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Biomedical subjects

Li Tan

Publications and source records attributed to Li Tan.

At least 19 recordsLinked to original sources

High-efficiency stepwise contraction and adsorption nanolithography.

A new miniaturization protocol is demonstrated using stretching and relaxation of an elastomer substrate. A designed microstructure is formed on the stretched substrate and subsequently becomes miniaturized when the substrate relaxes. More importantly, the miniaturized structures can be transferred onto a new substrate for further miniaturization or can be utilized as stamps for nanolithography of designated materials. As an example of this approach, an elastic mold was first cast from a Si mold containing periodic line arrays of 1.5-microm line width. Upon relaxation, line width is reduced to 240 nm. The new elastomer may be used as stamps for micro- and nanofabrication of materials such as proteins. The polymer surface roughness or wrinkling behavior at nanoscale is found to follow classic stability model in solid mechanics. This observation provides means to design and control the surface roughness to meet specific requirements.

Acrylic Resins↗

Yersinia pestis YrbH is a multifunctional protein required for both 3-deoxy-D-manno-oct-2-ulosonic acid biosynthesis and biofilm formation.

Bubonic plague is transmitted by fleas whose feeding is blocked by a Yersinia pestis biofilm in the digestive tract. Y. pestis also block feeding of Caenorhabditis elegans by forming a biofilm on the nematode head, making the nematode an experimentally tractable surrogate for fleas to study plague transmission. Arabinose 5-phosphate isomerase (API), encoded by Y. pestis yrbH, catalyses the conversion of ribulose 5-phosphate into arabinose 5-phosphate (A5P), the first committed step in the 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) biosynthesis pathway. Here we show that Y. pestis YrbH is a multifunctional protein required for both Kdo biosynthesis and biofilm formation on C. elegans. The YrbH protein contains four functional components: biofilm-related region 1 (B1), a sugar isomerase domain (SIS), biofilm-related region 2 (B2) and a cystathionine beta-synthase domain pair (CBS). B1, SIS and B2 are all required for API function, but any of the three is sufficient for a biofilm-related function. The CBS domain appears to negatively regulate the biofilm-related function.

Aldose-Ketose Isomerases↗

The effect of glutamine-supplemented total parenteral nutrition on nutrition and intestinal absorptive function in a rat model.

The aim of the present study was to evaluate the effect of short-term (7 days) glycyl-glutamine-supplemented total parenteral nutrition (TPN) on nutrition and intestinal absorptive function in a rat model. Thirty Wistar rats, weighting 140-180 g, were divided into three groups (n=10) randomly. The animals received isonitrogenous and isocaloric TPN solutions for 7 days. The nitrogen was supplied by glycyl-glutamine dipeptide-supplemented amino acid solution (group G), and two standard amino acid solutions (group V, group N), respectively. Body weight, plasma glutamine level, nitrogen balance, total tissue water and intestinal absorptive function, assessed by (15-N)-glycine absorption, were investigated. Body weight decreased in three groups at the end of TPN; there was no significant difference in relative body-weight changes. There was a significant improvement of cumulative nitrogen balance and nitrogen retention in group G compared to other groups (P<0.05). There was no significant difference in intestinal glycine absorption (P>0.05) among the three groups. Total tissue water of left thigh muscle was significantly higher in group V and group N than that in group G (P<0.05). The results indicated that short-term (7 days) TPN supplemented with glycyl-glutamine improved plasma glutamine level and nitrogen balance, decreased water content of muscle, but had no beneficial effect on absorptive function in a rat model.

Analysis of Variance↗

Effect of docosahexaenoic acid on interleukin-2 receptor signaling pathway in lipid rafts.

