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Biomedical subjects

Yang Liu

Publications and source records attributed to Yang Liu.

At least 127 records · Page 7Linked to original sources

Ex vivo expansion of hematopoietic stem cells derived from umbilical cord blood in rotating wall vessel.

Expansion of umbilical cord blood mononuclear cells (UCB MNCs) was carried out in a rotating wall vessel (RWV) bioreactor and tissue culture flasks (T-flasks) in serum-containing medium supplemented with relatively low doses of purified recombinant human cytokines (5.33 ng/ml IL-3, 16 ng/ml SCF, 3.33 ng/ml G-CSF, 2.13 ng/ml GM-CSF, 7.47 ng/ml FL and 7.47 ng/ml TPO) for 8 days. The cell density, pH and osmolality of the culture medium in the two culture systems were measured every 24h. Flow cytometric assay for CD34+ cells was carried out at 0, 144 and 197 h and methylcellulose colony assays were performed at 0, 72, 144 and 197 h. The pH and osmolality of the medium in the two culture systems were maintained in the proper ranges for hematopoietic stem cells (HSCs) and progenitors culture. The RWV bioreactor, combined with a cell-dilution feeding protocol, was efficient to expand UCB MNCs. At the end of 200 h culture, the total cell number was multiplied by 435.5+/-87.6 times, and CD34+ cells 32.7+/-15.6 times, and colony-forming units of granulocyte-macrophage (CFU-GM) 21.7+/-4.9 times. While in T-flasks, however, total cells density changed mildly, CD34+ cells and CFU-GM decreased in number. It is demonstrated that the RWV bioreactor can provide a better environment for UCB MNCs expansion, enhance the contact between HSCs and accessory cells and make the utilization of cytokines more effective than T-flask.

Bioreactors↗

The role of reactive oxygen species in the herbicide acetochlor-induced DNA damage on Bufo raddei tadpole liver.

After exposure of Bufo raddei tadpoles to acetochlor (ACETO) for 14 days, malondialdehyde (MDA) and DNA-single strand break (DNA-SSB) in livers were analyzed. An enhanced accumulation of MDA suggests that ACETO causes oxidative stress, and the significant increase in the level of DNA-SSB indicates that ACETO induces DNA damage in a dose-dependent manner as well. On the basis of the fact that oxidative stress is caused by excessive production of reactive oxygen species (ROS), and the present results, we speculate that ACETO-induced DNA damage may be a consequence of the generation of ROS. To evaluate this hypothesis, tadpoles were treated with ROS scavenger, N-acetyl-L-cysteine (NAC) or melatonin (MEL), prior to ACETO exposure. The decrease of DNA-SSB level and the increase of total antioxidant capability (TAC) show that ACETO-caused DNA damage can be attenuated by NAC and MEL. In addition, a negative correlation was observed between the extent of DNA damage and the level of TAC in tadpole liver. In conclusion, the results suggest that ACETO-induced DNA damage is mediated by ROS.

Acetylcysteine↗

Regulating the drought-responsive element (DRE)-mediated signaling pathway by synergic functions of trans-active and trans-inactive DRE binding factors in Brassica napus.

DREB1/C-repeat binding factor (CBF) is a plant-specific family of transcription factors and plays a crucial role in freeze tolerance. In the present work, two groups of drought-responsive element binding factor (DREB)-like genes were isolated from Brassica napus, named Group I and Group II. The two groups of genes were both induced by low temperature, but the expression of Group I preceded that of Group II. The Group I DREBs could specifically bind with the DRE cis-acting element and activate the expression of downstream genes, but Group II factors were trans-inactive although they still had the ability to bind with DRE, which was confirmed by electrophoretic mobility shift assay. Fluorescence quenching assays indicated that the DRE binding ability of the two groups was similar. Co-expression of Group II could depress the trans-activation activity of Group I DREB in a concentration-dependent manner. These results strongly suggested that the trans-active Group I DREBs were expressed at the early stage of cold stress to open the DRE-mediated signaling pathway in cold stress, whereas the trans-inactive Group II DREBs were expressed at the later stage to close the signal pathway in a competitive manner. The results herein provide a new insight into the regulation mechanisms of the DRE-mediated signaling pathway in response to cold stress.

Amino Acid Sequence↗

[Arthroscopic treatment of the giant cell tumor of tendon sheath in knee joint].

