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Biomedical subjects

X X Zhao

Publications and source records attributed to X X Zhao.

At least 19 recordsLinked to original sources

Behaviour of Sinapis alba chromosomes in a Brassica napus background revealed by genomic in-situ hybridization.

Genomic in-situ hybridization (GISH) was applied to study the behaviour of addition chromosomes in first and second backcross (BC) progenies of hybrids between Brassica napus ssp. napus L. (AACC, 2n = 38) and Sinapis alba L. (SS, 2n = 24) produced by electrofusion. With GISH using genomic DNA of S. alba was used as probe it was possible to clearly distinguish both of the parental genomes and effectively monitor the fate of S. alba chromosomes in the BC(1) and BC(2) progenies. GISH analysis confirmed the sesquidiploid genome composition (AACCS) of the BC(1) progenies, which contained 38 chromosomes from B. napus and 12 chromosomes from S. alba. Genome painting in the pollen mother cells (PMCs) of the BC(1) plants revealed intergenomic association between B. napus and S. alba chromosomes, whereby a maximum of 4 trivalents between AC and S chromosomes were identified at metaphase I. In the BC(2) progenies, aneuploids with different numbers of additional chromosomes from S. alba, ranging from 1 to 7, were confirmed. Three putative monosomic alien addition lines were characterized, and the results are discussed with respect to the potential for intergenomic chromosome recombination.

Brassica napus↗

Preliminary observations on the concentrations of serum luteinizing hormone and follicle-stimulating hormone during gestation in bactrian camels (Camelus bactrianus).

The concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the serum of the Bactrian camel (Camelus bactrianus) throughout pregnancy were analysed by radioimmunoassay. The samples were withdrawn at intervals of half a month. The serum LH concentrations varied significantly between 2.2+/-0.9 and 20.3+/-18.8 ng/ml during the pregnancy, with the highest value on the day of artificial insemination, which indicated that a preovulatory LH peak appeared. Thereafter, the LH concentration tended to fall, reaching a fairly constant low level by the eighth month of pregnancy. The serum FSH concentrations varied between 7.0+/-0.2 and 28.9+/-0.4 ng/ml during gestation, with the peak value being reached at 4.5 months, followed by a marked drop to 7.0+/-0.2 ng/ml at 7.5 months.

Animals↗

Isolation of ovulation-inducing factors in the seminal plasma of Bactrian camel (Camelus bactrianus) by DEAE-cellulose chromatography.

Ovulation in the Bactrian camel (Camelus bactrianus) depends upon the ovulation-inducing factor in the seminal plasma; however, little research has been conducted to isolate and identify the factor. The current study attempts to isolate and identify the bioactive fractions from the seminal plasma of Bactrian camel. The seminal plasma was fractionated by diethylamino-ethylcellulose (DEAE)-cellulose chromatography and five protein fractions were obtained. The bioactive of each fraction was estimated by rat pituitary tissue culture in vitro and by the intramuscular injection of the bioactive fraction to the female camels in vivo. The concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the pituitary culture media before and 6 h after the addition of each fraction and in the peripheral blood plasma collected from the camel immediately before and hourly after the injection of the active fraction were measured by radioimmunoassay. The results demonstrated that the third fraction (L3) had the bioactive potential to stimulate the release of LH in vitro from 11.82 +/- 1.77 to 25.63 +/- 3.84 mIU/ml after the addition of L3 to the culture media. The in vivo concentrations of LH in the blood plasma of the camel increased from 6.43 +/- 0.14 before to 15.50 +/- 2.64 ng/ml 6 h after injection of L3. However, the concentrations of FSH did not show any significant changes either in vitro or in vivo. The results clearly demonstrated the existence of LH-releasing associated fractions in the seminal plasma that appears to be separated by DEAE-cellulose matrix and the isolated L3 fraction might be the ovulation-inducing factor or one of its components.

Animals↗

Reproductive aspects and storage of semen in camelidae.

The characteristics of male and female reproductive tracts and reproductive physiology in camelids are described. An account is given on methods of collection, characteristics and storage of semen, and fertility after artificial insemination (AI) with fresh, liquid-stored and frozen-thawed lamoid and camel semen.

Animals↗

Extravascular administration of interferon alfa-N3 increases serum exposure and 2-5(A) synthetase activity.

The objective of this study was to evaluate the pharmacokinetics, pharmacodynamic response, and safety of single intravenous (i.v.), intramuscular (i.m.), and subcutaneous (SQ) doses of interferon alfa-n3. Six healthy adults received 10 million units of i.v., i.m., and SQ interferon alfa-n3 in a randomized three-period crossover fashion. Serum interferon alfa-n3 concentrations and 2'-5'-oligoadenylate synthetase (2-5[A] synthetase) activity in peripheral blood mononuclear cells were determined after each dose. Extravascular administration significantly increased mean serum interferon alfa-n3 AUC values (1152 +/- 214, 944 +/- 209, and 576 +/- 188 U.h/mL, p < 0.001, with SQ, i.m., and i.v. administration, respectively) and 2-5(A) synthetase activity at 36 and 48 hours after dosing. Mild to moderate flu-like symptoms were reported by all 6 subjects, with no route-related difference in type or incidence. Interferon alfa-n3 is generally well tolerated by the i.v., i.m., and SQ routes, with i.m. and SQ administration maximizing serum exposure and 2-5(A) synthetase activity.

