but not the mutated versions of the nuclear localization motif. The results suggested that 29Lys is important in mediating nuclear localization, the 29Thr and 29Arg mutant versions of the CDC6 gene were also unable to complement the cdc6 temperature-sensitive mutant. However, when these mutants were expressed from a multicopy plasmid, the mutated genes could complement the mutation. Similar results were obtained in the cdc6-disrupted cells. Taken together, we suggest that (i) Cdc6p is predominantly located in the cytoplasm, (ii) the nuclear entry of Cdc6p is cell cycle dependent, and (iii) nuclear entry of Cdc6p is mediated by its nuclear localization signal. The presence of Cdc6p in both the nucleus and the cytoplasm suggests a model that Cdc6p exerts its gene function in DNA replication and mitotic restraint in the cell cycle.

Amino Acid Sequence

Hepatitis A in a Chinese urban population: the spectrum of social and behavioural risk factors.

BACKGROUND: Viral hepatitis is a major public health problem in China. Hepatitis A infections represent a substantial proportion of these, particularly in urban centres. Little is known about the social and behavioural factors in the urban household environment that influence the transmission of hepatitis A. METHODS: We conducted a register-based case-control study to investigate the risk factor patterns for hepatitis A in the general population of the City of Wuhan, in the PR China. Cases were selected from district-based health registers. One control, matched for sex and age, was identified from the case's neighbourhood. Home-based interviews combined with household observation were performed to obtain information on social, behavioural and economic risk factors and the household's indoor and outdoor environment. Analysis included conditional logistic regression. RESULTS: Hepatitis A infection was associated with a variety of social and household-related factors, like handwashing habits (after working in the garden: adjusted odds ratio [OR] = 8.24, 95% confidence interval [CI]: 1.5-44.2, before food preparation: OR = 4.68, 95% CI: 1.8-12.0; before eating: OR = 4.92, 95% CI: 1.5-15.7), and the source of fresh vegetables (OR = 3.90, 95% CI: 1.6-9.8). Hygiene in the kitchen and the household surroundings and the disposal of children's stools in vegetable gardens or refuse pits were significantly associated with univariate analysis only. The lack of possession of luxury consumer items as a surrogate indicator for income was significantly associated with the disease (OR = 2.47, 95% CI: 1.0-6.1). The study clearly established that exposure to health and hygiene education was less in the group of hepatitis A cases when compared to healthy controls (OR = 2.80, 95% CI: 0.9-8.3). CONCLUSION: The results of this study underline how social and behavioural factors are important determinants for hepatitis A in urban Chinese populations. These issues could be addressed by appropriate health and hygiene education targeted at high risk groups, and by strengthening existing procedures for monitoring and control of food hygiene.

Adolescent

Pregnancy-associated sleep changes in the rat.

Sleep disorder during the course of pregnancy has been recently recognized in humans. However, the underlying mechanism of pregnancy-associated sleep disorder remains undetermined, and sleep changes even during normal pregnancy have not been fully understood. To describe the effects of pregnancy on sleep, sleep-wake patterns before and after fertile mating were compared in an animal model. Baseline recordings of sleep and brain temperature were made throughout a normal 4-day estrous cycle in female rats. After the rats became pregnant, the recordings continued across the entire pregnant period. Compared with baseline sleep before mating, both non-rapid eye movement sleep and rapid eye movement sleep increased significantly from the first night of pregnancy. Although rapid eye movement sleep returned to the baseline level from midpregnancy, nocturnal non-rapid eye movement sleep stayed enhanced during the entire pregnant period. Daytime sleep fluctuated toward the end of pregnancy. Brain temperature was elevated during the early period of pregnancy but did not correlate with enhanced sleep. The results suggest that physiological changes in different stages of pregnancy may contribute to the regulation of maternal sleep and temperature.

Animals

Sleep patterns in cyclic and pseudopregnant rats.

In order to clarify the relationship between sleep and reproductive activities, time-course changes in sleep were analyzed in normal female rats. The state of sleep-wakefulness was continuously monitored for 4 weeks including two-consecutive 4-day estrous cycles, a 12-day pseudopregnant period and a subsequent 4-day cycle. Sleep patterns in estrous cycle were characterized by a marked reduction in nocturnal non-rapid-eye-movement sleep (NREMS) and rapid-eye-movement sleep (REMS) at proestrus. A significant increase in nocturnal NREMS and REMS occurred immediately after the induction of pseudopregnancy by sterile mating, lasting for the whole pseudopregnant period for NREMS and during the early and mid period for REMS. In contrast, diurnal REMS tended to decrease towards the end of pseudopregnancy. The circadian rhythm of brain temperature exhibited no reproductive activity-dependent change. The dynamic changes in sleep may correlate alterations in neuroendocrine activities specific to estrous cycles and pseudopregnancy.