OBJECTIVE: To explore the technique of diagnosis and treatment of intra-articular giant cell tumor of tendon sheath (GCTTS) in knee joint, and to evaluate its clinical results. METHODS: Twelve patients with GCTTS treated during 15 years were reviewed. Nine cases were misdiagnosed with meniscus injury (6 cases) and chronic synovitis (3 cases) before operation, only 3 cases were confirmed the diagnosis of GCTTS by MRI. Complete tumorectomy under arthroscope was performed on all 12 cases. RESULTS: The results of clinical and radiographic followed-up of 9 patients for 1 to 10 years after operation was satisfactory. Clinical symptoms disappeared, range of motion and X-ray findings were normal. There was no recurrent case found. CONCLUSIONS: Arthroscopic removal of GCTTS is an effective treatment. Tumor wide resection and debriding the channel which tumor passed through are the key point to prevent the recurrence after operation.

Adult↗

Runx1 determines nociceptive sensory neuron phenotype and is required for thermal and neuropathic pain.

In mammals, the perception of pain is initiated by the transduction of noxious stimuli through specialized ion channels and receptors expressed by nociceptive sensory neurons. The molecular mechanisms responsible for the specification of distinct sensory modality are, however, largely unknown. We show here that Runx1, a Runt domain transcription factor, is expressed in most nociceptors during embryonic development but in adult mice, becomes restricted to nociceptors marked by expression of the neurotrophin receptor Ret. In these neurons, Runx1 regulates the expression of many ion channels and receptors, including TRP class thermal receptors, Na+-gated, ATP-gated, and H+-gated channels, the opioid receptor MOR, and Mrgpr class G protein coupled receptors. Runx1 also controls the lamina-specific innervation pattern of nociceptive afferents in the spinal cord. Moreover, mice lacking Runx1 exhibit specific defects in thermal and neuropathic pain. Thus, Runx1 coordinates the phenotype of a large cohort of nociceptors, a finding with implications for pain therapy.

Analysis of Variance↗

Risk stratification of colon carcinogenesis through enhanced backscattering spectroscopy analysis of the uninvolved colonic mucosa.

INTRODUCTION: Our group has been interested in applying advances in biomedical optics to colorectal cancer risk stratification. Through a recent technological breakthrough, we have been able to harness information from enhanced backscattering spectroscopy, an optics phenomenon that allows quantitative, depth-selective analysis of the epithelial microscale/nanoscale architecture. In the present study, we investigated the ability of enhanced backscattering analysis of the preneoplastic mucosa to predict risk of colon carcinogenesis. METHODS: Enhanced backscattering analysis was done on intestinal mucosa at preneoplastic time points from two experimental models of colorectal cancer: the azoxymethane-treated rat and the multiple intestinal neoplasia (MIN) mouse. Data were analyzed using two previously validated spectral markers: spectral slope and principle components. We then did a pilot study on mucosal biopsies from 63 subjects undergoing screening colonoscopy. RESULTS: In the azoxymethane-treated rat, when compared with saline-treated controls, significant changes in the enhanced backscattering markers were observed as early as 2 weeks after azoxymethane treatment (before the development of aberrant crypt foci and adenomas). Enhanced backscattering markers continued to progress over time in a manner consonant with future neoplasia. These data were replicated in the preneoplastic MIN mouse mucosa. In humans, spectral slopes in the endoscopically normal cecum, midtransverse colon, and rectum were markedly reduced in patients harboring adenomas when compared with those who were neoplasia free. CONCLUSIONS: We show, for the first time, that enhanced backscattering analysis of an aliquot of uninvolved mucosa has the potential for predicting neoplastic risk throughout the colon in both experimental colorectal cancer models and humans.

Animals↗

Identification of polysaccharides from pericarp tissues of litchi (Litchi chinensis Sonn.) fruit in relation to their antioxidant activities.

A large number of polysaccharides are present in the pericarp tissues of harvested litchi fruits. A DEAE Sepharose fast-flow anion-exchange column and a Sephadex G-50 gel-permeation column were used to isolate and purify the major polysaccharides from litchi fruit pericarp tissues. Antioxidant activities of these major polysaccharide components were also evaluated. An aqueous extract of the polysaccharides from litchi fruit pericarp tissues was chromatographed on a DEAE anion-exchange column to yield two fractions. The largest amount of the polysaccharide fraction was subjected to further purification by gel filtration on Sephadex G-50. The purified product was a neutral polysaccharide, with a molecular weight of 14 kDa, comprised mainly of 65.6% mannose, 33.0% galactose and 1.4% arabinose. Analysis by Smith degradation indicated that there were 8.7% of (1-->2)-glycosidic linkages, 83.3% of (1-->3)-glycosidic linkages and 8.0% of (1-->6)-glycosidic linkages in the polysaccharide. Furthermore, different polysaccharide fractions extracted and purified from litchi fruit pericarp tissues exhibited strong antioxidant activities. Among these fractions, the purified polysaccharide had the highest antioxidant activity and should be explored as a novel potential antioxidant.

