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Biomedical subjects

Jun Wu

Publications and source records attributed to Jun Wu.

At least 109 records · Page 6Linked to original sources

CTLA4Ig introduced by adenovirus vector locally to prolong the survival of xenogeneic skin grafts on rat burn wounds.

BACKGROUND: The purpose of this study was to explore an applicable approach for prolonging the survival of heterogenetic skin grafts on burn wounds with CTLA4Ig. METHODS: An adenovirus vector named Ad-CTLA4Ig, which could express human CTLA4Ig fusion protein, was constructed. Infecting and replicating in 293 cells, more Ad-CTLA4Ig and recombinant human CTLA4Ig (rhCTLA4Ig) were prepared, respectively. In a rat flame thermal injury model, the effect of rhCTLA4Ig on survival time of human skin graft on the eschar-excised rat burn wound was observed. Meanwhile, the efficiency of Ad-CTLA4Ig infecting cultured skin fibroblasts, keratinocytes, and partial-thickness skin samples were checked by CTLA4Ig expression essay. Then, the Ad-CTLA4Ig was administered locally on the eschar-excised wound and dermis of the skin graft, and the survival time of the human skin graft on burn wound was measured. The influence of the systemic immune function by rhCTLA4Ig and Ad-CTLA4Ig were also determined. RESULTS: The prepared rhCTLA4Ig from the supernatant of Ad-CTLA4Ig-infected 293 cells was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis sodium dodecyl sulfate poly-acrylamide gel electrophoresis and Western blot. It was found that CTLA4Ig could significantly prolong the xenogeneic skin graft survival in a dosage-dependent manner. Interestingly, the survival time was longer when CTLA4Ig was used 24 hours posttransplantation than that at hour 0. The expression of CTLA4Ig could be observed in the cultured skin fibroblasts, keratinocytes, and skin pieces soon after Ad-CTLA4Ig transfection, as demonstrated by either immunocellular chemistry or immunohistochemistry assay. When Ad-CTLA4Ig was locally administered during skin transplantation on burn wound, the survival time was increased from 7.9 days of control group to 21.6 days, whereas the systemic immune function was not affected. CONCLUSION: Administration of Ad-CTLA4Ig locally could prolong the survival time of xenogeneic skin graft on burn wound without significantly influencing the systemic immune function.

Abatacept↗

A new unsaturated glycoglycerolipid from a cultured marine dinoflagellate Amphidinium carterae.

From the cultured marine dinoflagellate Amphidinium carterae, a new unsaturated glycoglycerolipid (2S)-1,2-O-6,9,12,15-dioctadecatetraenoyl-3-O-[alpha-D-galactopyranosyl-(1''''-->6''')-O-beta-D-galactopyranosyl]-glycerol (1), has been isolated together with two known saturated ones, (2S)-1,2-distearoyl-3-O-(6-sulpho-alpha-D-quinovopyranosyl)-glycerol (2) and (2S)-1-stearoyl-3-O-(6-sulpho-alpha-D-quinovopyranosyl)-glycerol (3). Their structures were elucidated on the basis of chemical and spectral data.

Animals↗

[Construction and identification of recombinant adenovirus vector harboring CTLA4Ig-IRES 2-Ikappa-Balpha gene in ECV-304 cells].

AIM: To construct adenovirus vector harboring CTLA4Ig-IRES 2-IkappaBalpha gene,and investigate its expression in ECV304 cells. METHODS: Recombinant adenovirus vector harboring CTLA4Ig-IRES 2-IkappaBalpha was constructed by homologous recombination in E.coli BJ5183. Then recombinant vector was packaged and propagated in 293 cells. ECV304 cells were infected with the recombinant adenovirus, and Western Blot was used to detect IkappaBalpha and CTLA4Ig protein expression in infected ECV304 cells. The effect of expressed IkappaBalpha and CTLA4Ig on the expression of TNF-alpha of ECV304 cells stimulated with LPS was also investigated. RESULTS: The recombinant CTLA4Ig-IRES 2-IkappaBalpha adenovirus was generated by homologous recombination and identified by PCR methods. Ikappa-Balpha and CTLA4Ig were expressed in ECV304 cells infected with the recombinant adenovirus. The production of TNF-alpha induced by the LPS was inhibited in infected ECV304 cells. CONCLUSION: The recombinant CTLA4Ig-IRES 2-IkappaBalpha adenovirus may offer a method to down regulate the inflammatory cytokines in the inflammatory reaction which may lay the foundation for genetherapy of immunosuppression.