Animals

Pulsed field gel electrophoresis labeling method to study the pattern of Saccharomyces cerevisiae chromosomal DNA synthesis during the G1/S phase of the cell cycle.

Yeast Saccharomyces cerevisiae is an excellent model to study eukaryotic DNA replication. Since yeast lacks thymidine kinase, it is difficult to assay DNA synthesis. In this report, a novel approach called the pulsed field gel electrophoresis (PFGE) labeling method is used to investigate yeast chromosomal DNA synthesis. In this method, yeast cells are first labeled by 32P in vivo and chromosomal DNA molecules are then resolved by pulsed field gel electrophoresis. A linear 32P labeling of chromosomal-size DNA molecules can be observed up to 100 min in an asynchronized culture. In an alpha-factor arresting-and-releasing synchronized culture, we observed that 32P can be rapidly taken up in the S phase. Our results show that all of the chromosomes are labeled at approximately the same time (within a 15-min interval), suggesting that the temporal order of all chromosomal DNA synthesis is synchronized in the S phase. Cell cycle blockers were used to arrest the yeast cultures to study DNA synthesis that coincides with cell cycle analysis. Our results show that alpha-factor and hydroxyurea block the cell cycle at the late G1 and S phases, respectively, resulting in the failure of DNA synthesis. Nocodazole blocks the cell cycle after the S phase and thus shows no effect on chromosomal DNA synthesis. Since the PFGE labeling method is highly specific to chromosomal DNA synthesis, we used it to examine the DNA synthesis pattern in various cell cycle mutants. All of the G1/S cdc mutants tested, cdc4, 6, 7, 8, 17, 2, and 9, cannot label the chromosomal size of DNA molecules; but the G2/M cdc mutant cdc13 can.(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutinins

Sucrose: a solute that accumulates in the guard-cell apoplast and guard-cell symplast of open stomata.

Stomatal conductances of Vicia faba leaves were recorded over a day. Coordinately, (a) guard cells dissected from leaflets were assayed, providing total sucrose content, and (b) guard cells dissected from rinsed epidermes were also assayed, providing symplastic sucrose content. Compared with that of pre-dawn samples, apoplastic sucrose content increased 4.8-5.2 x (2 experiments), reaching 1,130-1,300 fmol.guard-cell-pair-1 at midday, when conductance was highest (ca. 0.13 mol.m-2.s-1); symplastic sucrose content increased 2.5-3.5 x, reaching 350-390 fmol.guard-cell-pair-1. Thus, there is a correlation between transpiration and guard-cell sucrose content, particularly that portion localized to the apoplast. Moreover, apoplastic sucrose is apparently a source of guard-cell nutrition and, possibly, osmoticum.

Carbon Dioxide

Temporal and spatial distributions of yeast nucleoside diphosphate kinase activities and its association with the Cdc8p.

Nucleoside diphosphate kinase (E.C. 2.7.4.6.) is a broad substrate-specific enzyme that catalyzes the phosphorylation of nucleoside diphosphates to the corresponding triphosphates in nucleic acid biosynthesis. In this report, we investigate its spatial and temporal distributions in yeast to understand how the enzyme exerts its gene function(s). Our results show that the enzyme is predominantly cytoplasmic. A substantial amount of enzyme activity (40-50%) may be associated with the cell membrane. Less than 1% of total activity was detected in the nuclear fraction. Approximately 3% was found in the mitochondrial fraction. When yeast cultures were synchronized, we found that Saccharomyces cerevisiae nucleoside diphosphate kinase did not show cell cycle periodicity, as Schizosaccharomyces pombe enzyme did. To explore its link with DNA synthesis, we investigated its relationship with the Cdc8p (dTMP kinase). We demonstrated a physical interaction between these proteins in vitro, as evidenced that the GST:Cdc8p protein affinity column could retain a subpopulation of nucleoside diphosphate kinase activity from yeast crude extract. Furthermore, when GST:Cdc8p protein was expressed in yeast, the protein could bind to the glutathione-agarose, along with nucleoside diphosphate kinase, suggesting that there is an interaction between GST:Cdc8p and nucleoside diphosphate kinase in vivo. Our results provide evidence for at least a two-enzyme complex that may well facilitate nucleotide channeling in the cell.