Recent studies have shown that polyunsaturated fatty acids (PUFA) regulated the functions of membrane receptors in T cells and suppressed T cell -mediated immune responses. But the molecular mechanisms of immune regulation are not yet elucidated. Lipid rafts are plasma membrane microdomains, in which many receptors localized. The purpose of this study was to investigate the effect of DHA on IL-2R signaling pathway in lipid rafts. We isolated lipid rafts by discontinuous sucrose density gradient ultracentrifugation, and found that DHA could change the composition of lipid rafts and alter the distribution of key molecules of IL-2R signaling pathway, which transferred from lipid rafts to detergent-soluble membrane fractions. These results revealed that DHA treatment increased the proportion of polyunsaturated fatty acids especially n-3 polyunsaturated fatty acids in lipid rafts and changed the lipid environment of membrane microdomains in T cells. Compared with controls, DHA changed the localization of IL-2R, STAT5a and STAT5b in lipid rafts and suppressed the expression of JAK1, JAK3 and tyrosine phosphotyrosine in soluble membrane fractions. Summarily, this study concluded the effects of DHA on IL-2R signaling pathway in lipid rafts and explained the regulation of PUFAs in T cell-mediated immune responses.

Docosahexaenoic Acids↗

Identification and genome characterization of Heliothis armigera cypovirus types 5 and 14 and Heliothis assulta cypovirus type 14.

Genomic characterization of Heliothis armigera cypovirus (HaCPV) isolated from China showed that insects were co-infected with several cypoviruses (CPVs). One of the CPVs (HaCPV-5) could be separated from the others by changing the rearing conditions of the Heliothis armigera larvae. This finding was further confirmed by nucleotide sequencing analysis. Genomic sequences of segments S10-S7 from HaCPV-14, S10 and S7 from HaCPV-5, and S10 from Heliothis assulta CPV-14 were compared. Results from database searches showed that the nucleotide sequences and deduced amino acid sequences of the newly identified CPVs had high levels of identity with those of reported CPVs of the same type, but not with CPVs of different types. Putative amino acid sequences of HaCPV-5 S7 were similar to that of the protein from Rice ragged stunt virus (genus Oryzavirus, family Reoviridae), suggesting that CPVs and oryzaviruses are related more closely than other genera of the family Reoviridae. Conserved motifs were also identified at the ends of each RNA segment of the same virus type: type 14, 5'-AGAAUUU...CAGCU-3'; and type 5, 5'-AGUU...UUGC-3'. Our results are consistent with classification of CPV types based on the electrophoretic patterns of CPV double-stranded RNA.

Amino Acid Sequence↗

[Sensitization of human glioma SWO cell line to tumor necrosis factor-induced apoptosis by blocking phospholipase C-gamma1 signaling pathway].

OBJECTIVE: To explore the role of phospholipase C-gamma1 (PLC-gamma1) in tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis of human glioma SWO cells. METHODS: The PLC-gamma1 pathway was blocked by U73122 in SWO cells, and the inhibitory effect of TNF-alpha on SWO glioma cell proliferation with or without U73122 treatment was investigated by MTT assay. The cell apoptosis induced by TNF-alpha along or in combination with U73122 was detected by flow cytometry with PI staining. The expression of caspase-3 and Bcl-2 was detected by Western blotting. RESULTS AND CONCLUSION: U73122 can sensitize SWO glioma cells to TNF-alpha-induced apoptosis. Blocking the PLC-gamma1 pathway may not induce apoptosis of SWO glioma cells, but can sensitize SWO glioma cells to small-dose TNF-alpha-induced apoptosis, the mechanism of which may involve down-regulation of bcl-2.

Apoptosis↗

[Extraction and determination of short-chain fatty acids in biological samples].

Short-chain fatty acids are organic acids with 1 - 6 carbon atoms. Their physiological functions and clinical applications have attracted considerable attention. Their high polarity, low ultraviolet (UV) absorbance, low contents in biological samples, high volatility, good water-solubility and easy adsorption on metal and glass surfaces result in some difficulties in their separation and determination. This review focuses on the methods for extraction of short-chain fatty acids in biological samples such as feces, urine, blood and culture solutions by distillation, high speed centrifugation, liquid-solid extraction, solid phase microextration, supercritical fluid extraction and liquid phase microextraction and determination by gas chromatography, high performance liquid chromatography and capillary electrophoresis. Sixty-three references are cited.

