Agglutinins ; immunology ; Anemia, Hemolytic, Autoimmune ; Humans ; Immunoglobulin G ; Infant, Newborn

A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
Alkyl and Aryl Transferases ; biosynthesis ; genetics ; Azulenes ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Open Reading Frames ; Polyisoprenyl Phosphates ; Recombinant Proteins ; biosynthesis ; Sesquiterpenes ; metabolism ; Sesquiterpenes, Guaiane ; Thymelaeaceae ; enzymology ; genetics

Adult ; Aged ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; enzymology ; Female ; Humans ; Liver Neoplasms ; diagnosis ; enzymology ; Male ; Middle Aged ; RNA, Messenger ; blood ; classification ; alpha-Fetoproteins ; analysis ; gamma-Glutamyltransferase ; blood ; classification

Orobanche caerulescens is an important medicinal resource in Orobanchaceae. The present study aims to establish methods for determination of acteoside, crenatoside, and total phenylethanoid glycosides in O. caerulescens, and determine the content in 15 samples to evaluate the resource utilization of this medicinal plant. The content of acteoside and crenatoside were quantitatively determined by HPLC, while total phenylpropanoid glycosides was estimated by UV-VIS spectrophotometry. According to the results, the content of acteoside was the highest in O. caerulescens, followed by crenatoside. The contents of acteoside, crenatoside, and total phenylethanoid glycosides were between 1.15% - 15.60%, 0.83% - 4.47%, and 6.78% - 27.43%, respectively, which had significant differences. The acquisition time has great influence on the content of main components of O. caerulescens. The content of phenylethanoid glycosides is higher in the samples which were collected at the flowering stage. The two determination methods were proved to be simple, accurate and reliable, and can be used to evaluate the quality and resource utilization of O. caerulescens.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Glycosides ; analysis ; Orobanche ; chemistry

Objective:To evaluate the expression of CXCR4 and the migration rate of bone marrow stromal CD34+cells in differ-ent risk groups with myelodysplastic syndromes (MDS) using correlation analysis. Methods: Forty MDS patients were divided into low-and high-risk groups based on the International Prognosis Scoring System (IPSS). The former was composed of 20 patients with IPSS<1.5, whereas the latter was composed of 20 patients with IPSS≥1.5. Bone marrow (BM) samples of these patients and 10 nor-mal controls were collected. CD34+cells were separated and purified. The expression of CXCR4 was determined by flow cytometry. The migration rate of CD34+cells on the chemotactic effect of SDF-1αand on the effect of bone marrow stromal cells were measured. Results:The expression rate of CXCR4 was higher in the high-risk MDS group than in the low-risk and control groups (P<0.000 1). No significant differences existed between the low-risk and the control groups (P>0.05). The migration rate of CD34+cells on the ef-fects of SDF-1αand marrow stromal cells were significantly increased in the high-risk MDS group compared with those in the low-risk and control groups (P<0.000 1). Migration rate of CD34+cells on the effect of marrow stromal cells was positively correlated with CX-CR4 expression (P=0.000 1). Conclusion:The CXCR4 expression and migration rates of CD34+cells on the effect of marrow stromal cells are significantly higher in the high-risk MDS group than in the low-risk group. Migration rate has a positive correlation with the CXCR4 expression, which further indicates that MDS is a heterogeneous group of hematopoietic stem cell malignancies. The expres-sion and function of SDF-1 and its receptor CXCR4 differ within each group with various risks. SDF-1 and CXCR4 may be involved in MDS pathogenesis.

Hemorrhoids ; Humans ; Male ; Mucous Membrane

Objective To report the clinical effect of primary percutaneous coronary intervention(PCI)in patients with acute myocardial infarction(AMI).Methods A retrospective study was accomplished on the clinical data of 13 AMI patients who underwent PCI from March 2004 to April 2006.Results The infarct-related artery (IRA)was successfully recanalized by primary PCI for 12 AMI patients,without major complications occurred in these cases during hospitalization.Conclusion Primary PCI should be firstly chosen for treatment of AMI in the hospitals which could carry out PCI.

The major antigen region of E2 gene of Hog Cholera Prevalent Strain (Guangxi Yuling Strain) and Chinese Hog Cholera Lapinised Virus (C-strain) derived from hog and rabbit spleen tissue, was amplified by reverse transcription polymerase chain reaction(RT-PCR) and the nested Polymerase Chain Reaction (nPCR). After the amplified fragments were cloned into the expression vector pPROEX-HTb, the recombinant plasmids pPROEX-GXYL and pPROEX-C were obtained. The insert position, the size and the reading frame were right by PCR, restriction digestion and the sequence analysis. SDS-PAGE indicated that both of the reciepient germs transducted and induced by the recombinant plasmids pPROEX-GXYL and pPROEX-C could express the major antigen region of E2 gene. Western-blot indicated that the expressed antigen protein could be recognized by the positive serum of CSFV.
Blotting, Western ; Cloning, Molecular ; Escherichia coli ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology

The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.

Objective Toexplore the correlation of ApoE gene polymorphism and serum uric acid levels in Ningxia Hui Autonomous Region , China.Methods A case-control study.October 2011 to November 2011, five hundred twenty eight ( 296 male, 232 female ) apparently healthy individuals were studied.Questionnaires and physical examinations were performed by standard operation procedure.Fasting blood was collected for biochemistry testing including serum lipid parameters , uric acid concentration and creatinine levels.The multi-ARMS PCR was applied to determine ApoE genotypes ,and the relation of ApoE genotypes with serum lipid parameters and uric acid levels were analyzed.Non-normal distribution were compared using cause and inspection.Results The common six kinds of ApoE genotype can be detected.The total cholesterol ( TC) ,low density lipoprotein cholesterol ( LDL-C) and uric acid ( UA) levels in different genotype subgroups had statistical differences.The individuals with ε2/3 genotype had a significantly greater reductions in TC and LDL-C levels but increment in uric acid concentration than those withε3/3 and ε3/4 genotype (P<0.05).The effect of ApoE gene polymorphism on uric acid levels still remained significantly after adjustment for age , gender , region and other factors.Conclusion The ApoE polymorphism is associated with serum uric acid levels and individuals with ε2 allele have higher serum uric acid levels.