To investigate the molecular genetics basis for one para-Bombay phenotype, the red blood cell phenotype of the proband was characterized by standard serological techniques. Exon 6 and 7 of ABO gene, the entire coding region of FUT1 gene and FUT2 gene were amplified by polymerase chain reaction from genomic DNA of the proband respectively. The PCR products were purified by agarose gels and directly sequenced. The PCR-SSP and genescan were performed to confirm the mutations detected by sequencing. The results showed that the proband ABO genotype was A(102)A(102). Two heterozygous mutations of FUT1 gene, an A to G transition at position 682 and AG deletion at position 547-552 were detected in the proband. A682G could cause transition of Met-->Val at amino acid position 228, AG deletion at position 547-552 caused a reading frame shift and a premature stop codon. The FUT2 genotype was heterozygous for a functional allele Se(357) and a weakly functional allele Se(357), 385 (T/T homozygous at position 357 and A/T heterozygous at 385 position). It is concluded that the compound heterozygous mutation--a novel A682G missense mutation and a 547-552 del AG is the molecular mechanism of this para-Bombay phenotype.
ABO Blood-Group System ; genetics ; China ; DNA Mutational Analysis ; Female ; Fucosyltransferases ; genetics ; Genotype ; Humans ; Male ; Mutation ; Mutation, Missense ; Pedigree ; Phenotype ; Sequence Deletion

OBJECTIVETo evaluate the expression of HDGF and its implication in patients who undergone radical resection for stage I non-small cell lung cancer.

METHODSImmunohistochemical technique was applied to detect the expression of HDGF in 118 lung cancer tissues and 30 normal lung tissues as control. At the same time, the expression of VEGF and Ki-67 labeling rate of the tumors was evaluated.

RESULTSHDGF expression was observed in all cases, and significantly higher than that in normal lung tissues (52.23 +/- 10.35 vs. 156.73 +/- 70.95, P < 0.01). Expresson of HDGF was closely related to histological classification, and the expression in adenocarcinoma was much stronger than that in squamous cell cancers (P = 0.001), but not related to other clinicopathological factors. VEGF expression was closely related to the expression of HDGF. HDGF expression in the VEGF high expression group was much higher than that in VEGF low expression group (171.77 +/- 81.07 vs. 142.81 +/- 59.84, P = 0.028). Ki-67 expression was also closely related to the expression of HDGF, the labeling rate of Ki-67 in high HDGF expression group was much higher than that in low HDGF expression group (30.49% +/- 7.88% vs. 17.80% +/- 5.63%, P = 0.001). Univariate analysis showed that the patients with high HDGF expression had a shorter overall survival than that with low HDGF expression (40.0% vs. 77.5%, P = 0.008), and multivariate Cox regression analysis showed that HDGF was a significantly independent predictive factors for patients with stage I NSCLC (RR = 1.011, P = 0.002).

CONCLUSIONHDGF expression is upgraded in postoperative stage I non-small cell lung cancer patients. HDGF is a significantly independent predictive factor for patients with stage I NSCLC. HDGF may play an important role on carcinogenesis and development of stage I NSCLC through promoting cell proliferation and neoangiogenesis of the tumor.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adult ; Aged ; Biomarkers, Tumor ; analysis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; surgery ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Cell Proliferation ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Ki-67 Antigen ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Staging ; Pneumonectomy ; methods ; Proportional Hazards Models ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism

RHD gene has different alternative transcripts. This study was aimed to construct expression vector of normal mRNA, DEL9 and DEL89 transcripts from RHD gene. Total RNA was extracted from Rh(D) positive umbilical blood cells of newborn. Intact RhD cDNA, DEL9 and DEL89 transcripts were obtained by one-step and two-step RT-PCR, respectively. The obtained products were cloned into pCR4 TOPO sequencing vector for choosing the right transcript. RHD gene was amplified again from the sequencing plasmid DNA, and then subcloned into pcDNA3.1/V5-His TOPO expression vector; DEL9 and DEL89 were cloned into the expression vector directly. Gene sequence and direction were identified by sequencing. The results showed that the sequence and direction of target genes were right, thus these 3 different expression vectors were correctly constructed. It is concluded that expression vector is constructed from different transcripts of RHD gene, which lays a foundation for further exploring the membranous protein expression of Rh(D) antigen.
Cloning, Molecular ; Gene Expression ; Genetic Vectors ; Humans ; Molecular Sequence Data ; RNA, Messenger ; genetics ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

OBJECTIVETo establish a methods based on high-performance liquid chromatogram-mass spectrum for measuring the plasma concentration of nolatrexed dihydrochloride and investigate the pharmacokinetic profile and absolute bioavailability of the drug in mice.

