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Biomedical subjects

X Tian

Publications and source records attributed to X Tian.

At least 55 records · Page 3Linked to original sources
PubMed2001

[Fermentation conditions for production of alkaline elastase by alkaliphilic Bacillus XE22-4-1].

A extracellular alkaline elastase producing bacterial strain Bacillus XE22-4-1, with the optimum pH of 10.0 for its growth, was isolated and screened from alkali lake of Tibet. 2% glucose and 0.25% yeast extract are respectively the appropriate carbon and nitrogen source on elastase production. Soymeal can promote elastase production. The fermentation conditions in a 2 L stirred tank were investigated. The results have revealed that the dissolved oxygen is the most effective factor on elastase production. The maximum elastolytic activity reached 266 u/mL among 48 hours of fermentation by combination of enhancing aeration and changing the mixing speed.

Bacillus↗
PubMed2000

p130Cas regulates the activity of AND-34, a novel Ral, Rap1, and R-Ras guanine nucleotide exchange factor.

We previously identified a novel murine protein, AND-34, with a carboxyl-terminal domain homologous to Ras family guanine nucleotide exchange factors (GEFs), which bound to the focal adhesion docking protein p130(Cas). Work by others has implicated both the human homologue of AND-34, BCAR3, and human p130(Cas), BCAR1, in the resistance of breast cancer cells to the anti-estrogen tamoxifen. Here we report that AND-34 displays GEF activity on RalA, Rap1A, and R-Ras but not Ha-Ras GTPases in cells. In contrast to several other Ral-GEFs, the Ral GEF activity of AND-34 is not augmented by constitutively active Ha-Ras(Val-12), consistent with the absence of a detectable Ras-binding domain. Efficient binding to AND-34 required both the Src-binding domain and a flanking carboxyl-terminal region of p130(Cas). The p130(Cas)-binding site mapped to a carboxyl-terminal sequence within the AND-34 GEF domain. Overexpression of p130(Cas), but not an AND-34-binding mutant of p130(Cas), inhibited the Ral GEF activity of co-transfected AND-34. This work identifies a new potential function for p130(Cas) and a new regulatory pathway involved in the control of Ral, Rap, and R-Ras GTPases that may participate in the progression of breast cancer cells to tamoxifen resistance.

Adaptor Proteins, Signal Transducing↗
PubMed2000

Upregulation of tyrosine hydroxylase and downregulation of choline acetyltransferase in lead-exposed PC12 cells: the role of PKC activation.

The effects of lead (Pb) on the expression of tyrosine hydroxylase (TH) and choline acetyltransferase (ChAT) were compared in relation to Pb-activation of cPKC in the PC12 cells. Exposure to 0.53 microM Pb (0.1094 ppm) increased TH but reduced ChAT activity and mRNA levels. The increase of TH activity was detectable as early as 0.5 h of exposure, reached a maximum 150% of control after 2 h, and then diminished to a steady state 135% of control between 12 and 48 h of exposure. The decrease of ChAT activity was first detectable after 2 h of Pb exposure, reached a 45% reduction after 6 h, and remained stable thereafter through 48 h of exposure. PKC activity increased 200% after 2 h and then reverted to control levels by 48 h of Pb exposure. The increase of TH activity after 2 h but not 48 h of exposure exceeded that of its mRNA. PKC inhibitor Rö32-0342 suppressed TH activity increase after 2 h of Pb exposure by 80% without affecting TH mRNA. The decrease of ChAT activity correlated with the reductions in steady-state ChAT mRNA levels at 2 and 48 h of Pb exposure and Rö32-0342 had no effect on the Pb-induced decrease of either ChAT activity or mRNA. These results demonstrate that Pb alters TH and ChAT expression in PC12 cells in a reciprocal manner, i.e., upregulates the former but downregulates the latter. PKC is not involved in Pb-induced downregulation of ChAT but does mediate the early phase of Pb-induced augmentation of TH activity, presumably through postranslational modification (phosphorylation) of the enzyme. However, this effect is short-lived due to downregulation of PKC in the course of prolonged (48 h) Pb exposure. It is concluded that, in the course of prolonged exposure, both upregulation of TH and downregulation of ChAT reflect primarily the effects of Pb at the level of gene expression through mechanisms that are not related to Pb activation of PKC.

