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X Ji

Publications and source records attributed to X Ji.

At least 37 records · Page 2Linked to original sources

Crystal structure of unligated guanylate kinase from yeast reveals GMP-induced conformational changes.

The crystal structure of guanylate kinase (GK) from yeast (Saccharomyces cerevisiae) with a non-acetylated N terminus has been determined in its unligated form (apo-GK) as well as in complex with GMP (GK.GMP). The structure of apo-GK was solved with multiwavelength anomalous diffraction data and refined to an R-factor of 0.164 (R(free)=0.199) at 2.3 A resolution. The structure of GK.GMP was determined using the crystal structure of GK with an acetylated N terminus as the search model and refined to an R-factor of 0.156 (R(free)=0.245) at 1.9 A. GK belongs to the family of nucleoside monophosphate (NMP) kinases and catalyzes the reversible phosphoryl transfer from ATP to GMP. Like other NMP kinases, GK consists of three dynamic domains: the CORE, LID, and NMP-binding domains. Dramatic movements of the GMP-binding domain and smaller but significant movements of the LID domain have been revealed by comparing the structures of apo-GK and GK.GMP. apo-GK has a much more open conformation than the GK.GMP complex. Systematic analysis of the domain movements using the program DynDom shows that the large movements of the GMP-binding domain involve a rotation around an effective hinge axis approximately parallel with helix 3, which connects the GMP-binding and CORE domains. The C-terminal portion of helix 3, which connects to the CORE domain, has strikingly higher temperature factors in GK.GMP than in apo-GK, indicating that these residues become more mobile upon GMP binding. The results suggest that helix 3 plays an important role in domain movement. Unlike the GMP-binding domain, which moves toward the active center of the enzyme upon GMP binding, the LID domain moves away from the active center and makes the presumed ATP-binding site more open. Therefore, the LID domain movement may facilitate the binding of MgATP. The structure of the recombinant GK.GMP complex superimposes very well with that of the native GK.GMP complex, indicating that N-terminal acetylation does not have significant impact on the three-dimensional structure of GK.

Acetylation↗

[3H]MRS 1754, a selective antagonist radioligand for A(2B) adenosine receptors.

MRS 1754 [N-(4-cyanophenyl)-2-[4-(2,3,6,7-tetrahydro-2,6-dioxo-1,3-dipropyl-1H-purin-8-yl)-phenoxy]acetamide] is a selective antagonist ligand of A(2B) adenosine receptors. This is the least well-defined adenosine receptor subtype, and A(2B) antagonists have potential as antiasthmatic drugs. For use as a radioligand, MRS 1754, a p-cyanoanilide xanthine derivative, was tritiated on the propyl groups in a two-step reaction using a p-carboxamido precursor, which was dehydrated to the cyano species using trifluoroacetic anhydride. [3H]MRS 1754 (150 Ci/mmol) bound to recombinant human A(2B) adenosine receptors in membranes of stably transfected HEK-293 cells. Specific binding was saturable, competitive, and followed a one-site model, with a K(D) value of 1.13 +/- 0.12 nM and a B(max) value of 10.9 +/- 0.6 pmol/mg protein. Specific binding utilizing 0.7 nM [3H]MRS 1754 was > 70% of total binding. The affinity calculated from association and dissociation binding constants was 1.22 nM (N = 4). Binding to membranes expressing rat and human A(1) and A(3) adenosine receptors was not significant, and binding in membranes of HEK-293 cells expressing human A(2A) receptors was of low affinity (K(D) > 50 nM). The effects of cations and chelators were explored. Specific binding was constant over a pH range of 4.5 to 6.5, with reduced binding at higher pH. The pharmacological profile in competition experiments with [3H]MRS 1754 was consistent with the structure-activity relationship for agonists and antagonists at A(2B) receptors. The K(i) values of XAC (xanthine amine congener) and CPX (8-cyclopentyl-1,3-dipropylxanthine) were 16 and 55 nM, respectively. NECA (5'-N-ethylcarboxamidoadenosine) competed for [3H]MRS 1754 binding with a K(i) of 570 nM, similar to its potency in functional assays. Thus, [3H]MRS 1754 is suitable as a selective, high-affinity radioligand for A(2B) receptors.

