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 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

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 study the impact of TNM staging and combined treatment mode on the survival of non-small cell lung cancer (NSCLC) patients.

METHODSFrom January 1997 to December 2002, 987 NSCLC patients were surgically treated in this hospital. Of those, 574 received combined modality therapy (surgery + chemotherapy/radiotherapy), while 413 underwent operation alone. Their clinicopathological data were retrospectively analyzed.

RESULTSThe 1-, 3-, 5-, and 10-year overall survival rates were 87.7%, 57.5%, 54.6% and 54.5%, respectively, for the whole group, which were 90.6%, 57.5%, 54.3% and 54.1% for the combined therapy group versus 83.8%, 57.6%, 55.2% and 55.2% for the group treated by surgical resection alone. The 1-year survival rate of the combined therapy group was significantly higher than that of the surgical resection alone group (90.6% vs. 83.8%) (P<0.01). With regard to the T factor, 5-year survival rate of the combined therapy group (surgery + radiotherapy) was higher than that of surgery alone group, especially in T4 cases (43.6% vs. 12.7%), with a significant difference between them (P<0. 05). As for the N factor, the 1-year survival rate of NO patients in the combined therapy group (surgery + chemotherapy/radiotherapy) was significantly higher than that of surgery alone group (94.4%, 97.9% vs. 90.0%) (P<0.05). The 1-year survival rate of N1 patients in the combined therapy group (surgery + chemotherapy or + chemotherapy and radiotherapy) was 91.7% and 100% versus 82.9% in the surgery alone group (P<0.01); The 1- and 3-year survival rates of N2 patients in the combined modality therapy group (surgery + chemotherapy) were 82.1% and 37.3%, while those of the surgery alone group were 69.4% and 26.5%, respectively, with a significant difference (P<0.05, P<0.01). All the severity of primary tumor, distance of lymph node involvement, and distant tumor metastasis significantly worsen the prognosis of the patients.

CONCLUSIONThe prognosis in NSCLC patients treated with combined modality therapy (surgery + chemotherapy/radiotherapy) is better than that with surgery alone. The larger the original tumor and the farther the lymph node and tumor metastases, the worse the prognosis is for NSCLC patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; pathology ; therapy ; Chemotherapy, Adjuvant ; Child ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; pathology ; therapy ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Pneumonectomy ; methods ; Radiotherapy, Adjuvant ; Retrospective Studies ; Survival Rate ; Young Adult

OBJECTIVETo study the surgical management of fatal hemorrhage following head and neck surgery for cancer.

METHODSThe clinical data of 32 cases of fatal hemorrhage following head and neck surgery from 1976 to 2008 in our department were analyzed retrospectively.

RESULTSHemorrhage was caused by carotid blowout in 20 cases. The carotid ligation was performed in 13 cases, only 6 cases got long-term survival. In 12 cases, hemorrhage was caused by tracheo-innominate artery fistula, only 2 cases received surgical management, and no long-term survivors.

CONCLUSIONFatal hemorrhage following head and neck surgery is an uncommon but frequently fatal complication, and the successful management of it depends on early diagnosis and correct treatment.

Adult ; Aged ; Carotid Artery, Common ; surgery ; Female ; Head and Neck Neoplasms ; pathology ; radiotherapy ; surgery ; Humans ; Laryngectomy ; adverse effects ; methods ; Ligation ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck Dissection ; Postoperative Hemorrhage ; etiology ; surgery ; Retrospective Studies ; Young Adult

Factor Analysis, Statistical ; Humans ; Tuberculosis ; mortality

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