HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.

Objective: To optimize the preparation technology of paeoniflorin microcapsule (PM), and to study its behavior of in vitro release. Methods: Using encapsulation efficiency and drug loading as indicators, the PM were prepared by complex agglutination method, the preparation technology of PM was optimized by Doehlert design. The dissolution volume of PM within 20 h and the release curve were measured by HPLC, afterwards its morphology and particle size were studied by electron microscopy. Results: The encapsulation efficiency and drug loading were significantly related to the ratio of coating material, stirring speed, and pH value. The optimal conditions were as follows: the ratio of coating material to paeoniflorin was 4.3:1, the stirring speed was 305 r/min, and the pH value was 4.0. The obtained microcapsules were smooth round capsule-shaped, with non-adhesions and uniform particle size, the encapsulation efficiency was up to 83.81%, the drug loading was 24.24%, and the microcapsule diameter was below 200 μm, with sustained-release effect. Conclusion: The complex agglutination method is simple and reliable to prepare PM, and the product is stable. As a new formulation, microcapsule has a broad prospect for development.

Emerging data indicated that HCV subverts the antiviral activity of interferon (IF); however,whether HCV core protein contributes to the process remains controversial. In the present study, we examined the effect of HCV core protein on interferon-induced antiviral gene expression and whether the effect is involved in the activation and negative regulation of the Jak/STAT signaling pathway. Our results showed that, following treatment with IFN-α, the transcription of PKR, MxA and 2'-5'OAS were down-regulated in HepG2 cells expressing the core protein. In the presence of HCV core protein,ISRE-dependent luciferase activity also decreased. Further study indicated that the core protein could inhibit the tyrosine phosphorylation of STAT1, whereas the level of STAT1 expression was unchanged.Accordingly, SOCS3, the negative regulator of the Jak/STAT pathway, was induced by HCV core protein. These results suggests that HCV core protein may interfere with the expression of some interferon-induced antiviral genes by inhibiting STAT1 phosphorylation and induction of SOCS3.

This paper reported that the activation of oncogenes in human fetal esopha geal epithelium treated by alternariol (AOH). It was found that NIH/3T3 cells were transformed via transfeetion of DNA extracted from human fetal esophageal epithelium which was cultured and treated by 10?g/ml AOH in a short term in vitro. The efficiency of primary loci was 0.17 focus per ?g of DNA. In the secondary transfection, the efficiency was 0.58 focus per ?g of DNA (P

Objective The 4-and 16-hydroxylated metabolites of estrogens have been implicated in carcinogenesis,whereas its 2-hydroxylated metabolites have been shown to have antiangiogenic effects.We aimed to examine whether the polymorphisms of catechol-O-methyltransferase(COMT)involved in the estrogen metabolism are associated with endometrial cancer risk.Methods Polymerase chain reaction- restrictive fragment length polymorphism(PCR-RFLP)analysis was used to study the variant allele frequency distributions of COMT Val158Met genetic polymorphism in a population based case-control study with 132 endometrial cancer cases and 110 controls.Odds ratios(OR)and 95% confidence intervals(CI) were estimated by unconditional logistic regression after adjustment for known or suspected risk factors for endometrial cancer.Results The most frequent genotype was COMT~(Val/Val)(47.2%,52/110)in control group and COMT~(Mal/Met)(58.3%,77/132)in endometrial cancer group.The difference between the two groups was of statistical significance(P

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo amplify the full-length cDNA and characterize the structure and biological function of a novel expression sequence tag ST55 (GenBank Accession No. BM121646).

METHODSRapid amplification of cDNA ends was used to clone the full-length of cDNA of ST55 in this study. Then, its tissue distribution was checked by Northern blots, and the associated protein was screened by GAL 4-based yeast two-hybrid. The effect of stable transfection of the cDNA on cell proliferation was evaluated in ECV304 cells.

RESULTSA full-length 1404 bp cDNA was cloned, and it was accepted as a novel human mRNA by GenBank (No. AY074889), named endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1). Bioinformatic analysis found that the EOLA1 encoded 158 amino acids, 17.89 kDa protein, and mapped to chromosome Xq27.4 with 5 exons. EOLA1 expressed in different human normal tissues and cancer cell lines. Using the EOLA1 cDNA as bait, we performed a yeast two-hybrid screening of a human liver cDNA library and identified metallothionein 2A (MT2A) as associated protein. The interaction between EOLA1 and MT2A was confirmed by co-immunoprecipitation experiments. Stable transfection of EOLA1 was noted to stimulate ECV304 cell proliferation (P < 0.05).

CONCLUSIONThe findings suggest that EOLA1 is a novel gene and the interaction of EOLA1 and MT2A may play an important role in cell protection in inflammation reaction.

Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Blotting, Western ; Cell Line ; Cell Proliferation ; Chromosomes, Human, X ; genetics ; Exons ; genetics ; Humans ; Immunoprecipitation ; Membrane Proteins ; genetics ; metabolism ; physiology ; Metallothionein ; genetics ; metabolism ; Molecular Sequence Data ; Protein Binding ; Sequence Alignment ; Two-Hybrid System Techniques

OBJECTIVETo investigate the effect of pH value and fluoride ions on the corrosion resistance of pure Ti and Ni-Cr-Ti alloy in the artificial saliva.

METHODSElectrochemical technique was used to measure the electric potential of corrosion (Ecorr), current density of corrosion (Icorr) and polarization resistance (Rp) of pure titanium and Ti-Ni-Cr alloy in the artificial saliva with different pH value and fluoride concentrations. After electrochemical analysis, microstructure and phase diffraction were examined by FSEM.

RESULTSWith the lower pH value, the Ecorr and Icorr of pure titanium and Ti-Ni-Cr alloy increased, the Rp decreased, there was a significant difference (P<0.05). The Ecorr and Icorr increased markedly, the Rp significantly reduced in the artificial saliva containing 0.2% NaF (P<0.01). FSEM showed that pure titanium and Ti-Ni-Cr alloy surface corrosion, pure titanium in the artificial saliva containing 0.2% NaF was most serious.

CONCLUSIONLower pH value decreases the corrosion resistance of pure titanium and Ti-Ni-Cr alloy and the artificial saliva containing fluoride ions decreases the corrosion resistance of pure titanium.

Chromium Alloys ; chemistry ; Corrosion ; Dental Alloys ; chemistry ; Electrochemistry ; Fluorides ; chemistry ; Hydrogen-Ion Concentration ; Materials Testing ; Metal Ceramic Alloys ; chemistry ; Nickel ; chemistry ; Saliva, Artificial ; chemistry ; Surface Properties ; Titanium ; chemistry

BACKGROUNDThe anterior cruciate ligament (ACL) is one of the most commonly injured knee ligaments. Even following ACL reconstruction, significant articular cartilage degeneration can be observed and most patients suffer from premature osteoarthritis. Articular cartilage degeneration and osteoarthritis development after ACL injury are regarded as progressive process that are affected by cyclic loading during frequently performed low-intensity daily activities. The purpose of this study was to perform a meta analysis on studies assessing the effects of ACL reconstruction on kinematics, kinetics and proprioception of knee during level walking.

METHODSThis meta analysis was conducted according to the methodological guidelines outlined by the Cochrane Collaboration. An electronic search of the literature was performed and all trials published between January 1966 and July 2010 comparing gait and proprioception of a reconstructed-ACL group with an intact-ACL group were pooled for this review. Thirteen studies were included in the final meta analysis.

RESULTSThere was no significant difference in step length, walking speed, maximum knee flexion angle during loading response, joint position sense and threshold to detect passive motion between the reconstructed-ACL group and the intact-ACL group (P > 0.05). However, there was a significant difference in peak knee flexion angle, maximum angular knee flexion excursion during stance, peak knee flexion moment during walking and maximum external tibial rotation angle throughout the gait cycle between the reconstructed-ACL group and the intact-ACL group (P < 0.05).

CONCLUSIONSStep length, walking speed, maximum knee flexion angle during loading response, joint position sense and threshold to detect passive motion usually observed with ACL deficiency were restored after the ACL reconstruction and rehabilitation, but no significant improvements were observed for peak knee flexion angle, maximum angular knee flexion excursion during stance, peak knee flexion moment during walking and maximum external tibial rotation angle throughout the gait cycle.

Anterior Cruciate Ligament ; surgery ; Biomechanical Phenomena ; Humans ; Knee Joint ; surgery ; Reconstructive Surgical Procedures ; methods ; Walking ; physiology

OBJECTIVETo study the subcellular localization of human endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) protein in endothelial cells.

METHODSHuman umbilical vein endothelial cell strain ECV304 were cultured in vitro. The fusion protein of enhanced green fluorescent protein (EGFP)-EOLA1 expressing plasmid was constructed. Empty plasmid with EGFP at N side (pEGFP-N2) and fusion protein expressing plasmid EGFP-EOLA1 was respectively transfected into ECV304 cells with liposome. After being cultured for 48 hours, the expression levels of EGFP and fusion protein EGFP-EOLA1 in cells were detected with Western blot. The subcellular localization of EOLA1 protein was detected by laser scanning confocal microscope and immunoelectron microscopy.

RESULTSThe EGFP-EOLA1 coexpression plasmid was verified to be successfully constructed by enzyme cutting and gene sequencing. The fusion protein of EGFP-EOLA1 was observed to express in transfected cells through Western blot. Green fluorescence scattered all over the ECV304 cells transfected with empty plasmid and cells transfected with fusion protein expressing plasmid, and it gathered obviously in the nuclei in the latter cells. Immune deposits were observed in the matrix of cells transfected with fusion protein expressing plasmid but not in the cells transfected with empty plasmid.

CONCLUSIONSEOLA1 protein is localized in the nucleus and the matrix of ECV304 cell, and it plays its role as a signal transduction factor.

Cell Line ; Cell Nucleus ; metabolism ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lipopolysaccharides ; metabolism ; Membrane Proteins ; metabolism ; Signal Transduction