Animals↗
PubMed2000

Developmental stage-dependent protective effect of NGF against lead cholinotoxicity in the rat septum.

The ability of nerve growth factor (NGF) to ameliorate developmental cholinotoxicity of inorganic lead (Pb) for the septal neurons was investigated by making intracerebroventricular injections of single doses of 30 microg 2.5S NGF into maternally lead-exposed suckling rats on postnatal days P2, P4, P11, or P18. Administration of NGF on P4 or later induced septal choline acetyltransferase (ChAT) activity to the same relative extent in both Pb-exposed as in control rats but failed to reverse the net reductions of ChAT activity induced by Pb. In contrast, injection of NGF at P2 completely restored ChAT activity in Pb-exposed pups to control levels by preventing the loss of ChAT-immunoreactive cells in the septum. It is concluded that although NGF retains the capacity to upregulate ChAT throughout the period of Pb exposure, it protects against the Pb-induced loss of septal cholinergic neurons only when applied within the critical period of Pb-vulnerability between postnatal days 2 and 4.

Acetylcholine↗
PubMed2000

Enantiomeric ruthenium(II) complexes binding to DNA: binding modes and enantioselectivity.

A series of enantiomerically pure polypyridyl ruthenium(II) complexes, delta- and lambda-[Ru(bpy)2 (HPIP)](PF6)2 (delta-1 and lambda-1; bpy=2,2'-bipyridine, HPIP = 2-(2-hydroxyphenyl)imidazo[4,5-f][1,10]phenanthroline), delta and lambda-[Ru(bpy)2(HNAIP)](PF6)2 (delta-2 and lambda-2; HNAIP = 2-(2-hydroxy-1-naphthyl)imidazo[4,5-f][1,10]phenanthroline), delta- and lambda-[Ru(bpy)2 (HNOIP)](PF6)2 (delta-3 and lambda-3; HNOIP = 2-(2-hydroxy-5-nitrophenyl)imidazo[4,5-f][1,10]phenanthroline), and delta- and lambda-[Ru(bpy)2(DPPZ)](PF6)2 (delta-4 and lambda-4; DPPZ= dipyridophenazine), have been synthesized. Binding behavior of these chiral complexes to calf thymus DNA (CT-DNA) has been investigated by electronic absorption, steady-state emission, and circular dichroism spectroscopies, as well as by viscosity measurements and equilibrium dialysis binding studies. Several points came from the results. (1) The DNA-binding properties were distinctly different for the [Ru(bpy)2L]2+ (L=HPIP, HNAIP, HNOIP) series of ruthenium(II) complexes, which indicates that the photophysical behavior of the complexes on binding to DNA can be modulated through ligand design. (2) Different binding rates of individual enantiomers of complexes 1 and 4 to DNA were observed through dialysis experiments. The lambda enantiomer bound more rapidly than the lambda enantiomer and their different intercalative binding geometries were suggested to be responsible. (3) Both delta-2 and lambda-2 bound weakly to CT-DNA; delta-2 may bind through a partial intercalation mode, whereas lambda-2 may bind in the DNA groove. (4) There was no noticeable enantioselectivity for complexes 1, 3, and 4 on binding to CT-DNA. Both of their enantiomers can intercalate into DNA base pairs. It is noted that delta-3 and lambda-3 exhibited almost identical spectral changes on addition of CT-DNA, and a similar binding manner of the isomers to the double helix was proposed.

Binding Sites↗
PubMed2000

A ligand-induced extracellular cleavage regulates gamma-secretase-like proteolytic activation of Notch1.