2',5'-Oligoadenylate Synthetase↗

A new contained human immunodeficiency virus type 1 host cell system for evaluation of antiviral activities of interferons and other agents in vitro.

HIV-host infection systems in vitro are important in the pre-clinical assessment of anti-retroviral drug activity. The present report describes the development of a new HIV-host model comprised of an epithelial cell line of HeLa lineage (HeLa-1), transfected with expression vectors bearing tat and rev (TART) genes of HIV-1 as well as the CD4 receptor gene, and HIV-1(delta Tat/Rev), a biologically contained strain of HIV-1 deleted in tat and rev. Measurement of infectivity, by syncytium formation and reverse transcriptase assay, revealed that HeLa-1 is infected with HIV-1(deltaTat/Rev). This virus failed to productively infect the TART-deficient CD4-positive HeLa cells, confirming its contained, non-infectious nature. The HeLa-1/HIV-1deltaTat/Rev system was used to measure the anti-retroviral activity of a human leukocyte-derived interferon (IFN-alphan3) preparation, several nucleoside analogs, and protease inhibitors. The HeLa-1/ HIV-1(deltaTat/Rev model provides a biologically contained system for the study of the HIV pathogenesis and the relative and combined therapeutic effects of anti-retroviral agents in vitro.

Anti-HIV Agents↗

Serum progesterone and 17 beta-estradiol concentrations during pregnancy of Bactrian camel (Camelus bactrianus).

Serum progesterone and 17 beta-estradiol during pregnancy in the Bactrian camel were measured by radioimmunoassay. Serum progesterone concentrations increased by 15 d after artificial insemination (AI) and remained elevated throughout most of gestation, the mean concentrations (3.06 +/- 0.49 to 8.51 +/- 4.80 ng/mL) were similar to those reported for many species during the same stage of pregnancy. Serum 17 beta-estradiol increased significantly from 11 m.o. of pregnancy with peak mean concentrations of 617.47 +/- 32.56 pg/mL at the 11.5 m.o.

Animals↗

Inhibitory effect of interleukin-10 on human leukocyte interferon-alpha production by Sendai virus.

Treatment of human peripheral blood leukocytes (hPBL) with Sendai virus induces significant production of human interferon-alpha (IFN-alpha). Addition of human recombinant interleukin-10 (IL-10) to hPBL in vitro prior to treatment with Sendai virus resulted in considerable inhibition of IFN-alpha production. Downregulation of IFN-alpha production was IL-10 concentration-dependent and observed at IL-10 concentrations of as low as 0.05 ng/ml, with a median effective dose (ED50) of about 5 ng/ml. Inhibition of IFN-alpha production by IL-10 occurred at an early stage of Sendai virus induction. The inhibitory effect of IL-10 on leukocyte interferon production was specific and blocked by pretreatment with neutralizing polyclonal anti-IL-10 antibody. This downregulatory effect is at the transcriptional level, since IL-10 inhibits IFN-alpha mRNA accumulation upon Sendai virus treatment. These data suggest that leukocyte IFN-alpha production is a highly regulated process that is modulated by cytokines such as IL-10 during early immunological response to infection.

Down-Regulation↗

Interferon-alpha neutralizing antibodies in HIV and chronic HCV patients treated with natural-source human leukocyte-derived interferon-alpha n3.

Human leukocyte-derived IFN-alpha n3 (Alferon N Injection) was administered subcutaneously to treat 20 patients with asymptomatic human immunodeficiency virus type 1 (HIV-1) and 141 patients with chronic hepatitis C virus (HCV) infections. The treatment of HIV-1 and HCV patients, previously untreated with any IFN preparations, did not result in development of neutralizing antibodies to IFN-alpha n3. Among 69 HCV refractory patients who were unresponsive to previous treatment with rIFN-alpha 2b, 2 had neutralizing antibodies to rIFN-alpha 2b prior to IFN-alpha n3 therapy, with no or limited cross-reactivity to IFN-alpha n3. After retreatment with IFN-alpha n3, both patients had detectable neutralizing titers to IFN-alpha n3. Additionally, 2 other patients developed low and transient neutralizing titers to IFN-alpha n3. Interferon subtype specificity of these antibodies was tested against RP-HPLC purified fractions of IFN-alpha n3, as well as rIFN-alpha 2b and rIFN-alpha 8b. Sera from patients previously treated with rIFN-alpha 2b with high antibody titers to rIFN-alpha 2b strongly reacted with the natural IFN-alpha 2b, and to a limited extent with other iFN-alpha subtypes. Neutralizing activity against IFN-alpha 2b was significantly competed out by the presence of a small amount of other interferon subtypes present in IFN-alpha n3. One patient with prior presence of antibodies to IFN-alpha 2b developed a high antibody titer to IFN-alpha 8b with limited reactivity to IFN-alpha n3. Two of the HCV refractory patients with prior neutralizing antibodies to rIFN-alpha 2b responded to IFN-alpha n3 therapy. These data suggest that the presence of neutralizing antibodies to individual IFN-alpha species will not significantly diminish the biological activity and the clinical efficacy of multi-species IFN-alpha n3.