Acetamides↗

Studying hadronic structure of the photon in lattice QCD.

We show that the matrix element of a local quark-gluon operator in the photon state, , can be calculated in lattice QCD. The result is generalized to other quantities involving spacelike photons, including the transition form factor gammagamma*-->pi(0) and the virtual-photon-nucleon Compton amplitude which can be used to define the generalized Drell-Hearn-Gerasimov and Bjorken sum rules.

Journal Article↗

International Child Care Practices Study: infant sleeping environment.

BACKGROUND: The International Child Care Practices Study (ICCPS) has collected descriptive data from 21 centres in 17 countries. In this report, data are presented on the infant sleeping environment with the main focus being sudden infant death syndrome (SIDS) risk factors (bedsharing and infant using a pillow) and protective factors (infant sharing a room with adult) that are not yet well established in the literature. METHODS: Using a standardised protocol, parents of infants were surveyed at birth by interview and at 3 months of age mainly by postal questionnaire. Centres were grouped according to geographic location. Also indicated was the level of SIDS awareness in the community, i.e. whether any campaigns or messages to "reduce the risks of SIDS" were available at the time of the survey. RESULTS: Birth interview data were available for 5488 individual families and 4656 (85%) returned questionnaires at 3 months. Rates of bedsharing varied considerably (2-88%) and it appeared to be more common in the samples with a lower awareness of SIDS, but not necessarily a high SIDS rate. Countries with higher rates of bedsharing appeared to have a greater proportion of infants bedsharing for a longer duration (>5 h). Rates of room sharing varied (58-100%) with some of the lowest rates noted in centres with a higher awareness of SIDS. Rates of pillow use ranged from 4% to 95%. CONCLUSIONS: It is likely that methods of bedsharing differ cross-culturally, and although further details were sought on different bedsharing practices, it was not possible to build up a composite picture of "typical" bedsharing practices in these different communities. These data highlight interesting patterns in child care in these diverse populations. Although these results should not be used to imply that any particular child care practice either increases or decreases the risk of SIDS, these findings should help to inject caution into the process of developing SIDS prevention campaigns for non-Western cultures.

Beds↗

Crystallographic and modeling studies of RNase III suggest a mechanism for double-stranded RNA cleavage.

BACKGROUND: Aquifex aeolicus Ribonuclease III (Aa-RNase III) belongs to the family of Mg(2+)-dependent endonucleases that show specificity for double-stranded RNA (dsRNA). RNase III is conserved in all known bacteria and eukaryotes and has 1-2 copies of a 9-residue consensus sequence, known as the RNase III signature motif. The bacterial RNase III proteins are the simplest, consisting of two domains: an N-terminal endonuclease domain, followed by a double-stranded RNA binding domain (dsRBD). The three-dimensional structure of the dsRBD in Escherichia coli RNase III has been elucidated; no structural information is available for the endonuclease domain of any RNase III. RESULTS: We present the crystal structures of the Aa-RNase III endonuclease domain in its ligand-free form and in complex with Mn(2+). The structures reveal a novel protein fold and suggest a mechanism for dsRNA cleavage. On the basis of structural, genetic, and biological data, we have constructed a hypothetical model of Aa-RNase III in complex with dsRNA and Mg(2+) ion, which provides the first glimpse of RNase III in action. CONCLUSIONS: The functional Aa-RNase III dimer is formed via mainly hydrophobic interactions, including a "ball-and-socket" junction that ensures accurate alignment of the two monomers. The fold of the polypeptide chain and its dimerization create a valley with two compound active centers at each end of the valley. The valley can accommodate a dsRNA substrate. Mn(2+) binding has significant impact on crystal packing, intermolecular interactions, thermal stability, and the formation of two RNA-cutting sites within each compound active center.

Amino Acid Sequence↗

Structure and dynamics of 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase.

