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.

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

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.

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

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 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 establish quality standard of Zhuang medicine Lonicera dasystyla, and provide scientific basis for the quality control of L. dasystyla.

METHODCharacteristics of materia medica, microscopic features, TLC indentification, inspection, extractum and determination of chlorogenic acid, macranthoidin B, dipsacoside B were carried out through the experience, microscopic, physical and chemical methods, respectively. The standard of quality control was formulated thereafter.

RESULTThe characteristics of materia medica, microscopic features, TLC indentification were specified, the average contents of water, total ash, acid-insoluble ash, alcohol-soluble extracts, chlorogenic acid were 11.6%, 6.6%, 0.2% , 24.4%, 1.16%, respectively, the total amount of macranthoidin B and dipsacoside B was 3.13%. Quality standard of L. dasystyla was proposed according to experimental results.

CONCLUSIONThe quality of L. dasystyla can be controlled effectively with the quality standard.

Chlorogenic Acid ; analysis ; isolation & purification ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Lonicera ; chemistry ; Oleanolic Acid ; analogs & derivatives ; analysis ; isolation & purification ; Quality Control ; Saponins ; analysis ; isolation & purification ; Solubility

OBJECTIVETo explore the pharmacodynamic and pathological mechanism of eucommia ulmoides oliv in improving erectile function.

METHODSThirty male diabetic rats were randomly divided into three groups: group A (n = 10, escipient group), group B (n = 10, sildenafil group), group C (n = 10, eucommia ulmoides oliv group) and group D (n = 10, the normal control group). After gavage for four weeks, the catching behaviors of all rats were observed, and ultrastructure of myelinated nerve fibers in penile tissue was examined by transmission electron microscope. The expression of neuronal nitric oxide synthase (nNOS) in penile tissues was examined by two steps immunohistochemistry method.

RESULTSCompared with group A, catching frequency of the rats in group C was notably increased (P < 0.05) and the expression of nNOS in penile tissue was significantly (P < 0.001). The examination by transmission electron microscope showed that in the rats' penile tissue of group A, myelinated nerve fibers were irregularly arranged and partially degenerated, and myelin sheaths lamella were splited and exhibited vacuoles or network forms. In group C, there were regular arrangements of myelinated nerve fibers, in which the formation of lamella was clear.

CONCLUSIONBy remitting the impairement of myelinated nerve fibers and enhancing the expression of nNOS in penile tissue, eucommia ulmoides oliv can improve erectile function of diabetic rats.

Animals ; Behavior, Animal ; drug effects ; Diabetes Mellitus, Experimental ; pathology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Eucommiaceae ; Female ; Immunohistochemistry ; Male ; Microscopy, Electron, Transmission ; Nerve Fibers, Myelinated ; ultrastructure ; Nitric Oxide Synthase ; metabolism ; Penis ; drug effects ; enzymology ; innervation ; Random Allocation ; Rats ; Rats, Sprague-Dawley