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Objective: To study the chemical constituents of Ajuga nipponensis., Methods: The chemical constituents were isolated by repeated silica gel column chromatography and their structures were elucidated by phyisochemical properties and spectral analysis., Results: Ten compounds were isolated and identified as:hexadecanoic acid(1), ajuforrestin A(2), beta-sitosterol(3), acacetin(4), apigenin(5), ajugamacrin B(6), ursolic acid(7), beta-ecdysone(8), 8-acetylharpagide(9) and daucosterol(10)., Conclusion: Compounds 1-7 and 10 are isolated from this plant for the first time.

This study was aimed to investigate the correlation between HLA gene distribution and allele frequency of the patients with leukemia. PCR-SSP technique was used to detect the HLA genotype of 2994 umbilical cord blood units from healthy newborns (as control), the detecting result of which was compared with HLA genotypes of 1246 patients with leukemia searched in our cord blood bank. The differences between two groups were compared and analyzed. The results indicated that as compared with the control group, the allele frequencies of HLA-B*56 (0.56%), B*70 (0.24%) obviously increased (RR = 2.2546, 6.2598, χ(2) = 5, 5.98, P < 0.05), while the allele frequencies of HLA-A*03 (3.45%), A*30 (4.86%), B*13 (8.75%), B44* (3.25%), B61* (5.70%), DRB1*07 (8.23%), DRB1*15 (14.21%) obviously decreased in patients with leukemia (RR = 0.5889, 0.7187, 0.7359, 0.5713, 0.7127, 0.6242, 0.7976, χ(2) = 19.23, 9.82, 14.33, 20.48, 11.99, 33.21, 11.56, P < 0.01). It is concluded that HLA-B*56, B*70 alleles seem to be characterized by the genetic susceptibility to leukemia and may be served as risk markers for leukemia occurrence, while the HLA-A*03, A*30, B*13, B*44, B*61, DRB1*07, DRB1*15 can be considered as genetic indicators for resistance of leukemia.

This study was aimed to investigate a quality control method for ABO typing of neonatal umbilical cord blood(UCB). The routine serology method was used to identify the ABO type of UCB samples. These samples with questions were further detected by sequence specific primer PCR (PCR-SSP). The results showed that among total of 76120 UCB samples identified by positive ABO typing, there were 78 samples (1 per thousand) which could not be determined. Of these 78 samples, 30 (56.92%) samples with a weak agglutination reaction were excluded by reverse ABO typing. Out of 260 samples in reverse ABO typing, 148 samples were consistent with positive ABO typing, 112 samples (43.08%) were inconsistent with the positive ABO typing. 58 undetermined samples were detected by PCR-SSP. Out of them the genotyping results of 45 samples confirmed the serological typing, the phenotyping results in 3 cases were inconsistent to that of genotyping. 10 cases showed the unconformity between positive and reverse typing, but the genotyping results were fully consistent with the positive typing. In conclusion, positive typing for red cell antigens combined with PCR-SSP is efficient and sensitive for quality control of ABO typing for neonatal UCB.

This study was purposed to investigate the frequencies of HLA-Cw* loci in China Northern Han population at gene level and to analyze the population genetic characteristics of HLA-Cw* alleles and distribution difference of gene frequency in regions. The high resolution genotyping for HLA-Cw* loci of 420 cases in China Northern Han population was performed by using PCR-SSP typing technique and their distribution regularity was analyzed statistically. The results showed that 30 HLA-Cw* alleles were detected, among which the frequency of Cw* 0102 (0.1776), 0702 (0.1217), 0602 (0.1150) were highest; other alleles with higher frequency were as follow in proper order: Cw* 0304, 0801, 0303, 0302, 0401, 1402. The rare observed HLA-Cw* 0506, 0810, 1510, 1601 and 1701 were detected firstly in this population. The statistical analysis indicated that the genotype distribution of HLA-Cw* loci coincides with the Hardy-Weinberg test. In conclusion, application of high resolution allele typing can accurately understand the distribution regularity and characteristics of HLA-Cw* alleles in China Northern Han population which provides the basis for research related with HLA-Cw* loci.

To unravel the relation of HLA-DRB1*15 with childhood acute lymphoblastic leukemia (ALL), 162 childhood patients with ALL were selected for this investigation. 1 000 normal umbilical cord blood samples were used as control.HLA-DRB1*15 and HLA-DRB5* were typed by polymerase chain reaction (PCR) analysis. The relation of HLA-DRB1*15 with childhood ALL was studied by calculating the chi-square test and relative risk. The results showed that the antigen frequencies and allele frequencies of HLA-DRB1*15 in childhood patients with ALL were 40.12% and 22.62% respectively, while the antigen frequencies and allele frequencies of HLA-DRB1*15 in control were 30.80% and 16.81% respectively, there were significant difference between them (chi(2) = 5.560, p = 0.018, RR = 1.506). In conclusion, the antigen frequencies and allele frequencies of HLA-DRB1*15 in childhood patients with ALL were higher than those in control, so the HLA-DRB1*15 gene is one of the genetic risk factors for childhood ALL. These preliminary data may be useful for further study on the pathogenesis of childhood ALL.

The aim of this study was to investigate the factors which affect HLA typing in 311 umbilical cord blood (UCB) samples. The HLA low resolution typing of UCB samples with misinterpreted HLA types from 311 UCB samples analyzed by PCR-SSO and PCR-SSP was performed. 7 samples difficult to determine their HLA genotype were sequenced directly and the reason leading to misinterpret HLA typing was analyzed. The results indicated that 99.4% of misinterpreted samples resulted from the restriction of HLA typing method itself and 0.6% of misinterpreted samples were suspected to be contaminated with maternal blood in UCB. It is concluded that HLA typing is mainly affected by the shortcomings of oligonucleotide probe design for PCR-SSO and lack of allele specific primers of PCR-SSP.

To investigate the correlation between the HLA genes and pathogenesis of aplastic anemia (AA), polymerase chain reaction with specific sequence primers (PCR-SSP) method was used to HLA typing in 82 patients with AA and 400 normal healthy individuals as control. The results showed that A*2301 (1.84%), B*5501 (4.36%) and DRB1*0901 (23.48%) gene frequency in AA patients were significantly higher than those in controls (relative risk: RR=5.0253, 3.3645, 2.1269, chi2=4.6634, 6.3120, 9.1511 respectively) (p<0.01). In contrast, DRB1*1301 (1.23%) gene frequency was significantly lower in AA than that in controls, RR=0.2257, chi2=6.6629 (p<0.01). It is concluded that A*2301, B*5501 and DRB1*0901 genes may be considered as the risk markers while DRB1*1301 gene as a protective marker of AA.