Object To study the chemical constituents from the fruits of Juniperus formosana Hayata. Methods The compounds were isolated and purified on liquid-liquid, silica gel, and aluminium oxide column chromatography, their structures were identified by spectroscopic methods, such as GC-MS, 1H-NMR, and 13C-NMR, especially 2D-NMR. Results Two new oxygenated diterpenoids, named labdatriene (Ⅰ) and labdadiene (Ⅱ), were isolated from the ethyl acetate extract. Their structures were determined to be 19-carboxy-8(17)-13(16)-14-labdatriene and 15, 19-dihydroxyl-8(17)-13(E)-labdadiene, respectively. Conclusion Compounds Ⅰ and Ⅱ are new diterpenoids.

OBJECTIVETo identify two novel HLA alleles HLA-B*9536 and B*4612, in an individual.

METHODSDNA was extracted from whole blood by Invitrogen DNA extraction kit. The amplification for HLA-B exons 2-4 of the proband was performed separately with allele group specific primers and the PCR products were directly sequenced for exons 2-4 in both direction.

RESULTSThere were two novel HLA-B alleles in the proband. The sequences of the two alleles have been submitted to GenBank (EU081878 and EU081879). The two alleles have been officially named as B*9536 and B*4612 by the WHO Nomenclature Committee. The sequence of exons 2-4 of HLA-B*9536 showed one nucleotide difference in exon 3 at position 544 (G to A) comparing with the closest allele B*1505, which resulted in an amino acid change from Ala to Thr at codon 158. In the HLA-B*4612 allele, there was one nucleotide change in exon 3 at position 363 (G to A), when compared to the closest allele B*4601, which lead to an amino acid change from Arg to Ser at codon 97.

CONCLUSIONTwo novel HLA-B alleles were identified in one individual and have been officially named by the WHO Nomenclature Committee.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; HLA-B Antigens ; genetics ; HLA-B15 Antigen ; Humans ; Male ; Molecular Sequence Data

OBJECTIVE: To detect loss of heterozygosity (LOH) at human leukocyte antigen (HLA) loci in a Chinese patient with leukemia after haploidentical hematopoietic stem cell transplantation. METHODS: HLA genotyping was carried out on peripheral blood, hair follicle and buccal swab samples derived from the patient after the transplantation as well as peripheral blood samples from his parents by using PCR-sequence specific oligonucleotide probe method and PCR-sequence based typing method. Short tandem repeat (STR) loci were detected by using a 23 site STR assay kit and a self-developed 6 STR loci assay for the HLA regions. RESULTS: After the transplantation, the HLA genotype of the peripheral blood sample of the patient was identical to his father. The patient was HLA-A*02:01,24:02, C*03:03,03:04, B*13:01,15:01, DRB1*08:03,12:02, DQB1*03:01,06:01 for his hair follicle specimen. However, homozygosity of the HLA loci was found in his buccal swab sample. Only the HLA-A*24:02-C*03:03-B*15:01-DRB1*08:03-DQB1*06:01 haplotype from his father's was present, while the HLA-A*02:01-C*03:04-B*13:01-DRB1*12:02-DQB1*03:01 haplotype from his mother was lost. After the transplantation, the alleles of the 23 STR sites in the patient's peripheral blood sample were consistent to his father, with no allelic loss detected in his buccal swab sample. However, at least 4 STR loci in the HLA region were lost in his buccal swab sample. CONCLUSION LOH at the HLA loci has been detected in the buccal swab sample of a patient with leukemia who received haploidentical hematopoietic stem cell transplantation.
HLA Antigens/genetics* ; HLA-A Antigens/genetics* ; Histocompatibility Antigens Class I/genetics* ; Humans ; Leukemia/genetics* ; Loss of Heterozygosity

Objective:To explore the significance of enthesitis in psoriasis(PsO) and the value of high-frequency Doppler ultrasound (HDUS) in screening potential subclinical psoriatic arthritis(PsA) patients through enthesitis; Designing the semi-quantitative score of enthesitis explore the value of semi-quantitative score in evaluating the degree of joint structure damage and improving the curative effect in patients with subclinical PsA.Methods:Clinical and sonographic data were collected in 125 PsO patients and 50 osteoarthritis (OA) patients who attended hospital from January 2019 to March 2021, and healthy controls (HC) recruited during the same period. The prevalence of enthesitis of distal interphalangeal joint (DIP) of psoriasis patients, osteoarthritis patients and HC was compared. Meanwhile the semi-quantitative scoring system for ultrasound-guided enthesitis was designed, and the correlation between the severity of enthesitis and clinical score was analyzed. The correlation between psoriasis nail and enthesitis of extensor tendon of DIP finger was also analyzed.Results:The significant difference was found in the prevalence of enthesitis of DIP among PsO group, OA group and HC group (all P<0.001); There was a significant difference in the severity of enthesitis of DIP in PsO group before and after treatment ( P<0.001); In PsO group, the correlation between the severity of enthesitis of DIP and Psoriasis Area and Severity Index(PASI)score was moderate ( rs=0.538, P<0.001), and highly correlated with Nail Psoriasis Severity Index (NAPSI) scores( rs=0.877, P<0.001). There was a significant difference in the incidence of enthesitis between the patients with psoriasis nail and the patients without that ( P<0.001). The prevalance of enthesitis of nail disease free fingers was also found significantly different between PsO group and HC group( P<0.001). Conclusions:Enthesitis is a prominent characteristic of PsO.And the semi-quantitative scores designed in this research are in satisfied consistence with the clinical scores, and could be used to evaluate subclinical PsA structural damage.Furthermore, it is possible that the enthesitis of DIP is a relatively independent process from the nail disease.