Antioxidants↗

The conserved Ala37 in the ERF/AP2 domain is essential for binding with the DRE element and the GCC box.

Four AP2/EREBP genes encoding putative ethylene-responsive element binding factor (ERF)/AP2 domains were cloned from Brassica napus, and these genes could be induced by low temperature, ethylene, drought, high salinity, abscisic acid and jasmonate treatments. These four genes, named BnDREBIII-1 to BnDREBIII-4, were highly homologous and the 37th amino acid was the only difference among their ERF/AP2 domains. BnDREBIII-1 was demonstrated to be able to bind to both dehydration-responsive element and the GCC box and transactivate the expression of downstream genes, while BnDREBIII-4 could bind neither. Further results suggested that Ala37 might play a crucial role in the DNA binding or the stability of the ERF/AP2 domain.

Abscisic Acid↗

Production and partial characterization of bacteriocin-like pepitdes by Bacillus licheniformis ZJU12.

Bacillus licheniformis ZJU12, which was isolated from soil, could produce bacteriocin-like peptides that exhibited a broad spectrum of antagonistic activity against various species of Gram-positive bacteria and fungal pathogens, but not against Gram-negative bacteria tested except Xanthomonas oryzae pv. oryzae, a rice pathogen. The bacteriocin-like peptides were sensitive to proteinase K and trypsin. The activity was stable during temperature exposure up to 100 degrees C for 30 min, but lost completely at 121 degrees C for 15 min. The cell-free supernatant of B. licheniformis ZJU12 was shown to retain the activity within the pH range of 2-9, and the optimum pH for the activity was about 6.5. No adverse effect of the antagonistic compound to mice was observed in acute toxicity tests with the dose of 0.8 mg/20 g.

Animals↗

De novo induction of a cancer/testis antigen by 5-aza-2'-deoxycytidine augments adoptive immunotherapy in a murine tumor model.

Recent studies suggest that immunotherapy targeting specific tumor-associated antigens (TAAs) may be beneficial in cancer patients. However, most of these TAAs are tumor type specific and heterogeneous among patients, thus limiting their applications. Here, we describe the de novo induction of a cancer/testis antigen (CTA) for immunotherapy of tumors of various histologies. The murine CTA P1A, normally expressed only in a few tumor lines, could be induced de novo in all P1A-negative cancer lines of eight histologic origins in vitro and in various murine xenografts by systemic administration of 5-aza-2'-deoxycytidine. The induction of P1A expression correlated strongly with demethylation of the CpG island in the promoter region of this gene. The induced antigen was processed and presented properly for recognition by H-2L(d)-restricted P1A-specific CTLs. The combination of a demethylating agent and adoptive transfer of P1A-specific CTL effectively treated lung metastases in syngeneic mice challenged with P1A-negative 4T1 mammary carcinoma cells. These data show a novel strategy of combined chemoimmunotherapy of cancer targeting a CTA induced de novo in a broad range of tumor histologies, and support further evaluation of chromatin-remodeling agents for human cancer therapy.

Animals↗

[Application of hydroxyapatite in micro-otosurgery: experiment with guinea pigs and clinical application].

OBJECTIVE: To investigate the feasibility of hydroxyapatite (HA) middle ear prostheses in otosurgery. METHODS: In the animal experiment HA prostheses were put in the bilateral bullae of the ears of 12 healthy white guinea pigs (totally 24 ears). Three guinea pigs were killed every month 1-4 months later. Morphological examination and electron microscopy were conducted to observe grafts. In the clinical experiment HA prostheses was used in ossiculoplasty in 31 cases, reconstruction of posterior wall of external auditory canal in 5 cases, and mastoid obliteration in 11 cases. RESULTS: The HA prostheses taken out from 23 ears of the guinea pigs were with normal and complete structure, and clear margin in gross anatomy and microscopically. Only one ear suffered infection. Follow-up or the patients for 1-4 years showed excellent healing of the operated parts and an improvement of hearing threshold by about 16 dBHL. CONCLUSION: Hydroxyapatite with excellent physical and chemical properties and biological tolerance HA is a good material in ossiculoplasty, canaloplasty, mastoid obliteration, and other otological operations.