Adsorption↗

Effect of the combination of fibrin glue and growth hormone on incomplete intestinal anastomoses in a rat model of intra-abdominal sepsis.

BACKGROUND: The presence of established intra-abdominal sepsis has been considered a contraindication to primary anastomoses. Our hypothesis was that fibrin glue (FG), growth hormone (rhGH), and combination of them synergistically improve intestinal primary anastomotic healing in a rat model of intestinal fistulae with peritonitis. MATERIALS AND METHODS: Male Wistar rats, induced intestinal fistulae with peritonitis after 24 h, were performed an enterectomy and intestinal anastomoses. Group A, rats (n = 60) had a complete anastomoses (end-to-end single layer anastomoses using 12 inverted interrupted 6-0 sutures) without peritonitis, group B, rats (n = 60) had a complete anastomoses after 24 h of peritonitis, group C rats had an incomplete anastomoses (four inverted interrupted sutures), groups D, E, F rats (n = 60) received FG, rhGH, or both of them, respectively. rhGH was given daily for 5 days. Anastomoses indicated the anastomotic bursting pressure (ABP), tensile strength, and hydroxyproline content, were determined. RESULTS: On POD 1, ABP of group C and group D was significantly lower than that of other groups (P < 0.01); On POD 3, ABP could not be determined because of intestinal dehiscence in groups C and E, ABP was significantly higher in groups D and F than that of groups A and B (P < 0.01); the ABP increased after 5 days of operation in groups A, B, and F. At the same time, that of group D decreased (P < 0.01). On POD 5, the tensile strength was significantly higher in groups A, D, and F than that in groups C, and E. On POD 5, hydroxyproline content was higher in groups D and F compared to that in group C (P < 0.05). CONCLUSIONS: These data suggested that FG improve intestinal primary anastomotic healing within post-operative 5 days in a rat model of intestinal fistulae with peritonitis. RhGH alone fails to improve intestinal anastomotic healing, and the combination of FG and rhGH have no synergistic effect to improves intestinal anastomotic healing.

Anastomosis, Surgical↗

Molecular beacons for bioanalytical applications.

Molecular beacons (MBs) are hairpin-shaped oligonucleotides that contain both fluorophore and quencher moieties. They act like switches and are normally in a closed state, when the fluorophore and the quencher are brought together to turn "off" the fluorescence. When prompted to undergo conformational changes that open the hairpin structure, the fluorophore and the quencher are separated, and fluorescence is turned "on." This Education will outline the principles of MBs and discuss recent bioanalytical applications of these probes for in vitro RNA and DNA monitoring, biosensors and biochips, real-time monitoring of genes and gene expression in living systems, as well as the next generation of MBs for studies on proteins, the MB aptamers. These important applications have shown that MBs hold great potential in genomics and proteomics where real-time molecular recognition with high sensitivity and excellent specificity is critical.

Biosensing Techniques↗

Docosahexaenoic acid changes lipid composition and interleukin-2 receptor signaling in membrane rafts.

Polyunsaturated fatty acids, including docosahexaenoic acid (DHA, 22:6n-3), modulate immune responses and exert beneficial immunosuppressive effects, but the molecular mechanisms inhibiting T-cell activation are not yet elucidated. Lipid rafts have been shown to play an important role in the compartmentalization and modulation of cell signaling. We investigated the role of DHA in modulating the lipid composition in lipid rafts and membrane subdomain distribution of interleukin-2 (IL-2) receptor signaling molecules. We found that DHA altered lipid components of rafts and modified the IL-2-induced Janus kinase-signal transducer and activator of transcription (STAT) signaling pathway by partially displacing IL-2 receptors from lipid rafts. We fractionated plasma membrane subcellular compartments and discovered that certain amounts of STAT5a and STAT5b existed in detergent-resistant plasma membrane fractions of T-cells. After DHA treatment, STAT5a and STAT5b were not detected in lipid raft fractions and were located in detergent-soluble fractions. These data demonstrate for the first time that DHA alters the lipid composition of membrane microdomains and suppresses IL-2 receptor signaling in T-cells. Thus, our data provide evidence for a functional modification in lipid rafts by DHA treatment and explain PUFA-mediated immunosuppressive effects.