Folates are essential for life. Unlike mammals, most microorganisms must synthesize folates de novo. 6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) catalyzes pyrophosphoryl transfer from ATP to 6-hydroxymethyl-7,8-dihydropterin (HP), the first reaction in folate pathway, and therefore, is an ideal target for developing novel antimicrobial agents. Because of its small size and high thermal stability, E. coli HPPK is also an excellent model enzyme for studying the mechanisms of enzymatic pyrophosphoryl transfer. We have determined the crystal structures of HPPK in the unligated form and in complex with HP, two Mg2+ ions, and AMPCPP (an ATP analog that inhibits the enzymatic reaction). Comparison of the two crystal structures reveals dramatic conformational changes of three flexible loops and many side chains and possible roles of the active site residues.

Adenosine Triphosphate↗

The effects of thermal and hydric environments on hatching success, embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata.

We examined the effects of thermal and hydric environments on hatching success, the embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata. The eggs were incubated at four temperatures ranging from 24 to 32 degrees C on substrates with water potentials of 0 and -220 kPa using a 4x2 factorial design. Both thermal and hydric environments affected the water exchange between eggs and their surroundings. Eggs incubated in wetter substrates gained mass throughout the course of incubation, whereas eggs in drier substrates gained mass during the first half of incubation and lost mass thereafter. Hatching success was noticeably higher at 26 and 30 degrees C than at 24 and 32 degrees C, but among treatments, differences in hatching success were not significant. Temperature significantly affected the duration of incubation and most hatchling traits examined. Deformed hatchlings were found in all temperature treatments, with more deformities observed at 32 degrees C. Hatchlings from eggs incubated at different temperatures differed in wet body mass, but the differences stemmed mainly from variation in water contents. Embryos at different temperatures completed development at nearly the same expenditure of energy and catabolized nearly the same amount of lipids, but hatchlings from different temperatures differed in the development condition of carcass at hatching. Hatchlings from eggs incubated at 26 degrees C were larger in SVL than those from other higher or lower incubation temperatures, characteristically having larger carcasses; hatchlings from 32 degrees C eggs were smaller in SVL and had smaller carcasses but larger residual yolks than those from lower incubation temperatures. Hatchlings from eggs incubated at 24 degrees C were shorter in tail length but greater in size (SVL)-specific body wet mass than those from higher incubation temperatures. Within the range from -220 to 0 kPa, the substrate water potential did not affect hatching success, the embryonic use of energy and all hatchling traits examined, and the effects of temperature were independent of the effects of substrate water potential. Therefore, our data add evidence showing that embryonic development in reptiles with pliable-shelled eggs is relatively insensitive to variation in hydric environments during incubation.

Animals↗

Role of arginine 216 in catalytic activity of murine Alpha class glutathione transferases mGSTAl-1 and mGSTA2-2 toward carcinogenic diol epoxides of polycyclic aromatic hydrocarbons.

Murine class Alpha glutathione (GSH) transferase A1-1 (mGSTA1-1) is unique among mammalian Alpha class GSTs due to its exceptionally high catalytic activity toward (+)-anti-7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [(+)-anti-BPDE], which is the activated metabolite of an environmentally relevant carcinogen, benzo[a] pyrene (BP). However, the molecular basis for high catalytic activity of mGSTA1-1 toward (+)-anti-BPDE is not clear. In the present study, we demonstrate that an arginine residue at position 216, which is conserved in some but not all mammalian class Alpha GSTs, plays an important role in catalytic activity of mGSTA1-1 toward (+)-anti-BPDE and carcinogenic diol epoxides of other environmentally relevant polycyclic aromatic hydrocarbons (PAHs). The catalytic efficiency (k(cat)/K(m)) of mGSTA1-1 for the GSH conjugation of (+)-anti-BPDE (108/mM/s) was reduced by about 58% upon replacement of arginine 216 with alanine (R216A). This was mainly due to a significantly lower V(max) for the R216A mutant of mGSTA1-1 compared with wild-type mGSTA1-1. The R216A mutation also resulted in a statistically significant reduction (>70%) in specific activity of mGSTA1-1 toward racemic anti-diol epoxides of chrysene and benzo[c]phenanthrene (anti-CDE and anti-B[c]PDE, respectively). The catalytic activity of mGSTA2-2, which is a close structural homologue of mGSTA1-1, was also reduced upon R216A mutation. The results of the present study clearly indicate that an arginine residue at position 216 is critical for catalytic activity of mGSTA1-1 and mGSTA2-2 toward carcinogenic diol epoxide metabolites of various PAHs that are abundant in the environment and suspected human carcinogens.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

[Optimization analysis of ventilation and dehumidification in manned spacecraft].