【Objective】 To statistically analyze the characteristics of ambiguous results of HLA-DPB1 genotyping given by AllType NGS 11-loci sequencing reagent via two next generation sequencing platforms, i. e. Ion Torrent S5 and Illumina Miseq. 【Methods】 A total of 434 samples from patients or donors were genotyped for HLA-DPB1 locus using AllType NGS 11-loci sequencing reagent from One Lambda company; 336 samples of them were sequenced via the Ion Torrent S5 platform and other 98 samples were sequenced via the Illumina Miseq platform. All 434 samples were genotyped for HLA-DPB1 gene simultaneously using PCR-SSO flow fluorescent bead method. The ambiguous genotypes of HLA-DPB1^*13∶01∶01/107∶01 were distinguished by Sanger sequencing. The HLA-DPB1 genotype results by NGS method were assigned by TypeStream Visual professional software, and the ratio of ambiguous combination was calculated by direct count method. 【Results】 Ambiguous results were found in 357 out of 434 samples, accounting for 82.3% (357/434) when HLA-DPB1 allele was assigned to the third field using NGS method. Ambiguous results with 45 types were given in 275 out of 336 samples by the Ion Torrent S5 platform, accounting for 81.8% (275/336) and 82(with 27 types) out of 98 samples by the Illumina Miseq platform, accounting for 83.7% (82/98). All samples were re-genotyped for HLA-DPB1 gene by PCR-SSO, and none HLA-DPB1 allele had been missed by NGS. A total of 43 ambiguous alleles in HLA-DPB1^*13∶01∶01/107∶01 involving 41 samples were distinguished by Sanger sequencing; HLA-DPB1^*13∶01∶01 were detected in 25 (58.1%, 25/43) and HLA-DPB1^*107∶01 in 18 (41.9%, 18/43). 【Conclusion】 There were still a high proportion of HLA-DPB1 ambiguous combinations using the AllType NGS 11-loci sequencing reagent. Sequencing exon 1 of HLA-DPB1 gene by Sanger sequencing can resolve part of the ambiguous results in HLA-DPB1^*13∶01∶01/107∶01 alleles.

OBJECTIVETo detect the expression of Mycobacterium tuberculosis secreted protein Ag85B in paraffin-embedded tissues by immunohistochemistry (IHC), and to evaluate its application in the pathological diagnosis of tuberculosis.

METHODSOne hundred and five tuberculosis specimens (54 pulmonary tuberculosis, 51 lymph nodal tuberculosis) and 51 specimens of other diseases (8 lung cancer, 10 pulmonary abscess, 10 bronchiectasis, 7 lymphoma, 5 necrotizing lymphadenitis, 4 reactive hyperplasia lymphoid, and 7 sarcoidosis) were collected from January 2012 to July 2013 from Beijing Chest Hospital, Capital Medical University. One-step IHC was performed on paraffin-embedded tissues using antibody directed against Ag85B.

RESULTSIHC and Ziehl-Neelsen (ZN) acid-fast staining showed that distribution and intensity of Ag85B expression were concordant with the distribution and number of acid-fast bacilli. IHC showed significantly higher sensitivity than ZN staining (50.5%, 53/105 vs. 31.4%, 33/105; χ² = 7.877, P = 0.005). The combined sensitivity of IHC and ZN staining was 59.0%. Moreover, oil immersion was not necessary for IHC, allowing more rapid diagnosis.

CONCLUSIONIHC detection of Ag85B is a simple method with higher sensitivity than ZN staining, and demonstrated good value in the pathological diagnosis of tuberculosis.

Acyltransferases ; metabolism ; Antigens, Bacterial ; metabolism ; Biomarkers ; metabolism ; Bronchiectasis ; diagnosis ; immunology ; Humans ; Immunohistochemistry ; Lymphadenitis ; diagnosis ; immunology ; Mycobacterium tuberculosis ; immunology ; Sarcoidosis ; diagnosis ; Staining and Labeling ; Tuberculosis, Lymph Node ; diagnosis ; immunology ; Tuberculosis, Pulmonary ; diagnosis ; immunology