0.05),and during which,no adverse drug reactions were observed.The prophylactic medication days,antibiotic drugs costs and total drugs costs in hospitalization were significantly lower in the trial group than in the control group(P

The authors determined the concentration of matrine in rabbit blood plasma with HPLC after it was injected intravenously at constant rate and studied the pharmacokinetics with following five dosage groups of 10,15,20,30 and 60 mg/kg,respectively.As a result,the pharmacokinetics of matrine in rabbit body coincides with two-compartment open model.For 10-30mg/kg dosage groups,the t_(1/2)?,Vc and CLS are similar basically. A direct proportion increase in AUC as dose is increased.The matrine reveals linear elim- ination.For 60 mg/kg dosage group,t_(1/2)? obviously lengthens.AUC increases over pro- portion.The elimination of matrine is nonlinear.

In this paper, the biological compatibilities of FGF/Collagen compound sponges are evaluated. Intracutaneous stimulation test and shortterm muscle embedding test of FGF/Collagen compound sponges made of different materials are performed. Experimental data are analyzed and evaluated according to the standard. The two sponges get marks of 0 in acute eye stimulation test and intracutaneous stimulation test, thus they are not stimulating to tissues. In shortterm muscle embedding test, they don't lead to inflammation reactions and can be degraded and absorbed in short term. FGF/Collagen compound sponge proves good biological compatibility and has a cheerful prospect in medicine.

le full matching for unrelated donor-receipt pairs. However, HLA-Cw mismatching at antigen level could no longer be ignored. The results indieated that HLA-Cw genotyping should be incorporated into future CMDP unrelated marrow donor routine HLA test.

Objective To establish a method for the determination of protocatechuic acid, salidroside, and chlorogenic acid in Sargentodoxa cuneata. Methods The separation was performed on a Waters XSELECT CSH C18 (150 mm × 4.6 mm, 5 μm) with methanol-acetonitrile-0.2 % phosphoric acid as the mobile phase in a gradient elution at a flow rate of 0.8 ml/min. The detection wavelength was 260 nm and the column temperature was 35 ℃. Results The linear ranges of protocatechuic acid, salidroside, and chlorogenic acid were 0.0020-0.0120, 0.0600-0.3602, 0.0750-4.5006 mg/ml, respectively. The average recoveries were 98.01% (RSD=0.07%), 98.53 % (RSD=0.12%), and 101.10 % (RSD=1.92%), respectively. Conclusions The method is simple, accurate, and highly reproducible, which could provide the scientific evidence for the quality control of Sargentodoxa cuneata.

Objective To study the molecular genetic polymorphism of exons 1,5, 6, 7 of HLA-C gene in Chinese population and evaluate the significance of additional sequencing based typing at exons 1,5, 6, 7 of HLA-Cw gene in clinical HLA matching, Methods A total of 324 individuals were typed at exons 2,3, 4 of HLA-C gene by sequence-based typing. If ambiguities appeared outside of exons 2 -4, we designed a total of 5 in-house sequencing primers and optimized the sequencing reaction, additional sequencing based typing at exons 1,5, 6, 7 was performed to solove the emerging ambiguities. Results In the three hundred and twenty-four samples typed by PCR-SBT at exons 2, 3 and 4 of HLA-Cw gene, 23.8 % (77/324) of the typed samples were assigned the conclusive genotype in four digital level 76. 2% (247/324) of the typed samples were given with the ambiguous allele combination results, in which 73 kinds of ambiguous allele combinations were detected. Increasing the additional sequencing analysis at exons 1, 5, 6, 7 of HIA-C gene, ten frequent ambiguities including Cw* 030201/030202, Cw* 070201/0750, Cw* 040101/0409N/0430, Cw* 0403/0409N/0430, Cw* 080101/0822 could be distinguished. ConclusionsIncreasing the sequencing anlysis at exons 1, 5, 6 and 7 of HLA-Cw gene will help to make clear the ambiguous SBT results and also improve the accuracy of HLA-Cw typing. It shows important significance in clinical histoeompatibility matching.

Objective To study the molecular genetic background of Bel subtype at ABO blood group.Methods Three samples and fifteen samples were diagnosed as Bel subgroup and normal control samples by serological test,respectively.The extracted DNA was genotyped by sequence specific primer- polymerase chain reaction foilowed by sequencing for Exon6 and exon7 at ABO locus and clones were sequenced.Results A novel Bel variant allele(GenBank EF117687) was identified in a Bel individual.The Bel allele was different from the regular B101 allele by single 952G>A missense mutation in exon7.resulting in an amino acid subsfitution of Val for Met at 318 locus.No mutations were detected in the fifteen control samples and the other two Bel allele samples.Conclusions The mutation position was fimt found to lie on coding region of ABO gene behind nucleotide 930.The mutation of G952A in the al,3 galactosyhransferase gene may be one of the molecular genetic basis of Bel ohenotype.

OBJECTIVETo explore the association of KIR-HLA gene polymorphism with chronic myeloid leukemia (CML) among ethnic Hans from southern China.

METHODSA total of 172 adult CML patients and 480 unrelated healthy controls were screened for the presence of KIR with sequence-specific primers-PCR (PCR-SSP) and sequence-based typing (SBT) of HLA-A, -B and -C loci. Polymorphisms of the KIR-HLA system were analyzed at 4 levels, and the frequencies of KIR framework genes and KIR profiles, classⅠHLA ligands, matched KIR+HLA pairs and KIR-HLA compound profile were compared between the two groups. P values were calculated using SPSS 13.0 software.