OBJECTIVE: To achieve the optimized design of system operational parameters and system components in air ventilation and dehumidification of manned spacecraft. METHOD: Mathematical-physical model of air ventilation and dehumidification system was established. Through the design of optimal operational parameters for system performance, heat and moisture in the cabin of manned spacecraft could be removed effectively, dewing could be prevented, and the comfort demand of the astronauts could be met. Equivalent mass of the system in different operational parameters and system components was analyzed by calculation. RESULT: The dew point temperature and the comfortability depended on the temperature and humidity of the influx, ventilation rate, and evaporation of moisture in the cabin. There were two ways to control the operational parameters of air ventilation and dehumidification system. The method based on controlling temperature didn't use recuperative heat exchangers in the loop, and the system structure for the design was simple, but it didn't meet the requirement of dissipating moisture in some conditions. The other method was based on controlling the moisture, and it could steadily keep the dew point temperature in the cabin and control the occurrence of dew. But the system structure of this design was relatively complex. CONCLUSION: Mass and power consumption of the system could be reduced by selecting appropriate parameters and adopting the measures of optimization.

Air Conditioning↗

[The transcription of interleukin-8 in gastric epithelial cells by Helicobacter pylori strains in Chinese].

OBJECTIVE: To observe the effect of the vac A genotype and the existence of cag pathogenicity island (cag PAI) of Helicobacter pylori (Hp) isolated from Chinese patients on the transcription of IL-8 in gastric epithelial cells. METHODS: The genotype of vac A and the existence of cag PAI of Hp were determined by gene sequencing and Southern blot, respectively; the constructed L5F11 cells were cocultured with the bacteria and the transcription of interleukin-8(IL-8) was indicated by luciferase activity and the cytotoxin activity showed by using HeLa cell vacuolating assay. RESULTS: The ability to induce luciferase by the strains with complete cag PAI (n = 15) was much higher than that by the wild cag negative strain G50[(0.47 +/- 0.09) x 10(6) cpm vs (0.13 +/- 0.02) x 10(6) cpm, P < 0.01] and there was no difference in the luciferase activity induced by the strains with partial cag (n = 4) and by G50[(0.23 +/- 0.08) x 10(6) cpm vs (0.13 +/- 0.02) x 10(6) cpm, P > 0.05]; Also no difference was found in the luciferase activity induced by the strains with vac A s1/m1 (n = 5) and the strains with s1/m2 (n = 14) [(0.29 +/- 0.12) x 10(6) cpm vs (0.53 +/- 0.41) x 10(6) cpm, P > 0.05] although the formers had much higher vacuolating cytotoxin activity than the latters (P < 0.001). CONCLUSION: The transcription of IL-8 in gastric epithelial cells induced by Hp isolated from Chinese patients was associated with the existence of cag PAI and not with the genotype of vac A.

Bacterial Proteins↗

[Effect of environmental factors on N and P uptake by Gracilaria tenuistipitata var. liui Zhang et Xia].

The effect of light intensity, pH, temperature, salinity on the uptake of nitrogen and phosphorus by Gracilaria tenuistipitata var. liui Zhang et Xia was were studied under laboratory conditions. The uptake rate of nitrogen reached as high as 0.32-0.36 mumol.g-1.h-1 at favorable light intensity of 800-2400 mumol photon.m-2.s-1, while that of phosphorus reached 0.017-0.018 mumol.g-1.h-1 at 320-1600 mumol photon.m-2.s-1. The nutrient uptake rate reached the maximum at pH 8.0. A significant difference of N uptake rate at different temperatures or salinities and that of P uptake rate at different temperatures were found. The N uptake rate was higher at 21-26 degrees C and 20@1000-28@1000, but P uptake was higher at 21-26 degrees C. There was no interaction between temperature and salinity.

Absorption↗

[The clinical effect of non-penetrating trabecular surgery with reticulated sodium hyaluronate implant for treatment of primary open-angle glaucoma].