Gamma-secretase-like proteolysis at site 3 (S3), within the transmembrane domain, releases the Notch intracellular domain (NICD) and activates CSL-mediated Notch signaling. S3 processing occurs only in response to ligand binding; however, the molecular basis of this regulation is unknown. Here we demonstrate that ligand binding facilitates cleavage at a novel site (S2), within the extracellular juxtamembrane region, which serves to release ectodomain repression of NICD production. Cleavage at S2 generates a transient intermediate peptide termed NEXT (Notch extracellular truncation). NEXT accumulates when NICD production is blocked by point mutations or gamma-secretase inhibitors or by loss of presenilin 1, and inhibition of NEXT eliminates NICD production. Our data demonstrate that S2 cleavage is a ligand-regulated step in the proteolytic cascade leading to Notch activation.

Amino Acid Sequence↗
PubMed2000

Relation between left ventricular geometric alteration and extracardiac target organ damage in hypertensive patients.

To study the relation between left ventricular geometric alteration and extracardiac target organ damage in hypertensive patients. A retrospective study of 298 patients with essential hypertension was performed. Left ventricular mass index (LVMI) and relative wall thickness (RWT) were calculated using echocardiographic data. Patients were divided into four groups based on their left ventricular geometric pattern as determined using LVMI and RWT. Each of the four left ventricular geometric patterns was associated with a different degree of extracardiac organ damage. In multivariate analysis, LVMI and RWT showed strong, significant correlation to retinal changes and increases in serum creatinine levels, respectively. Alteration of left ventricular geometry resulted in an increase in the degree of extracardiac target organ damage. Echocardiographic classification of left ventricular geometry can further stratify hypertensive patients according to risk, and possibly according to the indications for intensive treatment.

Aged↗
PubMed2000

[An analysis of precision and accuracy of the phonocardiogram exercise test].

Phonocardiogram exercise testing (PCGET) is a recently developed method to evaluate cardiac contractility and the cardiac reserve of patients with heart disease and of healthy subjects. In order to test the reliability of PCGET method, the present author conducted a study on its precision and accuracy. Thirty volunteers underwent PCGET. When different examiners measured the S1 amplitude in the same cardiac cycle of the same subject, the data obtained by examiner A were: x +/- s = 5.05 +/- 0.0451; the data obtained by examiner B were: x +/- s = 4.95 +/- 0.0346, F = 1.699, P > 0.05. When different examiners measured the same cardiac cycle of the same subject, the data obtained by examiner A were: x +/- s = 0.789 +/- 0.0018; the data obtained by examiner B were: x +/- s = 0.787 +/- 0.0017, F = 1.167, P > 0.05. The results suggest that PCGET is a nonivasive, convenient, and inexpensive technique to quantitatively evaluate cardiac reserve for abnormal or normal persons.

Coronary Circulation↗
PubMed2000

Lymphocytes and perforin expression in endometrium during the menstrual cycle.

OBJECTIVE: To study the lymphocytes, perforin protein and mRNA expression in endometrium of different phases in the menstrual cycle and to explore the regularity and role of the lymphocytes and perforin in the reproductive system. METHODS: Immunohistochemical and in situ hybridization techniques were employed to demonstrate the population, phenotype and mediator (perforin) of lymphocytes in the proliferative and secretory endometrium. RESULTS: In different phases of the endometrium, there were very few CD3+ typical T lymphocytes, CD4+ helper lymphocytes and CD8+ suppressor/cytotoxic T lymphocytes, while CD56+ lymphocyte (also named NK-like cell) were abundant. CD56+ lymphocytes were absent in postmenopausal endometrium. A double immunohistochemical labelling method demonstrated the co-expression of perforin protein and CD56 antigen of cells in different phases of the endometrium. Quantitative analysis of CD56+ lymphocytes and perforin-positive cells increased in number from the proliferative to the secretory endometrium (P < 0.05). In situ hybridization analysis showed that perforin mRNA positive cells formed islands in proliferative endometrial stroma, but were scattered in secretory endometrium. There was difference between perforin gene expression and protein expression in the proliferative endometrium. CONCLUSIONS: Lymphocytes and perforin expression in the endometrium at different phases of the menstrual cycle are specific and may play an important role in the reproductive system. They may exert a positive influence on embryo implantation and be involved in endometrial stroma breakdown during menstruation.

