0.05) compared with the results of PCR ASP genotyping.The consistency between FCM and PCR ASP genotyping was 88% for positive results,100% for negative results,with a total of 95.9%.Conclusion Flow cytometric HLA B 27 typing is rapid and sensitive and can be used together with PCR technique for diagnosis of some diseases such as ankylosing spondylitis.

Objective To understand the molecular genetic basis of Ael subgroup of ABO blood group system in the Han nationality.Method 2 Ael individuals were defined by standard blood group serological techniques,and genomic DNA was prepared for PCR SSP genotyping.Primers were designed and synthesized to amplify complete exon 6 and 7 including flanking intron sequence,and direct sequencing of gel purified PCR amplified fragments was performed using Bigdye Sequencing kit.Result A possibility of regarding the Ael allele as A2,B,O1 and O2 genes had been eliminated by the PCR SSP assay.According to the sequence analysis,Ael gene had 2 mutations of which one was a nucleotide substitution at position 532 in intron 5 (C to T),and the other was a single nucleotide(G) insertion at position 798 to 804 in exon 7 which alter the 86 amino acids sequence of the glycosyltransferase and furthermore extend the translated proteins by 37 amino acids compared with A1 allele.Conclusion The mutations of (789 804)G insertion and C(I 5/532)T substitution is the molecular genetic basis for Ael phenotype.

To observe the effects of Feiyanning (FYN) formula, a compound traditional Chinese herbal medicine, on the expressions of CXCL12 and CXC chemokine receptor 4 (CXCR4) mRNAs and proteins in Lewis tumors in C57 mice.

ObjectiveTo evaluate whether oral administration of Chinese tradiational medicine,Qing-Xue granula,can prevent mouse lung injury caused by thoracic radiation.Methods128 BalB/C mice were divided into 4 groups:control (C) group; radiation (R) group; radiation plus high dose Qing-Xue granula (H) group and radiation plus median dose Qing-Xue granula ( M ) group.The H and M groups were fed 0.64 g and 0.32 g of Qing-Xue granula dissolved in 0.5 nl anline once daily for two months,which were 4 and 2 times of human dosage,respectively.Whole thorax radiation of 12 Gy was delivered with a single ventral-dorsal field with 6 MV X-ray.Group C and group R received 21 days of 0.5 ml saline feeding.Mice were sacrificed at 1,2,4 or 6 months after radiation. Macrophage cell count of lung lavage fluid and hydroxyproline content of left lung were assayed,and the lung fibrosis was scorred according to the Ashcroft's criteria.The plasma interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) concentration were assayed with ELISA method.The One-way ANOVA was used to test the significance of any differences between groups at each time point. Results The macrophage cell number of lung lavage fluid was significantly lower in the 1st month in group M than in group R (2∶4,q =3.92,P ＜ 0.05 ),but had no significant difference between group M and C ( 1 ∶ 4,q =2.13,P＞0.05 ).The hydroxyproline content of group H was significantly lower than group R in the 1st and 6th months (q =3.62,3.54,all P ＜ 0.05 ),but still higher than group C ( q =4.09,3.72,all P ＜ 0.05 ).The fibrosis score of group H was significantly lower than group R in the 2nd,4th and 6th months (q=3.38 -4.16,all P＜0.05).The IL-6 concentration of group H was significantly lower than group R in the 1st month ( q=3.53,P＜0.05 ),but not significantly higher than group C (q =1.41,P＞0.05).The VEGF concentration was significantly higher in group R than group C since the 2nd month ( q =3.12 - 3.78,P ＜ 0.05 ).The VEGF concentration was significantly higher in group H and M than group R in the 2nd and 6th months ( q =3.08 - 3.92,all P ＜ 0.0 5 ).Conclusions Oral Chinese traditional medicine,Qing-Xue granula,could prevent radiation induced lung fibrosis in mice,especially at high dosage.The degree of elevation of VEGF in plasma was not parallel with that of lung fibrosis.

Objective To detect fetal RhCcEe genotype from fetal DNA in maternal plasma for noninvasive prenatal diagnosis.Methods DNA from maternal plasma sample was extracted by use of QIAamp DNA Kit. The existence of fetal DNA was confirmed by amplified fetal SRY gene. The fetal RhCcEe gene was amplified by polymerase chain reaction (PCR) from 30 pregnant maternal plasma. The results of fetal RhCcEe genotype were evaluated retrospectively by the serologic analysis of infant and pregnant woman RhCcEe phenotype.Results Among the 30 samples, 13 were the same phenotypes between mother and infant, 17 were different. When mother phenotypes were RhCC, cc, EE and ee homozygous, the deleted allele gene can be successfully amplified from mother plasma.Conclusion Noninvasive fetal RhCcEe genotyping is reliable. When the mother was homogyzous, genotyping the fetal CcEe alleles was very significant and useful for HDN (hemolysis disease of newborn) diagnosis and therapy.