METHODSNolatrexed dihydrochloride were injected intravenously at 50 mg/kg or administered orally at 200 mg/kg in mice, and blood samples were collected at various time points following drug administration. The plasma concentration of nolatrexed dihydrochloride in mice was determined using high-performance liquid chromatogram-mass spectrum. The pharmacokinetic parameters were calculated using DAS software, and the absolute bioavailability of orally and intravenously administered was assessed according to the ratio of their area under the curve (AUC).

RESULTSThe method showed good linear relationship within the drug concentration range of 0.01-40 mg/L (r=0.9995, P<0.001). The recovery of nolatrexed dihydrochloride from the mouse plasma was more than 85%, and the intra- and inter-day precision expressed as the relative standard deviation was less than 15%. The half-life (T(1/2)), AUC, distribution factor and plasma clearance (CL) for intravenously administered nolatrexed dihydrochloride (50 mg/kg) were 3.020-/+0.017 h, 89.972-/+0.425 mg/L/h, 0.831-/+0.106 L/kg, and 0.556-/+0.093 L/h/kg, respectively. The T(1/2), AUC, peak time (T(max)) and peak concentration (C(max)) for orally administered drug were 5.046-/+0.191 h, 84.893-/+9.923 mg/L/h, 1.000-/+0.012 h, and 18.000-/+0.0140 mg/L, respectively. The absolute bioavailability of nolatrexed dihydrochloride in mice was 23.58%.

CONCLUSIONThe absolute bioavailability of nolatrexed dihydrochloride in mice determined in this study provides an experimental basis for development of the oral preparation of the drug.

Animals ; Antimetabolites, Antineoplastic ; blood ; pharmacokinetics ; Biological Availability ; Chromatography, High Pressure Liquid ; methods ; Male ; Mass Spectrometry ; methods ; Mice ; Mice, Inbred C57BL ; Quinazolines ; blood ; pharmacokinetics

OBJECTIVETo evaluate the value of deep small-bowel endoscopy (DSBE) in the diagnosis of Crohns disease (CD).

METHODSThe endoscopic and clinical data of 54 patients with CD receiving capsule endoscopy (CE) and double-balloon enteroscopy (DBE) between January, 2004 and December, 2008 were summarized and analyzed retrospectively.

RESULTSThe main indications for DSBE in our series were suspected CD (42.6%) and obscure gastrointestinal bleeding (25.9%). DSBE was obviously superior to barium imaging. The detection rate of CD was significantly higher with DSBE (92.6%) than with ileocolonoscopy (75.9%, P=0.017), and DSBE provides much more detailed descriptions of specific endoscopic features such as segmental distribution and lumen changes. DSBE significantly improve the diagnostic efficiency, giving priority to offer a guide and raise suspected diagnosis for CD.

CONCLUSIONDSBE is a valuable modality for detecting CD lesions in the jejunum and ileum and for evaluating lesion involvement and severity. The combination with a comprehensive analysis of routine imaging findings, gastro endoscopy, and clinical data can further enhance the diagnostic efficiency of DSBE.

Adolescent ; Adult ; Capsule Endoscopy ; Crohn Disease ; diagnosis ; pathology ; Double-Balloon Enteroscopy ; Female ; Humans ; Intestine, Small ; pathology ; Male ; Retrospective Studies ; Young Adult

OBJECTIVETo establish an in vitro homogeneous time-resolved fluorescence immunoassay method for high throughput screening of protein tyrosine kinase (PTK) inhibitors.