Cell Membrane↗

Wortmannin induces zebrafish cardia bifida through a mechanism independent of phosphoinositide 3-kinase and myosin light chain kinase.

Cardia bifida is an anomaly of the embryonic heart in which the bilateral myocardial rudiments fail to travel to the midline, resulting in the formation of two separate hearts in lateral positions. In zebrafish, eight loci responsible for the cardia bifida phenotype were identified in the large-scale genetic screen. Wortmannin has been reported to be a highly selective inhibitor of phosphoinositide 3-kinase and myosin light chain kinase activity. We provide the first evidence that wortmannin treatment of zebrafish embryos can induce cardia bifida in a dose-dependent manner and that wortmannin alters cardiac development between 6 and 16 h post-fertilization. In addition, we demonstrate that wortmannin induces zebrafish cardia bifida through a mechanism independent of phosphoinositide 3-kinase and myosin light chain kinase. Our findings may provide new insights into the cardiomyocyte function and disfunction.

Androstadienes↗

Simultaneous determination of l- and d-lactic acid in plasma by capillary electrophoresis.

A novel method for simultaneous determination of d- and l-lactic acids in plasma was presented by capillary electrophoresis with photodiode array detection at 195nm. The separation was performed in an uncoated fused-silica capillary. The parameters influencing the resolution and the migration time of lactic acids were optimized. When 150mM phosphate-Tris buffer (pH 7.0) consisting of 220mM 2-hydroxypropyl-beta-cyclodextrin and 0.2mM tetradecyltrimethylammonium bromide was utilized as the running buffer, highly effective chiral separation of d- and l-lactic acids was achieved at about 42min at an effective voltage of -25kV. The resolution of lactic acid enantiomers was >/=1.25. The limits of detection of d- and l-lactic acids in standard solution without any pretreatment were 80 and 50muM (S/N=3), respectively. Sample pretreatment was preceded by protein-removal procedure with acetonitrile. With a pre-concentration procedure by 10 times, the limits of detection of d- and l-lactic acids were 20 and 15muM (S/N=10), respectively. The satisfactory analytical performance of the proposed method was validated.

Animals↗

Role of the pentanucleotide (tttta)n polymorphisms of Cyp11alpha gene in the pathogenesis of hyperandrogenism in Chinese women with polycystic ovary syndrome.

To determine the (tttta)n repeat polymorphisms at the promoter region of CYP11alpha gene, and study its linkage to hyperandrogenism of polycystic ovary syndrome (PCOS) in Chinese women, a case-control study was conducted in the Reproductive Medical Center of the Second Affiliated Hospital of Zhengzhou University (Zhengzhou, China). 96 PCOS patients and 78 healthy control women were included. CYP11alpha (tttta)n repeat-polymorphism genotyping analysis was performed by using polymerase chain reaction (PCR). Serum pituitary hormone and total testosterone levels were measured by ELISA. 4 different CYP11alpha (tttta)n allelles were identified, corresponding to 4-, 6-, 8-, and 9-repeat-unit alleles. The frequency and distribution of these alleles are 0.16, 0.33, 0.38, and 0.13 respectively in PCOS patients, as compared with 0.20, 0.34, 0.35, and 0.11 respectively in healthy controls. There were no significant differences between these two groups. Moreover, no correlation between the polymorphism of CYP11alpha gene and serum testosterone level of patients with PCOS and controls was observed. It is concluded that microsatellite polymorphism (tttta)n of gene CYP11alpha exists in Chinese women and the polymorphism of CYP11alpha gene does not play an important role in the pathogenesis of Chinese patients with PCOS, especially in patients with hyperandrogenism.

