Objective To investigate whether the release of fibroblast growth factor-1 ( FGF-1 ) was changed after inhibition of S100A13 gene (small hairpin RNA, shRNA)and serum-deprivation in human thyroid cancer cells (TT cells ). Methods The S100A13-shRNA pENTRTM/U6 entry vector was transfected into TT cells. The expression of S100A13 mRNA and protein was detected by immunoflurescence, real-time RT-PCR, and Western blot. Then TT cells were treated with S100A13 gene inhibition and serum-deprivation. The changes in release of FGF-1 were detected by indirect immunoflurescence, RT-PCR, and ELISA. Results S100A13 shRNA transfected TT cells (S100A13 RNAi cells)had a reduction of S100A13 gene and protein expression by 80%.Indirect immunofluorescence indicated FGF-1 was mostly localized in the cytoplasm and nucleus of TT cells in primary culture. When serum-deprivation stress was given to TT cells, FGF-1 in cytoplasm almost disappeared in the cells at 6 h. RT-PCR indicated that when serum-deprivation stress was given to TT cells the mRNA of FGF-1 was reduced. ELISA showed that with inhibition of S100A13, the release of FGF-1 was reduced (P＜0.05).Conclusion S100A13-shRNA pENTRTM/U6 entry vector transfected TT cells may inhibit the expression of S100A13 and reduce the release of FGF-1.

Objective To explore the alterations of protein phosphatase-2A (PP-2A) in lymphocytes in mild cognition impairment (MCI) and Alzheimer's disease (AD).Methods The activity PP-2A of was measured by ~(32)p liquid seintillography for incorporated radioactivity in control group(n=11) , the MCI group(n=11),and the AD group(n=11).The expression of PP-2A was determined by Western blot.Results In the control group,the activity of PP-2A (1.01?0.09) and the expression of PP-2A (0.96?0.07) were high while in the MCI group,the activity of PP-2A (0.71?0.12) and the expression of PP-2A (0.80?0.05) were decreased (both P

Objective To construct and express the recombinant plasmid pET32α-Sj26GST of Schistosoma japonicum(sj)in Escherichia coli(E.coli)B121(DE3).Methods The total RNA was extracted from sj adult worms by ultrasound-breaking,Sj26GST antigen gene was amplified by RT-PCR from the total RNA,then cloned into prokaryotic expression plasmid pET32α(+) and transformed into E.coli B12(DE3)to construct pET32α-Sj26GST;BL21(pET32α-Sj26GST)WaS induced with isopropyl-β-D-thiogalactopyranosid(IPTG),and the expressed products were analyzed and identified by SDS-PAGE and Western blot.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and cloned into pET32α(+)by restriction analysis and PCR identification,the recombinant plasmid pET32α-Sj26GST was successfully constructed;the relative molecular mass of the expressed recombinant protein was approximately 49×103 by SDS-PAGE,and the amount of the expressed protein was 24%of the total bacterial proteins;the fusion protein could be recognized by sera from rabbits infected with sj by Western blot.Conclusions The recombinant plasmid pET32α-Sj26GST is successfully constructed and highly expressed in E.coli in fused form with Trx-tag and His-tag,and the expressed fusion protein shows specific antigenicity.

OBJECTIVETo evaluate the effect of the shade and thickenss of luting agent on the final color of different thickness Solidex resin in vitro.

METHODSFive specimens of Solidex resin were fabricated at thickness of 0.5, 1.0 and 1.5 mm. The pieces of Ni-Cr alloy, two shade composite resin and opaque porcelain veneered Ni-Cr alloy were used as backgrounds. The CIE L*a*b* values of specimens were measured by a colorimeter (ShadeEye NCC) to determine the colorimetric difference, when different thickness of specimens were overlaid the different shade and thickness of luting agent at the same backgrounds.

RESULTSThe final color difference of specimens was significantly influenced by the factors of luting agent and the interaction of specimens thickness, backgrounds and luting agent(P< 0.001). Regarding the thickness of luting agent, A3 shade luting agents produced clinically unacceptable difference (deltaE*>2) in the final color of Solidex specimens at the thickness of 0.5 mm. Regarding the shade of luting agent, the color difference was more than 2 when the Solidex specimens at the thickness of 0.5 mm or 1.0 mm were against the 0.2 mm thick luting agent.

CONCLUSIONThe final color of thinner Solidex specimens was significant affected by the change of the shade and thick of luting agent.

OBJECTIVETo analyze the final color of Solidex in six thickness interacting with three different backgrounds.

METHODSFive specimens of Solidex in each shade A1, A3, and C2 were fabricated at thickness of 1.0, 1.2, 1.4, 1.6, 1.8, and 2.0 mm. The CIE L(*)a(*)b(*) values of specimens were measured using a colorimeter (ShadeEye NCC) to determine the colorimetric difference, when the specimens overlaid on different backgrounds.

RESULTSWhen the thickness was more than 1.8 mm, the color difference (DeltaE(*)ab < 1) was not perceived by human observers. When the thickness was 1.4 mm or less than 1.4 mm, the color difference (DeltaE(*)ab > 2.72) was considered clinically unacceptable between alloy background and two types of composite resin background.

CONCLUSIONSAlloy background had more evident influence on the final color of Solidex than composite resin background. When making metal-free crown restoration using Solidex, it was advisable to make the composite resin more than 1.8 mm thick.

