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Yttrium vanadate:europium nanoprobes (YVO4∶Eu NPs) with good fluorescence properties and water solubility were synthesized by solvent thermal method.Due to the overlapping of the excitation spectrum of YVO4∶Eu NPs and the absorption spectrum of tryptophan, fluorescent internal filter effect (IFE) occurred, in which YVO4∶Eu NPs were the fluorophore and tryptophan was the absorber, leading the fluorescence of YVO4∶Eu NPs was quenched.Therefore, a new method for the determination of tryptophan was established by using fluorescent YVO4∶Eu as nanoprobes based on IFE.Some experimental parameters, such as the adding amount of YVO4∶Eu NPs, pH value of the reacting solution, and reacting time, were investigated.Under the optimum reaction conditions, the linear range of the method was 4.0×10-6-4.0×104 mol/L and the detection limit was 1.0×10-6 mol/L (3σ).The content of tryptophan in the soy sauce was determined with the recovery of 95.2% and 97.3%.This method is simple, rapid, sensitive and accurate.
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Objective To investigate the value of serum thyroglobulin (Tg) and antithyroglobulin antibody (TgAb) in differentiated thyroid carcinoma complicated with Hashimoto's thyroiditis after thyroid ablation.Methods Serum Tg and TgAb levels and the status of illness in 154 differentiated thyroid carcinoma patients with coexistent Hashimoto's thyroiditis and confirmed pathology after surgery followed by remnant ablation were performed during three years follow up.Tg and TgAb levels were assessed by chemiluminescent immunoassay assay.The cases were divided into three groups (according to the level of Tg):Tg ≤ 1 μg/L group,1 μg/L＜Tg ≤ 10 μg/L group and 10 μg/L＜Tg≤ 100 μg/L group.TgAb＞40 kIU/L was considered as positive,Cox's proportional hazard model was used to analyse prognostic value in different levels of Tg and TgAb for disease-free survival and recurrence.Results Compared with 1 μg/L＜Tg≤ 10 μg/L group and 10 μg/L＜Tg≤ 100 μg/L group,the relative risk in reflecting cancer recurrence (TgAb＞40 kIU/L) in Tg ≤ 1 μg/L group was 27.000 (95 ％ CI 6.727-108.374).The value of TgAb＞40 kIU/L in Tg≤ 1 μg/L group was greatly increased and highly correlated with metastasis.However,In the condition of Tg＞ 1 μg/L,the disease will be based on the level of TgAb.Conclusion The value of TgAb＞40 kIU/L in Tg ≤ 1 μg/L group seems to be the optimal cutoff value correlated with recurrence and metastasis of differentiated thyroid carcinoma.
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Objective To investigate the patterns of change in thyroid functional parameters ( serum TSH,FT3, and FT4 ) in patients with papillary thyroid cancer (PTC) before and after the initial 131I treatment for thyroidal remnant ablation. Methods Seventy-four PTC patients, treated with 3.7 GBq 131 I therapy, were divided into two groups, group A with serum TSH＜30 mIU/L and group B with serum TSH ≥30 mIU/L the day before 131I treatment. Five days after the treatment, the patients were re-examined for serum FT3, FT4, and TSH levels.Results In group A (22 cases), 5 days after the 131I ablation treatment, FT4significantly increased by 88% and FT3 by 87%, while TSH decreased by 87% (all P＜0. 05 ), and 45% (10/22)cases manifested the signs of transient thyrotoxicosis. In group B (52 cases)after treatment, individual variance of FT3 and FT4 was obvious,with FT4 decreased by 13% and FT3 decreased by 14% ( both P＜0. 05 ), while TSH slightly increased by an average of 6% ( P＞0.05 ). Conclusion After the initial 131 I ablation therapy for thyroidal remnant, the thyroid hormone levels in some PTC patients significantly increase while in others may slightly decrease in the early stage. The supplementary and suppressive therapy after 131I ablation for PTC patients might be individualized depending on the thyroid hormone determination.
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Objective:To study the differences of bleomycin-induced pulmonary fibrosis in mice induced by intraperitoneal injection and intratracheal instillation of bleomycin.Methods:ICR mice (male,8 w of age,18 to 22 g bodyweight) were used.①ICR mice were randomly divided into three groups.In the group P,bleomycin was injected intraperitoneal five times in a dose of 40 mg/kg,and in the group I,bleomycin was instilled intratracheally in a dose of 5 mg/kg.The mice were killed at 14,28 or 40 day.②ICR mice were randomly divided into four groups : bleomycin was injected intraperitoneal three,four or five times in a dose of 40 mg/kg,and in the group control ,bleomycin was injected intraperitoneal in a dose of 200 μl for five times.The mice was killed at 28 or 40 day.The pathological changes and symptoms were observed.Results:① The weight of the mice given blemycine were lost after injection.Symptoms were more serious in group P than those of group I.The pulmonary coefficient of group P was more than that of group I.At 28 day after injection,fibrosis was widely and stably formed mainly in around the bronchia and bronchioles,especially,near the pulmonary hilar area,in the group I mice,however,the same changes were mainly seen under the pleura or perivascular pulmonary tissues in the mice of group P.The pathological score of pulmonary fibrosis was different between those two groups.②Different dose of bleomycine induced different change of the mouse's weight.The most of all of the three group were five time injection; The lung index of five time injection of bleomycine was the most.The pulmonary fibrosis mouse model was made successfully.After comparising all of the data,we found intraperitoneal of bleomycine five times was the more convenient method.Conclusion:The sites of pulmonary fibrosis in the mice are different between the mice induced by intraperitoneal injection or intratracheal instillation of bleomycin.Intraperitoneal injection five times is a more convenient method to make pulmonary fibrosis.
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Objective:To study the clinical application potential of aFGF and try to produce aFGF as gene engineering medicine.Methods:The complete encoding cDNA of human aFGF was isolated from human lung fibroblast with RT PCR.Recombinant human aFGF was expressed in secretory pichia expression system and purified with heparin sepharose chromatography.The activity of aFGF was detected in NIH3T3 proliferation assay,chick embryo chorioallantoic membrane assay(CAM) and wound healing assay.Results:The recombinant aFGF was expressed in large quantity(yield=12 mg/L) and was capable to promote proliferation of NIH3T3,angiogenesis and wound healing.Furthermore,those activities of aFGF could be agonized by recombinant FGFR extracellular domain.Conclusion:Recombinant human aFGF was expressed efficiently and possessed natural biological activities.
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Objective:To clone and express mouse fibroblast growth factor eight(FGF 8) and investigate the function of it Methods:According to the published sequence of mouse fibroblast growth factor 8, a pair of special primers was designed Then the full length cDNA of FGF 8b (a predominant isoform of FGF 8) was isolated from the total RNA of the mouse embryo by means of reverse transcription PCR and subcloned into the yeast expression vector of pYEX4T 1 in correct direction It was identified with sequencing The recombinant plasmid was transformed into yeasts The fusion protein expressed was verified by using Western blot The activity of the protein was examined by MTT and 3H TdR incorporation Results:The cDNA fragment was about 600 bps, and proved to be "b" isoform of FGF 8 exactly This fusion protein, with molecular weight about 55 kD, was highly expressed in yeast system The data of MTT and 3H TdR incorporation in NIH3T3 cells were increased obviously by the protein, and decreased by the antagonist Conclusion:Fibroblast growth factor eight was cloned and expressed.The protein was found to be capable of stimulating the proliferation of NIH3T3 cells Moreover, the recombinant protein of the extracellular fragment of FGFR1 can antagonize the response of NIH3T3 to the recombinant protein
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Objective: To express recombinant human angiostatin for further application in clinic. Methods: The complete encoding eDNA of human angiostatin was isolated from human embryo liver with RT-PCR and expressed in secretory Pichia expression system. Recombinant human angiostatin was purified with heparin sepharose chromatography and its activity was determined in chick embryo chorioallantoic membrane (CAM) and wound healing assays. Results: Expressed in large quantity (yield=5 mg/L) and purified with heparin sepharose, recombinant angiostatin was showed to have a molecular weight of 43 kD in SDS-PAGE and potently inhibit angiogenesis and wound healing. Conclusion: Recombinant human angiostatin was expressed efficiently in a biologically active form.
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Objective To find the changes of human fibroblast growth factor receptor 1 genone during development.Method Southern blot analysis of genomic DNA isolated from adult and fetal tissues. Result Adult FGFR1 gene structure is different from its embryonic counterpart. Conclusion The disserence might lead to changes of FGFR1 expression as wel as functions of the cells.