OBJECTIVE: To evaluate the clinical effect of non-penetrating trabecular surgery (NPTS) with reticulated sodium hyaluronate implant in primary open-angle glaucoma (POAG). METHODS: NPTS with reticulated sodium hyaluronate implant was performed on 27 eyes of 25 patients with POAG. The procedure consisted of excising a deep scleral tissue including the external wall of the Schlemm's canal under a scleral flap without opening the anterior chamber, as the inner wall of the canal was left in place. Then placing a 3.0 mm x 4.5 mm x 0.5 mm or 3.5 mm x 3.5 mm x 3.5 mm reticulated sodium hyaluronate implant was placed under the flap, so that the aqueous humor may filter through the thin layer of trabeculocorneal membrane spontaneously without forming a flat chamber. The postoperative intraocular pressure (IOP), inflammation and filtering bleb were analyzed. All of patients undertook ultrasound biomicroscopy (UBM) and gonioscopy to evaluate the surgical site on the postoperative 2 weeks, 3, 6 and 9 months. The mean follow-up was (6.61 plus minus 2.47) months. RESULTS: The IOP decreased from a mean preoperative value of (26.36 +/- 9.02) mm Hg (1 mm Hg = 0.133 kPa) to a mean postoperative value of (14.18 +/- 3.51) mm Hg (t = 6.875, P < 0.05). The number of anti-glaucomatous medications, topical or systemical, was reduced from (2.96 +/- 1.43) sorts preoperatively to (0.77 +/- 1.07) sorts postoperatively. The visual acuity remained stable (no statistical difference with chi(2) test). Six eyes had elevated IOP after operation and were controlled by eyedrops or trabecular puncture with ND: YAG laser. A slight hyphema occurred in 4 eyes with small puncture during operation. The complications such as flat chamber, inflammation and choroidal detachment were not observed. The UBM showed that the sodium hyaluronate implant degraded and a transparent liquid space existed under the scleral flap in all of patients at postoperative 3 months. The gonioscopy showed that at the surgical site the thinner and semitransparent trabecular membrane and changes of a transparent cavity could be seen in 23 eyes. In the other eyes, at the site the trabecula was translucent. The filtering blebs were formed in 8 eyes. CONCLUSION: Non-penetrating trabecular surgery with reticulated sodium hyaluronate implant can effectively lower the IOP and reduce the sorts of anti-glaucomatous medications. The visual acuity may remain unchanged, and no serious complications commonly seen in the traditional trabeculectomy occur. The procedure is a new simple and effective one for the treatment of POAG.

Adolescent↗

[Gene rearrangement and p53 expression in defining the nature of angioimmunoblastic lymphadenopathy].

OBJECTIVE: To investigate gene rearrangement and p53 expression in defining the nature of angioimmunoblastic lymphadenopathy. METHODS: DNA was extracted from paraffin-embedded tissue blocks of 44 angioimmunoblastic lymphadenopathy (AIL) patients and analyzed with polymerase chain reaction (PCR) for IgH and TCRgamma gene rearrangement. Immunohistochemistry staining was used to detect p53 protein expression. Thirty-five cases were followed-up. RESULTS: 12 out of 44 cases (27.3%) showed TCRgamma gene rearrangement and 2 (4.5%) showed IgH gene rearrangement. Rearrangement of both IgH and TCRgamma genes were detected in 2 cases (4.5%). 14 cases (31.8%) showed p53 positive expression, among which 12 showed positive rearrangement and 2 showed negative (P < 0.01). Eight out of 11 patients of positive gene rearrangement died in one year, while only 3 patients were still alive at the eighteenth month of follow-up, three of 24 patients of negative gene rearrangement were found dead at the time of the one year follow-up, while the rest 21 patients were alive and the longest survival time was 96 months. CONCLUSIONS: Gene rearrangement can define the pathological nature of AIL. The expression of p53 is highly related to gene rearrangement, and thus an important immunological marker in research on AIL.

Follow-Up Studies↗

[Treatment of nuclein 32P colloid perfusion with chronic suppurative maxillary sinusitis].