Adult ; Brain Diseases ; chemically induced ; Ethylene Dichlorides ; poisoning ; Female ; Humans ; Male ; Middle Aged ; Occupational Diseases ; chemically induced

Objective To analyze the pathological and clinical characteristics of patients with idiopathic IgA nephropathy accompanied by vasculitic/crescentic lesion (IgA-V/C). Methods Data of 222 patients diagnosed as idiopathic IgA nephropathy by renal biopsy, among them 87 cases with vasculitic/crescenlic (V/C)lesion, from our department in 2004 were analyzed retrospectively. Clinical and pathological data from patients with IgA-V/C were compared to those of non-IgA-V/C patients and of lupus nephritis (LN) patients with V/C lesion. Results Vasculitic/crescentic lesion was found in 39.19% (87/222) patients with idiopathic IgA nephropathy.And about( 14.08?12.75)% of the glomeruli was affected. It should be taken into account that there was no significant differences of clinical manifestations including blood pressure, urinary protein excretion between IgA-V/C group and non-IgA-V/C group .except serum creatinine(Scr)level which was significantly higher in IgA-V/C group. In addition, only 37.9% of IgA-V/C patients presented high Scr level,thus the lesion of V/C in idiopathic IgA nephropathy was easily overlooked. Patients with idiopathic IgA nephropathy were found to have higher percentage of glomerular sclerosis (64.86% vs 40.00%) and ratio of sclerostic glomeruli to total glomeruli [ (26.98 ?24.68)% vs (16.10 ?18.80)% ]as compared to LN group, which further predicated the progressing characteristics of IgA nephropathy. Conclusions Vasculitic/crescentic lesion is a quite common finding in idiopathic IgA nephropathy and often associated with no dramatically symptoms. It is possible for vasculitic/crescentic lesion to induce unmarked lose of nephron slowly and continually, so as to accelerate IgA nephropathy progression to end-stage renal failure.

AIM: To study apoptosis and bcl-2 mRNA gene expression of cardiomyocytes in donor hearts of immature rabbits underwent prolonged protection by 11, 12-epoxyeicosatrienoic acid (11, 12-EET), and further probe into the possible mechanisms. METHODS: 24 isolated immature rabbit hearts were performed to the model in a Langendorff perfusion apparatus and randomly assigned to normal control group,ST control group and EET group. The isolated rabbit hearts in ST control group and EET group were stored for 24 hours with 4 ℃ hypothermia, and underwent 30 minutes of reperfusion (37 ℃). TUNEL and in situ hybridization (ISH) methods were applied in the present study and apoptotic cells and bcl-2 mRNA gene expression were observed. RESULTS: The numbers of apoptotic cardiomyocytes in ST group and EET group were higher than that in normal control group, and the numbers of apoptotic cardiomyocytes were significantly decreased in EET group and bcl-2 mRNA positive expression were higher than that in ST control group, respectively. CONCLUSIONS: There were apoptosis during the prolonged protection of donor heart in our study, and we proved that: ①11,12-EET could decrease cardiomyocyte apoptosis significantly. ②Up-regulation of the bcl-2 mRNA expression in cardiomyocytes may be one of the mechanism responsible for inhibition of cardiomyocyte apoptosis by 11, 12-EET.

Objective To construct and express the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum(Sj) in Escherichia coli BL21 (DE3). Methods Total RNA was extracted from Sj adult worms by ultrasound-breaking, Sj26GST and Sj32 antigen gene was respectively amplified by RT-PCR from the total RNA; Sj26GST-Sj32 fusion gene obtained with gene splicing by overlap extension(SOEing) was cloned into prokaryotic expression plasmid pET28α and transformed into Escherichia coli BL2 (DE3) to construct pET28α-Sj26GST-Sj32;BL21 (pET28α-Sj26GST-Sj32) was induced with isopropyl-β-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)and Western blotting. Results The 1991 bp Sj26GST-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into pET28α by restriction analysis and PCR identification, the recombinant plasmid pET28α-Sj26GST-Sj32 was successfully constructed; the relative molecular mass of the expressed recombinant protein was approximately 69 × 103 by SDS-PAGE, and the amount of the expressed protein was 25% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pET28α-Sj26GST-Sj32 is successfully constructed and highly expressed in Escherichia coli in fused form with His-tag, and the expressed fusion protein shows specific antigenicity.

An investigation on the chemical constituents of the 90% EtOH extract of Perovskia atriplicifolia led to the isolation of fifteen compounds from the EtOAc fraction. Based on the detailed spectral analysis (MS, 1D and 2D NMR), as well as comparison with the literatures, the structures of compounds 1-15 were determined as cirsimaritin (1), salvigenin (2), syringaldehyde (3), vinyl caffeate (4), 2α, 3α-dihydroxyolean-12-en-28-oicacid (5), 2α, 3α-dihydroxyurs-12-en-28-oicacid (6), niga-ichigoside F1 (2α, 3β, 19α, 23- tetrahydroxyurs - 12-en-28-oicacid- O-β-D- glucopyranoside, 7), sericoside (8), 4-epi-niga-ichigoside F1 (2α, 3β, 19α, 24-tetrahydroxyurs-12-en-28-oicacid O-β-D-glucopyranoside, 9), 2α, 3β, 24-trihydroxyolean-12-en-28-oicacid O-β-D-glucopyranosyl-(1 --> 2) - β-D-glucopyranoside (10), pruvuloside A (11), asteryunnanoside A [2α, 3β, 23-trihydroxyolean-12-en-28-oicacid O-β-D-glucopyranosyl-(1 --> 2)-β- D- glucopyranoside,12], rosmarinic acid methyl ester (13), β-sitosterol (14), and daucosterol (15), respectively. Compounds 1-13 were isolated from the Perovskia genus for the first time. All the compounds were obtained from P. atriplicifolia for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization