Objective To do analysis of sequence and identify a novel HLA-A allele in Chinese hematopoietic stem cell do-nors.Methods A rare HLA-A allele was initially detected by Luminex PCR-SSO typing,then the sample was sequenced by sequence-based typing (SBT)and the group-specific sequencing primer (GSSP)to confirm the mutation allele and locus.Re-sults The sequence of the sample results showed that the allele compared with the highest homologous allele HLA-A*24∶02∶01∶01 was the difference in the exon 3 at position 544 G>A,resulting in an amino acid sequence of HLA-A*24∶02∶01∶01 at position 158 change Ala to Thr.Conclusion This allele is a new HLA-A allele and has been designated as HLA-A*24∶327 by the HLA Nomenclature Committee of World Health Organization (WHO).

Objective To establish an effective and stable rat model of skin-infection for evaluating the therapeutic effects of topical anti-infection drugs.Methods SD rats were subcutaneously injected with cyclophosphamide to induce immunosuppression,and then surgical incisions were made on both sides of the spine.The rat model of skin-infection was established by applying methicillin-resistant Staphylococcus aureas suspension to the incisions for two consecutive days,and evaluated by analyzing infection status,pathological changes and healing time.Results The development of pyogenic infection was detected in all of the rats.Compared with the non-cyclophosphamide treated group,the cyclophosphamide treated group showed a more severe infection both from the visual inspection and the microscopic observation,moreover,its healing time was delayed more than 10 days.Conclusion The skin-infection model was successfully established in immunosuppressed rats induced by cyclophosphamide,which could be applicable to the efficacy evaluation of anti-infection drugs for external use on skin infection.

OBJECTIVETo investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine.

METHODSC57BL/6 mice were randomly divided into 3 groups: experimental group I (treated with pcDNA L1), control group II (treated with pcDNA3.1) and control group III (treated with PBS buffer). The mice were immunized three times during a three-week interval. Ten to fourteen days after the third inoculation, a footpad swelling test was used to detect delayed-type hypersensitivity (DTH) responses. Antigen-specific splenocyte proliferation assay and quantitation of IFN-gamma cells in splenocytes were performed by FACS assay.

RESULTSIn the experimental group, splenocytes actively proliferated after stimulation with HPV16 VLP, and had developed a markedly larger amount of CD8(+) IFN-gamma(+) cells, which is an index for special CTL. Also, the footpad was significantly thickened upon inoculation with HPV16 VLP.

CONCLUSIONNaked DNA vaccine of HPV16 L1 can induce specific cell-mediated immune responses in mice, which should be considered for evaluation of HPV16 DNA vaccine feasibility.

Animals ; Hypersensitivity, Delayed ; etiology ; Immunization ; Interferon-gamma ; biosynthesis ; Lymphocyte Activation ; Mice ; Mice, Inbred C57BL ; Papillomaviridae ; immunology ; Spleen ; cytology ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

Objective It has been well accepted that human papillomavirus type 16 (HPV16) is associated with human cervical cancer and HPV16 L1 protein could induce both humoral and cellular immune responses. The objective of this study is to map epitopes on HPV16 L1 protein and to provide information to the design of HPV16 prophylactic peptide vaccine.Methods The epitopes on L1 protein were screened by polyclonal and 2 monoclonal antibodies (B8 and F4G3) against HPV16 L1 protein from a 6-mer phage display epitope library with the method of immuno-affinity screening (Bio-panning). After 3 rounds of Bio-panning, the positive phages were detected by L1 antibodies again with ELISA. The positive phages reacted strongly with L1 antibodies were then identified by DNA sequencing.Results Three mimotopes have been screened by polyclonal and two monoclonal antibodies. The mimotope (LSLFSC) which reacted with monoclonal antibody B8 showed 50% homology with the sequence 270～275a.a (DSLFFY) of prototype HPV16 L1. Another mimotope (LTSSYS) which reacted with polyclonal antibodies had 66% homology with the L1 sequence 516～521a.a (TTSSTS), also a mimotope (DRWDRF) was found to have the homologic RF with the known L1 sequense 441～446a.a. The mimotopes LSLFSC and DRWDRF were adjacent to the epitopes at 267～269a.a and 422～441a.a reported by other researchers previously.Conclusions Our results suggested that there might be a batch of epitopes on HPV16 L1 protein, and the predominant epitopes of HPV16 L1 protein were located in the above two domains. These results would be heplful for the design of HPV16 prophylactic peptide vaccines and HPV polyvalent vaccines.