RESULTSFor the CML group, the frequencies of HLA-C2 ligand, 2DL1+HLA-C2 pair and HLA-B Bw4-80I were significantly lower than those of the control group, suggesting a protective effect against CML (HLA-C2: OR=0.386, 95%CI:0.240-0.620, P<0.01; 2DL1+HLA-C2: OR=0.316, 95%CI:0.191-0.525, P<0.01; HLA-B Bw4-80I: OR=0.576, 95%CI:0.384-0.862, P<0.01). The frequencies of KIR2DL1 ligand (HLA-C2) and KIR3DL1 ligand (HLA-B Bw4-80I) in the CML group were significantly lower than that of the control group, suggesting that the HLA-C2 and HLA-B Bw4-80I expression is probably decreased in the CML patient group, which led to reduced inhibitory signal and enhanced activating signal of KIR2DL1and/or KIR3DL1NK cells. Notably, the frequency of KIR-HLA compound profiles ID2 (KIR AA1-HLA-C1/C1-Bw6/Bw6-A3/11) in CML patients significantly increased in the CML patient group compared with the control group, suggesting that the KIR-HLA compound profiles ID2 may be a risk factor for CML (OR=2.163, 95%CI 1.198-3.906, P<0.01).

CONCLUSIONAbove analysis has identified certain protective and risk factors for CML from the KIR-HLA system, which may provide a clue for the pathogenesis of leukemia and development of individualized immune therapy.

Asian Continental Ancestry Group ; genetics ; China ; Gene Frequency ; Genetic Predisposition to Disease ; ethnology ; genetics ; Genotyping Techniques ; HLA Antigens ; genetics ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-C Antigens ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; ethnology ; genetics ; Odds Ratio ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Protein Isoforms ; genetics ; Receptors, KIR ; genetics ; Risk Factors

OBJECTIVETo investigate the number and ratio of ambiguous allele combinations from human leukocyte antigen (HLA) confirmatory test by sequencing based typing for unrelated donor marrow transplantation, and to establish an efficient strategy for identifying such ambiguities.

METHODSA total of 650 donor-receipt samples were genotyped for 5 loci of the HLA gene using an Atria SBT commercial kit. Exons 2, 3 and 4 of HLA-A, -B and -C, exon 2 of HLA-DRB1 and exons 2 and 3 of HLA-DQB1 were tested by routine HLA genotyping. The ratio of usual ambiguous allele combination was calculated. The ambiguities were subjected to further confirmatory test by PCR-SSP or PCR-SBT retest at outside of the routine sequencing region.

RESULTSAmong the 650 tested samples, the ratio of ambiguity at HLA-A, B, C, DRB1 and DQB1 were 76.31% (496/650), 91.08% (592/650), 97.69% (635/650), 88.62% (576/650) and 43.38% (141/650), respectively. A total of 36 ambiguous allele combinations inside the routine sequencing region and 22 ambiguous allele combinations outside of the routine sequencing region were discovered. After removing rare alleles based on the Chinese common and well documented (CWD) Allele Table (Version 1.01), 9 ambiguous CWD allele combinations inside the routine sequencing region, including 3 located in HLA-B, HLA-C and 1 located in other three HLA loci were found. Ten ambiguous CWD allele combinations outside of the routine sequencing region, including 4 located in HLA-C, -DRB1 and 1 in HLA-A, -B respectively were determined. All samples with ambiguous CWD allele combinations could be distinguished by high-resolution PCR-SSP commercial kits or PCR SBT retest at outside of the routine sequencing region.

CONCLUSIONThe common and well documented allele combinations in sequencing-based typing at five HLA loci have been analyzed. Our strategy may provide valuable information for more efficient, low cost and accurate method for high resolution genotyping of HLA genes.

Alleles ; Genotype ; HLA Antigens ; genetics ; Humans

OBJECTIVETo explore the reason for HLA-DQB1 allele dropout during routine sequence-based typing(SBT) in order to improve the accuracy of typing.

METHODSTwo thousand samples derived from HLA high-resolution typing laboratory were typed for HLA-DQB1 locus using an AlleleSEQR HLA-DQB1 SBT kit. Non-conclusive results and "abnormal" sequencing samples were retyped using a LABType rSSO HD HLA-DQB1 kit and further analyzed with both sequence-specific primers and group-specific primers and sequenced for haplotype analysis.

RESULTSAmong the 2000 samples, 2 samples with no conclusive result were identified. The heterozygosity was confirmed with both the LAB Type SSO HD HLA-DQB1 kit and PCR-SBT in house method. Subsequent HLA-DQB1 cloning and haplotype sequencing have elucidated that HLA-DQB1*02:02 dropped out at exon 2 for the first sample and HLA-DQB1*02:01:01 dropped out at exon 2 for the second sample during PCR amplification. No novel nucleotide mutation was found.

CONCLUSIONOur results indicated that preferential amplification at exon 2 of DQB1 may result in allele dropout in exon 2 sequences during HLA-DQB1 SBT test. This may provide useful information for HLA genotyping.

Alleles ; DNA Primers ; genetics ; Exons ; Genotype ; HLA-DQ beta-Chains ; genetics ; Histocompatibility Testing ; methods ; Humans ; Polymerase Chain Reaction ; methods