ObjectiveTo evaluate the effect on lung injury of gefitinib or/and radiation.Methods Totally 160 mice were divided into five groups:control (C) ;gefitinib (G) ;radiation (R) ;gefitinib followed by irradiation ( G + R) ;and R + G.12 Gy irradiation was delivered.Geiitinib fed by 200 mg/kg once daily for 3 weeks.Mice were sacrificed on 1,2,4 or 6 months after radiation.Macrophage count of lung lavage fluid and hydroxyproline assessed,lung fibrosis scored.and plasma TGF-β1 concentration assayed.One-way ANOVA was used to test the significance. Results The lung lavage macrophage cell number were significantly higher in group R,R + G and G + R than group C ( q =2.95 - 8.61,all P ＜ 0.05 ) on 4 and 6months,yet no significant difference between the three groups ( q =0.37 -3.49,all P ＜ 0.05 ) ; The macrophage was significantly lower in month 1,4 and 6 in group G than R,R + G and G + R ( q =3.37- 6.25,all P ＜ 0.05 ).The hydroxyproline content and the fibrosis score of G,R,R + G and G + R were significantly higher than C ( q =3.14 - 4.76,all P ＜ 0.05 ),but no significant difference between the four groups ( q =0.70 - 4.19,all P ＞ 0.05 ).The TGF-β1 concentration of R,G + R,R + G at all time points and TGF-β1 concentration of G at 1 st and 2nd months were significantly higher than C ( q =3.76 -8.09,all P ＜ 0.05).ConclusionsGefitinib could cause lung fibrosis in vivo in BalB/C mouse.The combination of gefitinib and radiation did not significantly exacerbate lung injury caused byeither alone.The mechanism of lung fibrosis caused by gefitinib might be different from that by radiation which needs further research.

Objective:To evaluate the efficacy and safety of oral semaglutide versus sitagliptin in Chinese patients with type 2 diabetes mellitus(T2DM) inadequately controlled with metformin. Methods:The PIONEER 12 study was a phase Ⅲ clinical trial. Chinese patients were prospectively randomized to oral semaglutide(3mg, 7 mg, and 14 mg) or sitagliptin 100 mg. The primary endpoint was the change in HbA 1C from baseline to week 26, and the confirmatory secondary efficacy endpoint was the change in body weight from baseline to week 26. Results:Totally 1 084 Chinese participants(mean age 53 years, male 62.2%, mean duration of diabetes 5.5 years, HbA 1C 8.2%, and body weight 74.3 kg) were enrolled. The changes in HbA 1C at week 26 from baseline were -0.9%, -1.4%, and -1.6% for oral semaglutide 3 mg, 7 mg, and 14 mg, respectively, and -0.7% for sitagliptin. Compared to sitagliptin, oral semaglutide 3 mg, 7 mg, and 14 mg significantly reduced HbA 1C [estimated treatment difference(ETD), -0.2%(95% CI -0.4--0.0), -0.8%(95% CI -0.9--0.6), and -0.9%(95% CI -1.1--0.8), respectively; 3 mg, P=0.011, 7 mg and 14mg, P<0.001]. The estimated mean changes in body weight at week 26 from baseline were -1.1 kg, -2.5 kg, and -3.4 kg for oral semaglutide 3 mg, 7 mg, and 14 mg, respectively, and -0.4 kg for sitagliptin 100 mg. Compared with sitagliptin, oral semaglutide 3 mg, 7 mg, and 14 mg significantly reduced body weight [ETD, -0.8 kg(95% CI -1.3--0.2), -2.1 kg(95% CI -2.6--1.6), and -3.0 kg(95% CI -3.5--2.5), respectively; 3 mg, P=0.004, 7 mg and 14 mg, P<0.001]. The overall incidence of adverse events was similar across all treatment groups. The most common adverse events were gastrointestinal disorders, mostly mild or moderate in severity and transient in duration. Conclusions:Oral semaglutide resulted in significantly greater reduction in HbA 1C and body weight versus sitagliptin at week 26, with a favorable safety and tolerability profile in Chinese T2DM patients inadequately controlled with metformin.

OBJECTIVETo explore the molecular basis of an individual with Bel variant of the ABO blood group.