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Abstract Objective:To construct bmcella melitensis Br.melitensis genomic DNA expression library and study its immune effect.Meth-ods: Hind Ⅲ digested Br.melitensis genomic DNA fragments were cloned into pSV-?-Gal plasmds.The recombinant plasmids were transformedinto JM109 host bacteria. After large-scale isolation, the recombinant plasmids were inoculated into the quadriceps muscle of mice. In the 9thday after third boost immunization .agglutination antibodies against Br. melitensis and lymphocyte mitogenic ability of mice were determined. Re-sults : Expression library nucleic acid vaccine can induce the production of agglutination antibodies against Br. melitensis and stimulate the lym-phocyte porliferation primed by ConA. Conclusion:The genomic DNA expression library may provide a new vaccine against Br. melitensis infec-tion.
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Objective: To express recombinant human soluble fibroblast growth factor receptor 1 ( sFGFR1) and study its antagonistic activity on FGF. Methods: Human sFGFR1 cDNA, isolated from human lung fibroblast cells with RT-PCR was confirmed by DNA sequencing and cloned into pYEX4T-1 yeast expression vector. The recombinant sFGFR1 was expressed in DY150 yeast cells and the product was identified by SDS-PAGE and Western blot. The activity of recombinant sFGFR1 was detected in N1H3T3 proliferation inhibition assay. Results: GSF-sFGFR1 fusion protein was expressed in yeast cells and was observed as a band of 60 Id) on a SDS- PAGE gel and by Western blot. The recombinant fusion protein was also found to be able to suppress FGF-induced proliferation of NIH3Th cells. Conclusion: Recombinant human GST-sFGFR1 fusion protein was expressed in yeast efficiently and showed natural biological activities.
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Using human acidic fibroblast growth factor Specific primers we tested the technique of screening cDNA library by a DNA polymerase chain reaction.With the method,we have suc-cessfully isolated encoding region cDNA of human interleukin 8 from both human dermal fibrob-last and PMA stimulated U937 cell cDNA libraries.
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TNF? mRNA in human bronchoalveolar lavage(BAL)cells was quantitated by a olymerase chain reaction(PCR)using a cRNA internal standard,cRNA molecules were in vitro transcribed from pAW108 plasmid in which the sequences of upstream primer and omolementary sequences of downstream primer of TNF? were inserted.The cRNA and mRNA extracted from BAL cells were mixed and reverse-transcribed into cDNA.The resultant cDNA mixture was 1:3 diluted and amplified by PCR using TNF? specific primers.~(32)p-labeled upstream primers were included in the PCR reaction,cDNA fragments amplified was run on a 3% agarose gel.The radioactivity of positive bands was determined in a scintillation ounter.After plotting variable template concentrations of the internal standard pAW108 cRNA and the number of BAL cells against the radioactivity of their PCR products,the levels of TNF? mRNA in BAL cells were quantitated by comparision to those of cRNA internal standard.
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Objective:To investigate the mechanisms of Mitoxantrone(MTN) induced apoptosis in human acute myloid leukemia HL 60 cells.Methods:Exposured log phase growth HL 60 cells to different Mitoxantrone concentrations for different time respectively and then test the inhibitive effect by modified tertrozalium salt(MTT) assay.Morphologic evidence for apoptosis of MTN induced on HL 60 cells was determined by transmission electron microscope.The detection of MTN induced internucleosamal DNA fragmentation by agurose gele lectrophoresis.The effects on HL 60 cells proteins(Bcl 2,Bax) implicated in apoptosis were determined by Flow cytoetric.Results:(1)The antiproliferative effects were observed following exposure micromilligram level of MTN.It shows a very substantial differences in the dose and duration dependency of the observed antiproliferative and cytotoxic effects.(2)The HL 60 cells treated by MTN are observed apoptosis characteristic morphological changes such as complete cell membrane,nuclear fragmention apoptosic bodies and found about 200 bp trapezoidal belt with DNA electrophoresis.(3)The protein test results of Bcl 2 and Bax indicate that when the time of MTN treating HL 60 cells is the same,Bcl 2 protein and the ratio of Bcl 2 protein and Bax protein(Bcl 2/Bax) descend along with the ascending concentration of MTN.Bax protein has no connection with the concentration of MTN.When the concentration of MTN treating HL 60 cells are the same.The ratio of Bcl 2/Bax descends as the time of MTN treating HL 60 cells is prolonged.Conclusion:MTN has the function of inducing apoptosis on HL 60 cells with descent of Bcl 2 and Bcl 2/Bax.The regulation pathway of Bcl 2 may be a mechanism of MTN induced apoptosis on HL 60 cells.