Adult↗

Determination of piceid in rat plasma and tissues by high-performance liquid chromatographic method with UV detection.

A rapid, sensitive and selective HPLC method was developed and validated for determination of piceid in rat plasma and tissues. The drug was isolated from plasma and tissues by a simple protein precipitation procedure. Chromatographic separation was performed on a C(18) column with acetonitrile-water (26:74, v/v) as mobile phase. The method was successfully applied to the pharmacokinetics and tissue distribution research after oral administration of a 50 mg/kg dose of piceid to healthy male Wistar rats. The pharmacokinetic parameters showed that piceid was quickly absorbed, distributed and eliminated within 4 h after oral administration. The tissue distribution results showed that, at 10 min, the concentrations of piceid in most tissues reached peak level except in heart and testis. The highest level of piceid was found in stomach, then in small intestine, spleen, lung, brain, testis, liver, kidney and heart. The amount of piceid in testis and heart reached the peak level at 30 min. At 120 min, the amount of piceid in all tissues decreased to a low percentage of the initial concentration. Piceid was absorbed throughout the gastrointestinal tract with considerable absorption taking place in the stomach and small intestine. There was no long-term accumulation of piceid in rat tissues.

Animals↗

Expression of qBrn-1, a new member of the POU gene family, in the early developing nervous system and embryonic kidney.

It has been shown that POU domain genes play critical roles in the development of the nervous system. We have obtained a new member of the class III POU domain genes, qBrn-1, from the cDNA library of embryonic day 5 quail and have made an extensive expression pattern analysis of qBrn-1 and qBrn-2 throughout the early embryonic development by in situ hybridization. With a specific antibody we prepared, further analysis by immunohistochemistry showed that the location of qBrn-1 protein was consistent with that of the transcripts in the early developing quail. Our results showed that both qBrn-1 and qBrn-2 were preferentially expressed in the developing central nervous system, and their transcripts were initially detected in the neural plate and later in the distinct regions of the neural tube with a stage-dependent pattern. Moreover, their expression was also detected in both notochord and neural crests. However, qBrn-1 signal, different from qBrn-2, was more widely found in the auditory pits, branchial arches, and in the mesodermal components of the developing kidney. And the expression of qBrn-1 in nephric region was earlier and wider than that of mouse Brn-1, suggesting the characteristic function of qBrn-1 in the kidney formation. The distinct dynamic expression patterns of qBrn-1 and qBrn-2 indicate multiple roles of the class III POU genes in quail neurogenesis and organogenesis.

Amino Acid Sequence↗

Epm2a suppresses tumor growth in an immunocompromised host by inhibiting Wnt signaling.

The genetic mechanisms responsible for increased incidence of lymphoma in immunocompromised individuals have not been fully elucidated. We show that, in a line of TCR transgenic TG-B mice, an insertional mutation in one allele of the Epm2a locus and epigenetic silencing of another led to a high rate of lymphoma with early onset. Overexpressing Epm2a suppressed the growth of established tumor cells and the development of lymphoma in the TG-B mice, while specific silencing of the locus increased tumorigenesis in the immune-deficient host. Downregulation of Epm2a expression is widespread among mouse and human lymphoma cell lines. Epm2a-encoded laforin is a phosphatase for GSK-3beta and an important repressor in the Wnt signaling pathway. Inactivation of Epm2a resulted in increased Wnt signaling and tumorigenesis.

Animals↗

Effects of antiglaucoma drugs on collagen gel contraction mediated by human corneal fibroblasts.

PURPOSE: Measurement of intraocular pressure is affected by the shape and thickness of the cornea, and corneal shape is thought to be maintained by contraction of corneal fibroblasts. The effects of the antiglaucoma drugs latanoprost, timolol maleate, and pilocarpine on the contraction of corneal fibroblasts cultured in a 3-dimensional collagen gel were investigated. The effects of these drugs on collagen degradation by corneal fibroblasts and their possible cytotoxicity were also examined. MATERIALS AND METHODS: Human corneal fibroblasts were cultured in a 3-dimensional gel of type I collagen and in the presence of various concentrations of latanoprost, timolol maleate, or pilocarpine for various times. Collagen gel contraction was evaluated by daily measurement of gel diameter. The extent of collagen degradation was determined by measurement of the amount of hydroxyproline generated by acid-heat hydrolysis of culture supernatants. The release of lactate dehydrogenase from corneal fibroblasts was determined as an index of drug cytotoxicity. RESULTS: Latanoprost stimulated collagen gel contraction mediated by corneal fibroblasts in a concentration- and time-dependent manner, whereas timolol maleate and pilocarpine had no such effect. None of the 3 drugs affected collagen degradation by corneal fibroblasts or exhibited cytotoxicity at concentrations as high as 100 muM. CONCLUSIONS: Among the antiglaucoma drugs examined, only latanoprost stimulated collagen gel contraction mediated by human corneal fibroblasts. This action of latanoprost might affect corneal shape and thereby influence measurement of intraocular pressure.