Cell Cycle

Lessened malate inhibition of guard-cell phosphoenolpyruvate carboxylase velocity during stomatal opening.

Leaflets of Vicia faba with closed stomata or with opening stomata were freeze-dried. Excised guard-cell pairs were assayed individually under suboptimal conditions (pH 7.1 and subsaturating substrate) for phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) using quantitative histochemical procedures. L-Malate, 400 microM, significantly inhibited guard-cell PEPC activity of closed stomata but not that of opening stomata. We postulate that the lessened sensitivity of guard-cell PEPC activity to malate inhibition is an important regulatory feature of stomatal opening, which is associated with malate accumulation.

Fabaceae

Human dTMP kinase: gene expression and enzymatic activity coinciding with cell cycle progression and cell growth.

dTMP kinase (E.C.2.7.4.9.) catalyzes the phosphorylation of dTMP to the corresponding diphosphate. This enzyme is essential for DNA synthesis in vivo and is an important intermediate enzyme in the pathway of many pyrimidine analog drugs. In this report, we describe the isolation of the human dTMP kinase gene by functional complementation of a Saccharomyces cerevisiae cell cycle mutant, cdc8. The cDNA sequence revealed an open reading frame that encodes a protein with the molecular weight of 23,806. The deduced protein sequence was compared to known dTMP kinase sequences from different organisms. Although functionally complementary and structurally conserved, expressed human dTMP kinase in yeast shows little enzymatic activity. In contrast, active human dTMP kinase can be expressed from the gene cloned into the baculovirus expression system, as evidenced by increased enzymatic activity by four- to five-fold. Unlike yeast dTMP kinase, human dTMP kinase does not contain a cysteine residue after the conserved glycine-rich loop, but its enzymatic activity is still affected by the sulfhydryl inhibitor, 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB). The levels of dTMP kinase mRNA and its enzymatic activity fluctuate during the cell cycle, peaking at the S phase. Thus, like Saccharomyces cerevisiae CDC8 (encoding dTMP kinase), the human homolog mRNA and enzymatic activity are also cell cycle regulated. We have also examined four neuroblastoma cell lines for dTMP kinase mRNA levels and its kinase activities, which appear to vary according to cell growth rate. Our results suggest that the expression of the dTMP kinase gene and its activity coincide with various stages of cell growth. The identification of the human dTMP kinase gene and expression of its product in the baculovirus expression system should facilitate study of the mechanism of gene regulation and its role in pyrimidine metabolism.

Amino Acid Sequence

[Clinicopathologic and immunohistochemical study of the clear cell variant of thyroid carcinoma].

Immunohistochemical studies on 12 cases of clear cell variant of thyroid carcinoma demonstrated immunoreactivity for thyroglobulin in the clear cells of 11 tumors and for calcitonin in those of 1 tumor. This method can be helpful in differentiating primary clear cell carcinomas of the thyroid from those originating from other tissues. The clear cell variant of thyroid carcinoma exhibits a more aggressive local behavior, but satisfactory curative results can be obtained by sufficient surgical excision of the primary thyroid tumor and metastatic carcinoma of the cervical lymph nodes. The clear appearance of the tumor cells is mainly due to intracellular accumulation of large amounts of glycogen.

Adenocarcinoma, Clear Cell

Prolonged depolarization in rods in situ.

Intracellular recordings were made from rods in the superfused retina of the marine toad (Bufo marinus). It was found that injection of a brief depolarizing current pulse (0.04-1 nA) evoked a distinctive, long-lasting response, here called "the prolonged depolarization." The response appears to be regenerative, has a stereotypical waveform, is typically about 6 mV in amplitude and 3 s in duration, and has a relatively long recovery period (10-60 s). As a rule, the response cannot be directly evoked by light but the current-evoked response is significantly enhanced in the presence of steady illumination. The light-evoked hyperpolarization and the depolarizing spikes of the rod are both attenuated in the presence of the prolonged depolarization. The prolonged depolarization is not an altered manifestation of the depolarizing spikes of toad rods since both can be recorded simultaneously and steady illumination suppresses the spikes while enhancing the prolonged depolarization. The response is enhanced in chloride-free superfusate and also appears to be enhanced by the use of electrodes containing chloride. The response is markedly shortened in superfusates that lack calcium or contain 1-5 mM cobalt. On this and other evidence, it is suggested that the response may be generated by the sequential action of calcium channels and calcium-activated chloride channels. Although rarely evoked by light, the prolonged depolarization of toad rods is otherwise remarkably similar to the prolonged depolarization of turtle cones. It is proposed that the prolonged depolarization, in contrast to the feedback depolarization of cones, arises from mechanisms common to both rods and cones.