OBJECTIVE: To determine the effect of nuclein 32P colloid in treatment of chronic maxillary sinusitis (CMS). METHOD: 22 patients (31 sides) with CMS after washing of sinucavity were perfused with nuclein 32P in sinucavity. 20 patients (31 sides) with routine conservative treatment were served as a control group. Two groups were followed up for one year. RESULT: In nuclein group, 27 sides were cured, 4 sides were recurred and the rate of cure was 87%. In routine group, 19 sides were cured, 12 sides were recurred and the rate of cure is 61%. There is significant difference between the two groups (P < 0.01). The treatment active of nuclein group is better than that of routine group. CONCLUSION: Nuclein 32P colloid with the function of ionizing radiation can increase the pathogenic tissue immunocompetence and metabolic course, obstruct the microvascular and promote the absorption of inflammatory exudation and edema subsidence. It can also make the proliferous tissue atrophy thence accelerate the rate of cure of CMS.

Adult↗

[A study on the effect of rare earth metal ions on fluorescence spectra of the tryptophan using fluorescence spectroscopy].

Rare earth metal ions and tryptophan form ion-association complex in basic medium. The complex causes fluorescence quenching of tryptophan. Fluorescence emission of tryptophan and quenching caused by rare earth metal ions both reach a climax in H3BO4-HAc-H3PO4-NaOH at pH 10 to 11, and all rare earth metal ions have the approximate effects on fluorescence quenching of tryptophan. The molecular mode of complex of rare earth metal ions and tryptophan has been founded and mechanism of fluorescence quenching has been studied in this paper.

Chelating Agents↗

[The role of cell junction in hypothermic impairment of tissue].

OBJECTIVE: To investigate the role of cell junction in the hypothermal impairment of tissue without any cryoprotectant. METHODS: Epithelial MDCK cell line which is able to form well-defined cell junctions after confluency, and fibroblast V79 cell line which is unable to be used for graded freezing experiments, either in suspension, single tough attached to flasks or as monolayers on flasks. Then the viability was assessed by WST-1 and MT, respectively. RESULTS: The results of viability of the two types of cells assessed by WST-1 were well-fitted with that of MTT. The recovery of the two types of cells in suspension was higher than that of single tough group and monolayer group. The single tough group of MDCK had a higher recovery than did the monolayer group. At the same time, such tendency was not observed on V79 cells. As compared with V79 monolayers, lower recovery was found on MDCK monolayers. CONCLUSIONS: The junction between cells and the adhesion between cell and matrix have important roles in the hypothermic impairment of tissue. Their existence affect the recovery of cells under hypothermia. The cytoskeleton system may contribute to the impairment caused by junctions.

Animals↗

Complete crystal structure of monocyte chemotactic protein-2, a CC chemokine that interacts with multiple receptors.

Monocyte chemotactic protein 2 (MCP-2) is a CC chemokine that utilizes multiple cellular receptors to attract and activate human leukocytes. MCP-2 is a potent inhibitor of HIV-1 by virtue of its high-affinity binding to the receptor CCR5, one of the major coreceptors for HIV-1. Although a few structures of CC chemokines have been reported, none of these was determined with the N-terminal pyroglutamic acid residue (pGlu1) and a complete C-terminus. pGlu1 is essential for the chemotactic activity of MCP-2. Recombinant MCP-2 has Gln1 at the N terminus, 12-15% of which cyclizes automatically and forms pGlu1. The chemotactic activity of such MCP-2 mixture, which contains 12-15% pGlu1-form and 85-88% Gln1-form protein, is approximately 10 times lower when compared with that of fully cyclized MCP-2 preparation. Therefore, this chemokine is practically inactive without pGlu1. We have determined the complete crystal structure of MCP-2 that contains both pGlu1 and an intact C-terminus. With the existence of pGlu1, the conformation of the N-terminus allows two additional interactions between the two subunits of MCP-2 dimer: a hydrogen bond between pGlu1 and Asn17 and a salt bridge between Asp3 and Arg18. Consequently, both pGlu1 are anchored and buried, and thereby, both N-terminal regions are protected against protease degradation. We have also observed not previously reported extended helical nature of the C terminal region, which covers residues 58-74.

Amino Acid Sequence↗