Abatacept↗

Effect of the interaction between paranoxonase 1 and ATP-binding cassette transporter 1 gene polymorphism on serum lipid level.

OBJECTIVE: To investigate the effect of interaction between paranoxonase 1 (PON1)gene polymorphism and ATP-binding cassette transporter 1 (ABCA1) genetic variation on serum lipid level. METHODS: Polymerase chain reaction-restricted fragments length polymorphism was used to determine PON1 A/B192 and ABCA1R219K genotype of 1019 subjects, including 680 patients with strokes and 339 healthy individuals as controls. RESULTS: No significant association between A/B192 genotype and any of the lipid measurements was detected. The levels of HDL-C in the subjects with RR, RK and KK genotypes showed a significant upward tendency respectively (P < 0.05); the levels of their triglyceride (TG) tended downward respectively, but there were no significant differences between them. The relationship between R219K genotype and serum lipid level was modified by A/B192 genotype. The levels of HDL-C in the subjects with AA/RR genotype and BB/KK genotype [(1.41 +/- 0.40) mmol/L, (1.41 +/- 0.39) mmol/L] were significantly different from that in the subjects with BB/RR genotype [(1.28 +/- 0.36) mmol/L] (P < 0.05). CONCLUSION: The result exhibited an interaction of PON1 A/B192 and ABCA1 R219K on serum lipid level.

ATP-Binding Cassette Transporters↗

[Serum S100 protein and radiation-induced brain injury in astrocytoma patients].

OBJECTIVE: To study the value of serum S100 protein in the diagnosis of radiation-induced brain injury in astrocytoma patients. METHODS: Serum S100 protein levels were detected by enzyme-linked immunosorbent assay in 86 astrocytoma patients in the course of radiotherapy. RESULTS: The positivity rates of serum S100 protein before, in the middle of and after radiotherapy were 0, 9.3% (8/86) and 19.8% (17/86), respectively, showing significant differences (P<0.05) between these time points for measurement. CONCLUSION: High levels of serum S100 protein are associated with radiation-induced brain injury in astrocytoma patients and may serve as the marker for early diagnosis of the injury.

Adult↗

[Establishment of transgenic mouse line skin-specifically expressing hCTLA4-Ig].

hCTLA4-Ig,which is capable of inhibiting T cell activation and rendering T cell to be anergy by blocking co-stimulatory signal pathway, is recognized as a potential therapeutic molecule to extend the survival of skin graft in burn medicine. To investigate whether the survival of skin graft can be further extended by skin-specific expression of hCTLA4-Ig, a skin-specific expression vector was constructed by pacing the encoding sequence of hCTLA4-Ig under the drive of K14 promoter, and a transgenic mouse line was established with the vector. RT-PCR and Northern blot assay indicated that hCTLA4-Ig was skin-specifically expressed in transgenic mice at a rather high level. Compared with the expression of GAPDH, the expression of hCTLA4-Ig remained constant through generations and over lifetime. The results of this paper indicated that a transgenic mouse line skin-specifically and constitutively expressing hCTLA4-Ig had been established.

Abatacept↗

A new phenylethanoid glycoside from Clerodendrum inerme.

A new phenylethanoid glycoside, 2-(3-methoxy-4-hydroxylphenyl) ethyl-O-2",3"-diacetyl-alpha-L-rhamnopyranosyl-(1-->3)-4-O-(E)-feruloyl-beta-D-glucopyranoside, was isolated from the aerial parts of Clerodendrum inerme (L.) Gaertn, together with monomelittoside, melittoside, inerminoside A1, verbascoside, isoverbascoside, campneoside I. Their structures were determined by spectroscopic methods.

Clerodendrum↗

[Estimation of fetal weight on the basis of neural network].