METHODSSpecific fluorescence signals at 670 and 612 nm were measured by multifunctional microplate reader when the fluorescence was emitted through a resonance energy transfer between fluorescent materials (EuK and XL-665). The inhibitory activity of Sunitinib, a standard PTK inhibitor, on vascular endothelia growth factor receptor 2 (VEGFR-2) kinase activity was investigated.

RESULTSA homogeneous time-resolved fluorescence immunoassay was established for high throughput screening of PTK inhibitor. In this system, the concentrations of VEGFR-2, adenosine triphosphate (ATP) and poly-peptide substrate were 5 ng/microl, 100 micromol/L and 1 micromol/L, respectively. Sunitinib inhibited VEGFR-2 kinase activity with an IC50 value of 86.7 nmol/L, which was close to the values tested using other methods.

CONCLUSIONThe homogeneous time-resolved fluorescence immunoassay we established can be easily used for high throughput screening of PTK inhibitors.

Fluoroimmunoassay ; methods ; High-Throughput Screening Assays ; methods ; Indoles ; pharmacology ; Peptides ; metabolism ; Phosphorylation ; drug effects ; Protein Kinase Inhibitors ; pharmacology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; metabolism ; Pyrroles ; pharmacology ; Time Factors ; Vascular Endothelial Growth Factor Receptor-2 ; antagonists & inhibitors ; metabolism

OBJECTIVEAlthough HIV-1 infection is prevalent in many regions in China, it remains largely unknown on the biological characteristics of dominant circulating isolates. This study was designed to isolate the circulating viral strains from different prevalent regions and to characterize their biological properties and neutralization sensitivity.

METHODSPrimary viruses were isolated from fresh PBMCs using the traditional co-culture method and their capacity of inducing syncytium was tested in MT-2 cells. Meanwhile, their coreceptor usage was determined with two cell lines: Magi and GHOST (3) stably expressing CD4 and the chemokine receptor CCR5 or CXCR4. Furthermore, the sensitivity of these viruses to neutralization by HIV-1-infected patients' plasma which were highly active to neutralize SF33 strain, was quantified in GHOST cell-based neutralization assay.

RESULTSSix primary viral strains were isolated from 4 separated regions. Isolates LTG0213, LTG0214 and XVS032691 induced syncytia in MT-2 cells, and used CXCR4 as coreceptor. Isolates XJN0021, XJN0091, or SHXDC0041 did not induce syncytia, and used CCR5 as coreceptor. Overall neutralization sensitivity differed among four representative strains: HIV-1 XVS032691 > LTG0214 >XJN0091 approximately SHXDC0041.

CONCLUSIONThe neutralization sensitivity of HIV isolates is linked with the phenotype of isolates, in which syncytium-inducing (SI) or CXCR4-tropic (X4) viruses are more easily neutralized than non-syncytium-inducing (NSI) or CCR5-tropic (R5) viruses. The genetic subtypes based on the phylogeny of env sequences are not classical neutralization serotypes.

CD4-Positive T-Lymphocytes ; metabolism ; Cell Line ; Cells, Cultured ; Chemokines ; genetics ; immunology ; China ; Coculture Techniques ; methods ; Giant Cells ; ultrastructure ; virology ; HIV Infections ; virology ; HIV Seropositivity ; genetics ; immunology ; HIV-1 ; immunology ; isolation & purification ; physiology ; Humans ; Neutralization Tests ; Receptors, CCR5 ; metabolism ; Receptors, CXCR4 ; metabolism ; Virus Replication

OBJECTIVETo study the present situations of lung cancer in Wuhan and to explore the relationship between the potential years of life lost of lung cancer and air pollution, especially vehicle emissions.

METHODSData gathered between 1986 and 1995 in Wuhan city, including air pollution and tobacco production and data on lung cancer between 1991 and 2000 were collected extensively. Simple Correlation and Grey Relational Analysis were used to analyze the relationship of them.