Endometrium↗
PubMed2000

[Selection and characterization of human antibodies against vascular endothelial growth factor from human phage display antibody library].

OBJECTIVE: To select anti-VEGF (vascular endothelial growth factor) antibodies (Fab fragments) from human phage display antibody library and to identify their specification and activities. METHODS: Human immunoglobulin heavy chain and light chain genes were separately amplified by RT-PCR from human peripheral lymphocytes using family specific primers and signal sequences of immunoglobulin. Human antibody library was constructed by phage display technology, and phage Fab antibodies to VEGF were screened from this library. ELISA, Western blot and (3)H-thymidine incorporation assays were used for the specification and neutralization activities of these Fab antibodies. Sequencing analysis was carried out for further identification of the antibodies. RESULTS: The repertoire of human phage display Fab library was 1.5 x 10(8). After 4 round panning with VEGF(121), 280 clones were checked for their binding activities with ELISA and 12 clones could bind to VEGF(121) specifically. Western blot demonstrated that bacterially expressed soluble Fab could specifically recognize VEGF. Results of (3)H- thymidine incorporation showed that one clone of soluble Fab could neutralize the mitogenic activity of VEGF(165) on HUVEC. Sequencing analysis showed that the obtained V(H) gene belonged to human VH6 subgroup and the light chain was VJC rearranged human VL4 gene. CONCLUSIONS: Human anti-VEGF antibodies can be obtained from human phage display library, which provides a basis for preparation of high affinity human anti-VEGF monoclonal antibodies through antibody engineer technique.

Antibodies↗
PubMed2000

[Isolation and identification of HN-98 strain of spotted fever group rickettsiae].

OBJECTIVE: One strain of rickettsiae was isolated from a patient with unknown fever from Qiongzhong county in Hainan Province in 1998, using embryonated hen eggs and proved to be the member of rickettsiae by the methods of morphology. It was named HS-98 strain after the name of the place and the year it was isolated. METHODS: The isolate was identified by methods of Microcomplement fixation assay and OCR/RFLP and compared with known species and strains of SFGR by DNA polymerase chain reaction and DNA polymerase chainreaction and was followed by restriction endonuclease fragment length polymorphism analysis. RESULTS: Results demonstrated that HN-98 strain was antigenically and genotypically identical to R. sibirica 246. CONCLUSION: The results suggested that there existed natural foci of spotted fever in Qiongzhong Area of Hainan Province.

Adult↗
PubMed2000

[The relationship between postburn gene expression of modulators in gut associated lymph tissue and the change in IgA plasma cells].

OBJECTIVE: To investigate the relationship between the postburn change in IgA plasma cells and those of IL- 4 and IL-6 in gut associated lymph tissue (GALT). METHODS: One hundred and twenty -- five SPF mice were enrolled in the study and randomly divided into three groups -- normal control (A), microbial inoculation (B) and microbial inoculation followed by burn injury. (C) In groups Band C, the mice were gavaged with candida albicans, and the mice in group C, were inflicted with 20% TBSA III degree burn at the 14th day after inoculation. The mice were sacrificed thereafter on 1, 2 and 3 postburn days and the samples were obtained aseptically. The number of candida albicans adhering to intestinal mucosal membrane was counted. The number of IgA plasma cells in lamina propria was determined by immunohistochemical staining. The gene expression of IL-4 in Peyer patch was detected by dothybridization and that of IL-6 in lamina propria was determined by insitu hybridization. RESULTS: (1) The postburn gene expression of IL-4 in Peyer patch decreased, but the IL-6 level in intestinal lamina propria increased. (2) The number of IgA plasma cells in lamina propria decreased significantly, but the adherence of candida albicans to intestinal mucous membrane increased obviously. CONCLUSION: The decrement of postburn gene expression of IL-4 in Peyer patch might lead to IgA plasma cell decrease, resulting in the increase of candida albicans adhering to intestinal mucosal membrane.