The ultrasonic estimation of fetal weight at delivery is of important prognostic significance in obstetrical practice. The convertional regression formulas used for estimating fetal weight have the disadvantage of less reliability. In this study, we used the back propagation neural network (BP) to estimate Fetal Weight. Some input variables were adopted in constructing the BP model: biparietal diameter (BPD), cerebellum transverse diameter (TCD), abdominal circumference (AC), liver length (LL), femur length (FL), fetal thigh soft tissue thickness (FSTT), and gestational age (GA). The fetal weights of 109 singleton fetuses were estimated. In the training group and validation group, coincidence rates were 89.77% and 76.19% respectively. The results show that the estimation based on neural network is more accurate than that by regression method. GA, its unit is not week but day in our formulas, is very valuable in combination with other ultrasonic parameters on estimation.

Anthropometry↗

[The relationship between ApoE gene polymorphism and lipid parameters].

OBJECTIVE: To investigate the relationship between apolipoprotein E (ApoE) gene polymorphism and serum lipid profile. METHODS: Polymerase chain reaction-restricted fragments length polymorphism (PCR-RFLP) was used to determine ApoE genotype on 1452 subjects including 1101 cases with cardio cerebrovascular disease including 379 cases with cerebral infarction, 313 cases with cerebral hemorrhage, 257 cases with coronary heart disease, and 152 cases with other types and on 351 healthy controls. RESULTS: After adjusting for age, sex and BMI, the subjects with ApoE4 carriers had significantly higher levels of total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) and ApoB than those with ApoE2 carriers and ApoE3/3 (P < 0.05), and higher level of triglyceride(TG) than those with ApoE3/3 (P < 0.05), while the subjects with ApoE2 carriers had significantly higher levels of high density lipoprotein-cholesterol (HDL-C) than those with ApoE4 (P < 0.05). The effects of ApoE polymorphism exhibited similarity in different sex and age of subjects. Linear regression analysis showed that unlike ApoE3/3, the HDL-C level in ApoE2 carriers tend upward with age (beta = 0.178, P = 0.015), significantly higher than ApoE4 carriers and ApoE3/3 in the cohort of 65-74 years (P < 0.05). The level of TC and TG in ApoE4 carriers had a tendency of downward with age (p = -0.179, P = 0.009; beta = -0.147, P = 0.032). CONCLUSION: ApoE gene polymorphism affected profile of blood lipids and the effects were found in different sex and age. The degrees of effects related to ApoE2 carriers and ApoE4 carriers to blood lipid level seemed to be related to age.

Adult↗

[Blockage of allergic rhinitis in mice with recombinant protein of cytotoxic T lymphocyte associated antigen-4 extracellular domain].

OBJECTIVE: To explore a new immunotherapy against allergic rhinitis. METHODS: The recombinant protein of CTLA4 extracellular domain was obtained through construction of CTLA4-yeast expression system. The allergic rhinitis in mice was induced by sensitizing and challenging with ovalbumin (OVA). The allergic rhinitis related symptoms and the morphological changes in nasal mucosa were compared between the allergic rhinitis group and the CTLA4 extracellular domain group treated with CTLA4 extracellular domain before each challenge by ways of intraperitoneal injection. RESULTS: CTLA4 extracellular domain with a molecular weight of 28 000, which was confirmed by Western blot, could be generated through CTLA4-yeast expression system. The purified CTLA4 extracellular domain could inhibit T cells proliferation in mixed lymphocyte reaction with a inhibitory rate of 95.4%. The mice in allergic rhinitis group appeared typical allergic rhinitis symptoms after OVA challenge, such as rhinorrhea and sneeze. Meanwhile the nasal pathological studies showed edema and congestion in mucosa tissue and local influx of inflammatory cells. Whereas in CTLA4 extracellular domain group, the nasal symptoms were rarely observed, and the pathological change in nasal mucosa was significantly abated. CONCLUSIONS: The protein of CTLA4 extracellular domain could prevent the allergic rhinitis in mice. The underlying mechanism of which might be the inhibition of the T cell activation.

Animals↗

[Isolation and characterization of a HCH degradation Sphingomanas sp. stain BHC-A].