METHODSThe ABO antigen and serum antibody of the individual were detected by serological method. All coding regions and flanking introns of the ABO gene were amplified with PCR and sequenced bidirectionally. The haplotypes of the individual were analyzed by cloning and sequencing. A three dimensional model of the mutant protein was constructed and analyzed.

RESULTSThe individual has expressed a very weak B antigen on its red blood cells by absorption and elution testing, which was identified as a Bel variant phenotype. The heterozygous sites in exon 6 (261del/G) and exon 7 (297A/G, 484del/G, 526C/G, 657C/T, 703G/A, 796C/A, 803G/C, 930G/A) of the coding region of the ABO gene were identified by direct sequencing. Haplotype analysis showed that the individual has carried an O01 allele and a novel B allele. The sequence of the novel B allele was identical to B101 except for a del G at nucleotide position 484 (484delG), which was nominated as B120 by the Blood Group Antigen Gene Mutation Database (dbRBC NCBI). The 484delG mutation of the B allele has led to a reading frame shift and created a premature terminal codon for the glycosyltransferase (GT) enzyme. Prediction of the 3D structure suggested that the GT enzyme has become an incomplete protein only with its N-terminal region.

CONCLUSIONThe 484delG mutation of the glycosyltransferase B gene has probably abolished or reduced the enzymatic activity and resulted in the Bel variant phenotype.

ABO Blood-Group System ; genetics ; Alleles ; Base Sequence ; Exons ; Female ; Genotype ; Glycosyltransferases ; genetics ; Humans ; Molecular Sequence Data ; Mutation ; Sequence Deletion

OBJECTIVETo explore the function of a novel nonsense mutation c.1476G>A of ITGB3 gene using an in vitro expression system.

METHODSAn eukaryotic expression vector containing ITGB3 c.1476G>A cDNA was generated by site-directed mutagenesis and transformed into E.coli. Plasmid DNA was extracted and sequenced to confirm the target mutations. Wild-type and mutant recombination plasmids were transfected into Chinese hamster ovarian cancer (CHO) cells by nonliposome method, and the stable expression cells were harvested by G418 screening. The ITGB3 gene mRNA transcription and GPIIIa expression level in CHO cells were detected with real-time quantitative PCR, Western blotting and flow cytometry, respectively.

RESULTSThe eukaryotic expression vectors of wild ITGB3 cDNA and c.1476G>A mutant were successfully constructed. CHO cells with stable expression were obtained after transfection and screening. Compared with the wild-type transfected cells, the amount of CD61 antigen expression was 37% and mRNA transcription level was only 6% in the mutant-transfected cells. Full length GPIIIa protein was found only in the stably wild-type-transfected cells, but not in mutant-transfected cells by Western blotting analysis.

CONCLUSIONThe ITGB3 c.1476G>A mutation can decrease the transcription level and further affect GPIIIa synthesis and CD61 antigen expression.

Animals ; Base Sequence ; Blood Platelets ; cytology ; metabolism ; CHO Cells ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Cricetinae ; Cricetulus ; Humans ; Integrin beta3 ; genetics ; metabolism ; Molecular Sequence Data ; Plasmids ; genetics ; metabolism ; Point Mutation

OBJECTIVETo study the serological characteristics and molecular mechanism for a rare Pk phenotype of the P1Pk blood group system.

METHODSThe blood group of the proband was identified by serological techniques. The coding region and flanking intronic sequences of the β-1,3-N-acetylgalactosyltransferase gene (B3GALANT1) associated with the Pk phenotype were analyzed using polymerase chain reaction sequence-based typing.

RESULTSThe proband was identified as having a rare Pk phenotype including anti-P in her serum. The blood group of her daughter and husband showed a P2 phenotype. The nucleotide sequences of the B3GALANT1 gene of her husband and two randomly-chosen individuals were the same as the reference sequence (GenBank AB050855). Nucleotide position 433 C>T homozygous mutation in the B3GALANT1 was found in the proband, which has resulted in a stop codon at amino acid position 145, which may produce a premature protein capable of decreasing or inhibiting the activity of the β -1,3-N-acetylgalactosyltransferase. The nucleotide position 433 C/T heterozygous in the B3GALANT1 was found in her daughter.

CONCLUSIONThe Pk phenotype resulted from 433 C>T mutation in the B3GALANT1 gene has been identified.

ABO Blood-Group System ; genetics ; Adult ; Base Sequence ; Blood Grouping and Crossmatching ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; N-Acetylgalactosaminyltransferases ; genetics ; Pedigree ; Phenotype ; Point Mutation