Animals↗
PubMed2000

[Studies on chemical components of Gentiana tizuensis Franch. (I)].

OBJECTIVE: To study the chemical components of Gentiana tizuensis distributed in Qinghai Province. METHOD: Chromatography and spectral analyses were used to isolate the constituents and elucidate their structure. RESULT: Four compounds were isolated and identified as uvaol, L-(+)-l-O-methyl-myo-inositol, ursolic acid,beta-sitosterol. CONCLUSION: All these components were obtained from the plant for the first time.

Gentiana↗
PubMed1999

Glucocorticoid receptor inhibits transforming growth factor-beta signaling by directly targeting the transcriptional activation function of Smad3.

The transforming growth factor-beta (TGF-beta) family of cytokines and glucocorticoids regulate diverse biological processes through modulating the expression of target genes. Here we report that glucocorticoid receptor (GR) represses TGF-beta transcriptional activation of the type-1 plasminogen activator inhibitor (PAI-1) gene in a ligand-dependent manner. Similarly, GR represses TGF-beta activation of the TGF-beta responsive sequence containing Smad3/4-binding sites. Using mammalian two-hybrid assays, we demonstrate that GR inhibits transcriptional activation by both Smad3 and Smad4 C-terminal activation domains. Finally, we show that GR interacts with Smad3 both in vitro and in vivo. These results suggest a molecular basis for the cross-regulation between glucocorticoid and TGF-beta signaling pathways.

Animals↗
PubMed1999

Novel conformationally restricted tetracyclic analogs of delta8-tetrahydrocannabinol.

Novel analogs of (-)-delta8-tetrahydrocannabinol (delta8-THC) in which the conformation of the side chain was restricted by incorporating the first one or two carbons into a six membered ring fused with the aromatic phenolic A ring were synthesized. The affinities of the novel ligands for CB1 and CB2 indicated that the "southbound" chain conformer retained the highest affinity for both receptors.

Animals↗
PubMed1999

Analysis of differential effects of Pb2+ on protein kinase C isozymes.

Protein kinase C has been implicated as a cellular target for Pb2+ toxicity. We have previously proposed that Pb2+ modulates PKC activity by interacting with multiple sites within the enzyme. In order to further characterize the Pb-PKC interactions we compared the effects of Pb2+ on the CA-dependent and -independent protein kinase C isozymes using recombinant human PKC-alpha, PKC-epsilon, and PKC-zeta as well as the catalytic fragment of bovine brain protein kinase C, the PKC-M. The results demonstrate that, whereas at pM concentrations Pb2+ activates PKC-alpha half maximally (KAct approximately 2 pM), it has no effect on PKC-epsilon, PKC-zeta, or PKC-M activities. The activation of PKC-alpha by Pb2+ is additive with Ca2+ in a manner indicating interaction with half of the calcium activation sites. In the micromolar range of concentrations, Pb2+ inhibits all PKCs with estimated K0.5 of 1.0, 2.3, 28, and 93 microM for PKC-M, PKC-alpha, PKC-epsilon, and PKC-zeta, respectively. Examination of Pb2+ effects on PKC-M kinetics indicates a mixed type inhibition with respect to ATP and noncompetitive inhibition with respect to histone. Taken together with the results of our previous study (Tomsig and Suszkiw, J. Neurochem. 64, 2667-2673, 1995) and the evidence for the existence of two Ca2+ coordination sites Ca1 and Ca2 within the C2 domain (Shao et al., Science [Washington, D.C.] 273, 248-251, 1996), the results of the current study provide further support for a multisite Pb-PKC interaction scheme wherein lead (1) partially activates the enzyme through pM-affinity interactions with the Ca1 site and inhibits the divalent cation-dependent activity through nM-affinity interactions with Ca2 site in the C2 domain and (2) inhibits the constitutive kinase activity through microM-affinity interactions with the catalytic domain. The concentration dependence of the differential effects of Pb2+ on the calcium-dependent and -independent PKCs underscores the importance of the C2 motif as a high affinity molecular target for Pb2+.

Amino Acid Sequence↗