Animals

Gynandroblastoma of the ovary. A case report.

An ovarian gynandroblastoma was found in a 59-year-old woman. The tumor consisting of mixed Sertoli-Leydig cells and granulosa-theca cells demonstrated no malignant features. Bisexual cells in gynandroblastoma derived from a common gonadal mesenchyme precursor or indifferent cell. The excision of appendages with tumor or hysterectomy was sufficient.

Female

Threshold of transmission of Brugia malayi by Anopheles sinensis.

In an area of central China where Brugia malayi is transmitted by Anopheles sinensis, three villages were followed for 4 years without any control measures. Diethylcarbamazine (DEC) had been used in a control programme reducing the parasite rate in Shuiwa from 12.72 to 0.59%, in Gubomen from 3.18 to 1.55% and in Moshi from 5.62 to 2.81% (although this fell the following year to 2.23%) up until the start of the trial. The village populations, the human biting rate, the parous and the gonotrophic cycle were comparable for all the areas, yet the microfilariae rate dropped to 0.18% in Shuiwa, 0.31% in Gubomen, but not in Moshi where it increased from 2.23 to 2.43%. This indicated that Shuiwa and Gubomen were below the threshold of transmission. Due to the small number of positive cases remaining, the density measurements showed less change, but a separate study on 44 individuals followed for 6 years revealed that those above 13 microfilariae per 60 mm3 remained unchanged or increased, while those below progressively decreased their densities. The threshold of transmission of B. malayi by An. sinensis was considered to be between 1.55 and 2.23% of the population, providing no individual had a higher density than 12 microfilariae per 60 mm3.

Animals

[Tongue temperature of healthy persons and patients with yin deficiency by using thermal video].

The normal tongue originated from medical workers in our hospital and the Yin deficiency tongue originated from patients who possessed typical phenomena of Yin deficiency including smothery fever, dry mouth and so on. Determination of different area's temperature on tongue surface showed that the immediate temperature on areas of the tip, edge and medium of tongues and the delay temperature on areas of the tip, edge of tongues in patients with Yin deficiency was higher than that in healthy persons and possessed statistical significance. The tongue color in patients with Yin deficiency, the higher the tongue temperature obtained. This suggested that the degree of Yin deficiency relates to the tongue temperature. Lingual diagnosis is an essential component of treatment according to differential diagnosis in TCM. The authors consider that the determination of the tongue temperature by using thermal video will be helpful to raise the accuracy of TCM diagnosis and to appraise the real effect of Chinese herbs on patients with Yin deficiency.

Adult

[Study on biosynthesis of indigo involving transferring naphthalene plasmid DNA from Pseudomonas to E. coli].

Indigo is one of the brilliant blue dyes, which was used to be extracted from plants, but now synthesized chemically. Indigo is also produced by bacteria. In recent years, Ensley, B. D. (1983) and Mermod, N. (1986) reported a pathway producing indigo of bacteria. We are currently studying indigo formation by bacteria. Using Pseudomonas sp. S13 harboring naphthalene degradation plasmid as donor and E. coli as recipient, conjugates and transformants with the plasmid were obtained. The conjugates and transformants were not only able to grow in medium with naphthalene as the sole carbon and energy source, but also to synthesize indigo in Lennox medium. The conjugates and transformants were grown in Lennox medium at 30 degrees C for 48 hr then resulted in synthesis of indigo. The production of indigo is increased in the presence of tryptophan or indole. Indigo formation was enhanced if the bacteria was grown in a medium supplemented with either 0.1% of naphthalene or 1% of salicylic acid. The present work opens a field for the synthesis of dyes by microbes and it might shed light on the potential use in controlling environmental pollution by engineering bacteria.

Coloring Agents