Antibodies↗

Alpha-interferon structure and natural killer cell stimulatory activity.

Expression vectors for human alpha-interferon (Hu-IFN-alpha) J1, a site-specific mutant [Ser116]Hu-IFN-alpha J1, and Hu-IFN-alpha J/C or Hu-IFN-alpha C/J hybrids were constructed and expressed in Escherichia coli. These interferons and others were purified by immunoaffinity chromatography with a monoclonal antibody against human alpha-interferon. Their antiviral activity and ability to stimulate natural killer cell activity were determined in comparison to several other human interferons. These results provide some insight into structure-activity relationships for stimulating natural killer cells and confirm our previous conclusions that antiviral activity cannot be used to predict other activities for an individual IFN-alpha species. The observations suggest that the tertiary structure rather than any specific linear sequence of amino acids regulates the ability of the interferons to stimulate natural killer cell activity.

Amino Acid Sequence↗

High expression of tumor necrosis factor alpha cDNA in E. coli using site-specific deletion mutagenesis.

Using oligonucleotide-directed site specific mutagenesis technique, a TNF-alpha cDNA coding 157 amino acids without signal sequence and initiated with ATG was obtained. Sequencing data showed the precise deletion and insertion of the DNA fragment as designed. The modified TNF gene was expressed in E. coli with a expression vector pBV220. The expression level reached 10(8) units per liter culture. The scanning of the SDS-PAGE gel showed that the amount of the expressed TNF is about 22.8% of total soluble bacterial protein. The TNF activity can be neutralized with TNF monoclonal antibody.

Base Sequence↗

Construction and phosphorylation of a fusion protein Hu-IFN-alpha A/gamma.

A protein consisting of human (Hu)-IFN-alpha A to which the COOH-terminal 16 amino acids of Hu-IFN-gamma were fused was prepared by constructing an expression vector by oligonucleotide-directed mutagenesis. The hybrid protein Hu-IFN-alpha A/gamma was expressed under the control of phage lambda PL promoter. The protein was purified with the use of a monoclonal antibody against Hu-IFN-alpha or the COOH-terminal amino acid sequence of Hu-IFN-gamma. The purified protein exhibited a single major band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and has antiviral activity on human and bovine cells. Unlike Hu-IFN-alpha A, but similar to Hu-IFN-gamma, the hybrid Hu-IFN-alpha A/gamma can be phosphorylated by [gamma 32P]ATP and cAMP-dependent protein kinase. The phosphorylated molecule binds to the IFN-alpha/beta receptor. The introduction of a phosphorylation site into Hu-IFN-alpha A by fusion of the region of Hu-IFN-gamma which contains the phosphorylation site provides a new reagent for studies of receptor binding, pharmacokinetics, and other studies where labeled interferons are useful. Furthermore, the introduction of phosphorylation sites into proteins provides a new principle for the preparation of a wide variety of reagents for many purposes.

Amino Acid Sequence↗

[Genetic polymorphism of AcP, EsD, 6-PGD and GPT in eleven ethnic groups of China].

The genetic polymorphism of red cell acid phosphatase (AcP), Esterase D (EsD), 6-phosphogluconate dehydrogenase (6-PGD) and Glutamic pyruvic transaminase (GPT) in eleven ethnic groups of China was studied by starch gel electrophoresis. The results of 2272 tested showed that the gene frequencies of AcPB1 in Dong, Hui, Bai, Tujia, Miao, Yi, Tibetan, Man, Yao, Hani, Buyi, etc. were 0.7835, 0.7958, 0.8137, 0.7750, 0.7624, 0.8038, 0.8075, 0.8035, 0.7725, 0.6488, 0.6896, EsD1 gene frequencies were 0.6418, 0.7315, 0.6005, 0.6025, 0.6411, 0.6411, 0.6558, 0.6305, 0.6020, 0.6023, 0.6368. 6-PGDA gene frequencies 0.9279, 0.9381, 0.9387, 0.9150, 0.9356, 0.9014, 0.7764, 0.8818, 0.9851, 0.9233, 0.9410. GPT1 gene frequencies 0.4075, 0.5367, 0.5049, 0.4824, 0.5322, 0.6106, 0.6313, 0.6400, 0.3985, 0.4930, 0.3976, respectively. In addition, some rare variants were found.

Acid Phosphatase↗

Expression of a pseudogene for interferon-alpha L.

The human leukocyte interferon-alpha L (IFN-alpha L) appears to be a pseudogene because it contains a termination codon in the DNA coding for the precursor peptide (signal peptide), but the remainder of the gene codes for an interferon protein of normal length. To determine if this gene codes for an active interferon molecule, an expression plasmid was constructed for IFN-alpha L that was produced in Escherichia coli. The IFN-alpha L is active on human and bovine cells, and exhibits a trace of activity on mouse cells.

Cloning, Molecular↗