An aerobic bacterium was isolated successfully from a long-term contaminated upland field, which was named BHC-A. The bacterium can utilize Hexachlorocycloexane as a sole carbon source and decompose this substance rapidly and completely. According to its physiological & biochemical characters and the homology analysis of its 16S rDNA sequence, this strain was identified as Sphingomonas sp. Stain BHC-A canmineralize not only alpha, gamma and delta-HCH rapidly, but also 5 mg/L beta-HCH in 12h completely. However, Sphingomonas paucimobilis UT26, a gamma-HCH-decomposing bacterium, can only degrade 12.6% of 5 mg/L alpha-HCH, but not beta-HCH and delta-HCH, even for more than 24h in the medium. The seedling of cucumber was selected as the model plant subject to the inhibition of HCH to the growth of cucurbitaceous plants. The results shows that the degradation of HCH isomers by BHC-A in soil is even steady and excellent, that BHC-A can eliminate the contamination of the soil and the harm to the plants.

Biodegradation, Environmental↗

[Z-ajoene causes cell cycle arrest at G2/M and decrease of telomerase activity in HL-60 cells].

OBJECTIVE: To investigate the molecular mechanisms of Z-ajoene mitosis blocking and telomerase inhibitory effects on HL-60 cells. METHODS: Proliferation inhibition of HL-60 cell line was evaluated by MTT assay. Z-ajoene-induced mitotic blocking effect was investigated by flow cytometry. Immunoblotting analysis was used to determine cell cycle regulatory proteins. The telomerase activity of HL-60 cells was detected by TRAP-silver stain assay. Telomerase hTRT and TP1 mRNA level were determined by RT-PCR. RESULTS: Z-ajoene displayed great proliferation inhibiting effect on HL-60 cells. Progressive increase in the percentage of mitotic block at G(2)/M phase was observed from 4 h to 12 h after treatment with 10 micromol/L Z-ajoene, with a peak at 10 h, which was 1.95 times higher than that in control. Z-ajoene also caused an increase in cyclin B1 accumulation and a decrease of p34(cdc2) expression. But Z-ajoene did not change the level of cyclin A. After treating with 10 micromol/L Z-ajoene for 24 h, the telomerase activity of HL-60 cells was also decreased in a dose-independent manner. Furthermore, telomerase hTRT and TP1 mRNA levels decreased after 10 micromol/L Z-ajoene treatment for 24 h. CONCLUSION: The results suggest that Z-ajoene has potent anti-cancer activity, and that its inhibitory effect on telomerase activity and on cell growth might be the result of G(2)/M phase blocking.

Antineoplastic Agents, Phytogenic↗

[Pilot study on the mechanisms of growth inhibitory effect of cinobufagin on HeLa cells].

OBJECTIVE: To study the effect of cinobufagin (CBG) on HeLa cell proliferation, and to analyze its mechanism. METHODS: Proliferation inhibition in vitro was evaluated by MTT and Sulforhodamine B (SRB) assays in several human tumor cell lines, including Bel-7402, HeLa, MCF-7, BGC-823 and HL60. The cycle of HeLa cells was analyzed by flow cytometry. Two-dimensional electrophoresis was applied to analyze the influence of CBG on HeLa cell proteomics. RESULTS: CBG had inhibitory effects on proliferation of five human cancer cell lines, and the IC(50) values were 0.011 micromol/L (Bel-7402), 0.019 micromol/L (HeLa), 0.116 micromol/L (MCF-7), 0.149 micromol/L (BGC-823) and 1.369 micromol/L (HL60), respectively. HeLa and Bel-7402 cells were among the most sensitive. Flow cytometry assay indicated that the treatment of HeLa cells with various concentrations of CBG for 72 h was able to increase the cell number at G(2)/M phase, from 17.3% up to 35.6%. The results of two-dimensional electrophoresis showed that treatment of HeLa cells with 0.02 micromol/L CBG for 48 h resulted in apparent changes of certain small molecular weight (30,000 - 90,000) acidic proteins (pH 4 - 6). CONCLUSION: Cinobufagin has significant inhibitory effect on growth of five human cancer cells in vitro. It may lead to cell cycle arrest of HeLa cells at G(2)/M phase. It can also change the expression of some small molecular acidic proteins in HeLa cells.