RESULTSThere was a ascending tendency in variance of oxides of nitrogen (NOx). The degree of grey incidence (DGI) between the concentration of air pollutants and the male's or female's potential years of life lost of lung cancer (PYLL) were calculated respectively. In males, the values of DGI were 0.6702, 0.7071, 0.6199 on sulfur dioxide (SO2), NOx, total suspensions (TSP) respectively. In females,the values of DGI were 0.6188, 0.8555, 0.5842 according to the same order as listed above. Significant positive correlation was found between the concentration of NOx and with lung cancer in both males and females by spearman correlation test (rmale = 0.63523, P = 0.0484; rfemale = 0.76396, P = 0.0101).

CONCLUSIONWith the fast growing speed of the quantity of vehicles, pollution of vehicle emission-caused air pollution posed an important risk factor for lung cancer, despite the fact that tobacco smoking still played the leading role.

Air Pollution ; adverse effects ; China ; Female ; Humans ; Lung Neoplasms ; mortality ; Male ; Nitrogen Oxides ; analysis ; Sulfur Dioxide ; analysis ; Vehicle Emissions ; analysis

OBJECTIVETo investigate the inhibitory effect of naringin on monocyte adhesion to high glucose-induced human umbilical vein endothelial cells (HUVECs).

METHODSCultured HUVECs isolated from human umbilical cords were pretreated with or without naringin and induced with high glucose (33 mmol/L) for 48 h. Human monocyte THP-1 cells, after labeling with BCECF-AM, were co-cultured with the HUVECs for 30 min. The labeled THP-1 cells adhering to HUVECs were observed under fluoroscence microscope, and the inhibitory effect of naringin on the cell adhesion was evaluated by measuring the adhering cell density. Western blot analysis was used to detect the expressions of the adhesion molecules in the HUVECs, and reactive oxygen species (ROS) production in the HUVECs was measured using an oxidation-sensitive fluorescent probe (DCFH-DA). The nuclear extracts of the HUVECs were prepared to examine the expression of nuclear factor-kappa B (NF-kappaB) in the cell nuclei by Western blotting.

RESULTSHUVECs in high-glucose culture showed increased adhesion to THP-1 cells and enhanced expressions of the cell adhesion molecules, which were significantly attenuated by pretreatment with naringin (10-50 microg/ml). High glucose induced DCF-sensitive intracellular ROS production in the HUVECs, and this effect was inhibited by naringin pretreatment of the cells. Naringin also suppressed high glucose-induced increment of NF-kappaB expression in the cell nuclei of HUVECs.

CONCLUSIONNaringin can suppress high glucose-induced vascular inflammation possibly by inhibiting ROS production and NF-kappaB activation in HUVECs.

Cell Adhesion ; drug effects ; Cell Adhesion Molecules ; metabolism ; Cell Line ; Cells, Cultured ; Coculture Techniques ; Endothelial Cells ; cytology ; Flavanones ; pharmacology ; Glucose ; antagonists & inhibitors ; pharmacology ; Humans ; Monocytes ; cytology ; NF-kappa B ; metabolism ; Reactive Oxygen Species ; metabolism ; Umbilical Veins ; cytology

OBJECTIVETo compare the in vitro inhibitory effect of expolysaccharides from Streptomyces, polysaccharides of Ganoderma lucidum and rice bran on six-alpha-helix bundle formation of HIV gp41 protein.

METHODSThe amount of six-alpha-helix bundle formed in the presence of N36 and C34 was tested by ELISA in response to treatments with different doses of polysaccharides.

RESULTSExpolysaccharides from Streptomyces potentially inhibited six-alpha-helix bundle formation with the effective concentration (IC(50)) of 145.48-/+7.25 mg /L. Polysaccharides of Ganoderma lucidum and rice bran showed no effect on the six-alpha-helix bundle formation.

CONCLUSIONExpolysaccharides from Streptomyces can inhibit the six-alpha-helix bundle formation of HIV gp41, whereas polysaccharides of Ganoderma lucidum and rice bran do not exhibit such activity.

HIV Envelope Protein gp41 ; chemistry ; Kinetics ; Oryza ; chemistry ; Polysaccharides ; pharmacology ; Protein Structure, Secondary ; drug effects ; Reishi ; chemistry ; Streptomyces ; chemistry