Adult↗

Identification of a novel nucleus protein involved in the regulation of urokinase in 95D cells.

The urokinase-type plasminogen activator (uPA) plays an important role in cellular invasion. By using the downstream part of a 74 bp DNA region called the cooperation mediator (COM) of the uPA promoter as a bait sequence in the yeast one-hybrid screen, a gene called PBK1 was previously cloned from the cDNA library of the 95D lung cancer cell strain. In this study, the intracellular distribution of PBK1 was studied by using the transient transfection of pEGFP-C3-PBK1, and PBK1 was found to be localized in the nucleus. Co-transfection of pEGFP-C3-PBK1 and the deletion mutants of the pGL3-uPA promoter indicated that PBK1 can increase the uPA promoter activity by about 25% and this effect is uPA enhancer-dependent. Western blotting and Enzyme-linked immunoadsordent assay further confirmed that PBK1 can upregulate the expression of uPA. Our results suggest that PBK1 is involved in the regulation of uPA expression, which might provide a new clue to further understanding the regulation mechanism of uPA expression.

Cell Line, Tumor↗

Identification of gmhA, a Yersinia pestis gene required for flea blockage, by using a Caenorhabditis elegans biofilm system.

Yersinia pestis, the cause of bubonic plague, blocks feeding by its vector, the flea. Recent evidence indicates that blockage is mediated by an in vivo biofilm. Y. pestis and the closely related Yersinia pseudotuberculosis also make biofilms on the cuticle of the nematode Caenorhabditis elegans, which block this laboratory animal's feeding. Random screening of Y. pseudotuberculosis transposon insertion mutants with a C. elegans biofilm assay identified gmhA as a gene required for normal biofilms. gmhA encodes phosphoheptose isomerase, an enzyme required for synthesis of heptose, a conserved component of lipopolysaccharide and lipooligosaccharide. A Y. pestis gmhA mutant was constructed and was severely defective for C. elegans biofilm formation and for flea blockage but only moderately defective in an in vitro biofilm assay. These results validate use of the C. elegans biofilm system to identify genes and pathways involved in Y. pestis flea blockage.

Animals↗

Yersinia pestis is viable with endotoxin composed of only lipid A.

Lipopolysaccharide (LPS) is the major outer membrane component of gram-negative bacteria. The minimal LPS structure for viability of Escherichia coli and Salmonella enterica serovar Typhimurium is lipid A glycosylated with 3-deoxy-D-manno-octulosonic acid (Kdo) residues. Here we show that another member of the Enterobacteriaceae, Yersinia pestis, can survive without Kdo in its LPS.

Bacterial Toxins↗

[Relationship between the microsatellite polymorphism of CYP11 alpha gene and the pathogenesis of hyperandrogenism of polycystic ovary syndrome in Chinese].

OBJECTIVE: To explore the association between the microsatellite polymorphism in the promoter region of CYP11 alpha gene and hyperandrogenism of polycystic ovary syndrome (PCOS). METHODS: Eighty-six cases of PCOS and 50 normal women as controls were studied. Polymerase chain reaction and electrophoresis on polyacrylamide gel were employed to detect the polymorphism of CYP11 alpha gene and its frequency distribution. At the same time, the relationships of CYP11 alpha alleles to serum testosterone levels in PCOS were compared. RESULTS: Four different CYP11 alpha (tttta)n alleles were identified, corresponding to 4, 6, 8 and 9 repeat-units alleles. The frequency distribution profiles were 0.17, 0.31, 0.39, 0.13 and 0.22, 0.35, 0.33, 0.10 in PCOS group and control group respectively, showing no statistically significant difference between the two groups. There were no correlations between the polymorphism of CYP11 alpha gene and the serum testosterone levels of PCOS patients. CONCLUSION: Micro-satellite polymorphism (tttta)n of gene CYP11 alpha exists in Chinese women and the polymorphism does not relate to the pathogenesis of hyperandrogenism in women with PCOS.

Adult↗