Antineoplastic Agents↗

The role of erythropoietin in regulating angiogenesis.

Erythropoietin (EPO) is an essential growth factor that regulates erythrocyte production in mammals. In this study, we demonstrate a novel role of EPO in regulating angiogenesis in vivo. Epo and Epo receptor (EpoR) are expressed in the vasculature during embryogenesis. Deletion of Epo or EpoR leads to angiogenic defects starting at E10.5, 2 days before ventricular hypoplasia and 3 days before the onset of the embryonic lethal phenotype. Overall, angiogenesis was severely affected in the mutant embryos: vascular anomalies included decreased complexity of the vessel networks. However, de novo vasculogenesis remained intact, consistent with the differential expression of Epo and EpoR during the early stages of embryonic development. The aforementioned angiogenesis defect can be partially rescued by expressing human EPO during embryogenesis. Moreover, Ang-1 expression is regulated by EPO/EPOR under normoxic conditions. Taken together, our results suggest important roles of EPO and EPOR in angiogenesis.

Angiopoietin-1↗

Nontranscriptional modulation of intracellular Ca2+ signaling by ligand stimulated thyroid hormone receptor.

Thyroid hormone 3,5,3'-tri-iodothyronine (T3) binds and activates thyroid hormone receptors (TRs). Here, we present evidence for a nontranscriptional regulation of Ca2+ signaling by T3-bound TRs. Treatment of Xenopus thyroid hormone receptor beta subtype A1 (xTRbetaA1) expressing oocytes with T3 for 10 min increased inositol 1,4,5-trisphosphate (IP3)-mediated Ca2+ wave periodicity. Coexpression of TRbetaA1 with retinoid X receptor did not enhance regulation. Deletion of the DNA binding domain and the nuclear localization signal of the TRbetaA1 eliminated transcriptional activity but did not affect the ability to regulate Ca2+ signaling. T3-bound TRbetaA1 regulation of Ca2+ signaling could be inhibited by ruthenium red treatment, suggesting that mitochondrial Ca2+ uptake was required for the mechanism of action. Both xTRbetaA1 and the homologous shortened form of rat TRalpha1 (rTRalphaDeltaF1) localized to the mitochondria and increased O2 consumption, whereas the full-length rat TRalpha1 did neither. Furthermore, only T3-bound xTRbetaA1 and rTRalphaDeltaF1 affected Ca2+ wave activity. We conclude that T3-bound mitochondrial targeted TRs acutely modulate IP3-mediated Ca2+ signaling by increasing mitochondrial metabolism independently of transcriptional activity.

Animals↗

Use of ferrocenyl surfactants of varying chain lengths to study electron transfer reactions in native montmorillonite clay.

A series of ferrocenyl surfactants was tested as model compounds to study electron transfer reactions involving structural Fe(III) in clay minerals. The surfactants contain trimethylammonium headgroups, ferrocene tail groups, and intervening hydrocarbon chain lengths of one, six, or 11 carbons. Two factors considered to be decisive for electron transfer were addressed: (1) physical access of the surfactant ferrocene to the reactive sites through hexagonal holes in the clay lattice by X-ray diffraction (XRD) and small-angle X-ray scattering (SAXS) and (2) thermodynamic favorability of the overall oxidation/reduction reaction based on experimentally determined oxidation/reduction potentials. In suspensions of clay with the longer chain surfactants, (ferrocenylhexyl)trimethylammonim (FHTMA+) and (ferrocenylundecyl)trimethylammonium (FUTMA+), where electron transfer may be expected to be favored by both factors, physical accessibility, and thermodynamic favorability, ferroecene oxidation was observed by diffuse reflectance infrared spectroscopy (DRIFT), ultraviolet-visible spectroscopy (UV-vis), and visual color changes. In contrast, the shorter chain length surfactant, (ferrocenylmethyl)trimethylammonium (FMTMA+), did not participate in electron transfer with the clay, as substantiated by UV-vis and no visible color changes. Rigid conformation and/or higher oxidation/reduction potential than clay Fe can accountforthe lack of reaction. The utility and limitations of using these surfactants as model compounds is discussed.

Aluminum Silicates↗