Aged↗

Azospirillum melinis sp. nov., a group of diazotrophs isolated from tropical molasses grass.

Fifteen bacterial strains isolated from molasses grass (Melinis minutiflora Beauv.) were identified as nitrogen-fixers by using the acetylene-reduction assay and PCR amplification of nifH gene fragments. These strains were classified as a unique group by insertion sequence-PCR fingerprinting, SDS-PAGE protein patterns, DNA-DNA hybridization, 16S rRNA gene sequencing and morphological characterization. Phylogenetic analysis of the 16S rRNA gene indicated that these diazotrophic strains belonged to the genus Azospirillum and were closely related to Azospirillum lipoferum (with 97.5 % similarity). In all the analyses, including in addition phenotypic characterization using Biolog MicroPlates and comparison of cellular fatty acids, this novel group was found to be different from the most closely related species, Azospirillum lipoferum. Based on these data, a novel species, Azospirillum melinis sp. nov., is proposed for these endophytic diazotrophs of M. minutiflora, with TMCY 0552(T) (=CCBAU 5106001(T) = LMG 23364(T) = CGMCC 1.5340(T)) as the type strain.

Acetylene↗

Expression of serine threonine kinase 15 is associated with poor differentiation in lung squamous cell carcinoma and adenocarcinoma.

Serine threonine kinase 15 (STK15, also named BTAK, Aurora-A, aurora-2, or AIKI) is a type of mitotic kinase. The overexpression of STK15 is significantly associated with carcinogenesis in many tumors. The purpose of the present study was to investigate the expression of STK15 in lung squamous cell carcinoma and adenocarcinoma and analyze the correlation between STK15 expression and clinicopathological factors. The expression patterns of STK15 were examined by immunohistochemistry in 80 lung squamous cell carcinomas and adenocarcinomas and 20 normal lung tissues. The protein and mRNA expression of STK15 were evaluated by western blot and reverse transcription-polymerase chain reaction (RT-PCR) in 40 lung cancer samples and corresponding normal lung tissues. Immunohistochemically, the positivity of STK15 expression was 68.75% (55/80). The STK15 expression was significantly higher in poorly differentiated lung cancers than in well-differentiated or moderately differentiated lung cancers (P = 0.011). Western blot and RT-PCR showed that the protein and mRNA expression of STK15 were correlated (P = 0.044) and significantly higher in tumors than in corresponding normal lung tissues (P < 0.05). These results indicate that the overexpression of STK15 contributes to the carcinogenesis and de-differentiation of lung cancers.

Adenocarcinoma↗

Identification and characterization of glnA promoter and its corresponding trans-regulatory protein GlnR in the rifamycin SV producing actinomycete, Amycolatopsis mediterranei U32.

The genetic requirements for the transcription of glnA, encoding the major glutamine synthetase in a rifamycin SV-producing Amycolatopsis mediterranei strain, U32, were investigated. Primer extension experiments showed that the promoter of U32 glnA (pglnA) was likely to have two transcription initiation sites: P(1) and P(2), located 157 and 45 nucleotides (nt) upstream of the translational start codon, respectively. Gel mobility shift and DNase I footprinting analyses revealed a 30 bp cis-element located at 45 to 75 nt downstream of P1, or 38 to 68 nt upstream of P(2). The sequence of the cis-element displayed high similarity to the corresponding regions of pglnA from Streptomyces coelicolor and S. roseosporus. With xylE as a reporter gene, the expression levels of U32 pglnA and its deletion derivatives under different nitrogen-source conditions were analyzed by detecting the catechol dioxygenase activities in S. lividans TK54, S. coelicolor J508 and S. coelicolor FS10 (glnR mutant). These in vivo studies showed that the activation of U32 pglnA in S. coelicolor required GlnR, and its binding to the U32 pglnA was further confirmed by the gel mobility shift assay. Cloning and heterologous expression of the U32 glnR allowed us to detect the in vitro interaction between the U32 GlnR and the corresponding pglnA cis-element. Further evidence shown by in vivo glnR inactivation and complementation indicated that GlnR is essential for the active transcription of glnA in U32.

Actinobacteria↗