Objective To systematically evaluate the risk factors associated with the occurrence of nosocomial infections(NI)in patients undergoing extracorporeal membrane oxygenation(ECMO)support.Methods A computerized systematic search was performed aross the Chinese databases including CNKI,Wanfang Database,China Biomedical Literature Database(CBMdisc),and Weipu,as well as the English databases such as PubMed,EMBase,Web of Science,and the Cochrane Library for case-control or cohort studies on the risk factors of hospital-acquired infections in patients undergoing ECMO support from the time of database construction to February 2023.The relevant literatures were screened by two researchers independently.Meta-analysis was performed using RevMan 5.4 software.Results A total of 20 papers,including 2 746 patients and 16 risk factors,were included.Meta-analysis results showed that older age[>50 years:odds ratio(OR)= 2.87,95％ confidence interval(95％ CI)was 1.24-6.63,P = 0.01;>65 years:OR = 1.66,95％ CI was 1.22-2.26,P = 0.001],combined hypertension(OR = 1.48,95％ CI was 1.01-2.15,P = 0.04),combined diabetes mellitus(OR = 1.40,95％ CI was 1.02-1.94,P = 0.04),sequential organ failure assessment(SOFA)was higher(OR = 1.06,95％ CI was 1.02-1.10,P = 0.000 7),the installation of ECMO in the intensive care unit(ICU,OR=1.48,95％ Ciwas 1.11-1.99,P=0.008),ECMO course(OR=1.27,95％ Ciwas 1.05-1.54,P = 0.01),ventilator-assistance for >48 hours(OR = 4.91,95％ CI was 2.40-10.05,P<0.000 1),and tracheotomy(OR = 9.56,95％ CI was 3.60-25.35,P<0.000 01)were identified as ECMO risk factors for hospital-acquired infections in patients.Conclusion Older age,combined hypertension,diabetes mellitus,higher SOFA,ECMO installation site in ICU,ECMO course,ventilator assistance>48 hours,tracheotomy are the risk factors for the occurrence of hospital-acquired infections in patients with ECMO,healthcare professionals should promptly identify the risk factors related to hospital-acquired infections,and take active and effective measures against controllable risk factors,including early intervention to prevent the occurrence of NI in ECMO patients.

OBJECTIVETo assess the association of polymorphisms of human leukocyte antigen (HLA)-A, -B, -DRB1 alleles and haplotypes with acute lymphoblastic leukemia (ALL) among ethnic Hans from northern China.

METHODSA total of 170 ALL patients (patient group) and 1241 unrelated healthy bone marrow donors (control group) were genotyped at a high-resolution level using polymerase chain reaction-sequence-based typing (PCR-SBT), sequence specific oligonucleotide probes (SSO) and sequence specific primer (SSP) typing methods. Frequencies of HLA alleles and haplotypes were calculated with Arlequin 3.5.2 software. The distribution of genes and haplotypes were analyzed through a case-control study, and the odd ratio (OR) of ALL was also calculated.

RESULTSBy cha-square test and correction, an increased frequency of B*13:01 and B*40:02 among ALL patients was discovered in comparison with the controls (7.35% vs. 4.63%, P=0.030; 2.94% vs. 1.45%, P=0.042), whereas B*35:03 and B*46:01 were less frequent compared with the controls (0.29% vs. 1.69%, P=0.048; 4.41% vs. 7.82%, P=0.025). Although the above discrepancies were not statistically significant by Bonferroni correction, within DRB1*15 group, the frequency of DRB1*15:01 in ALL patients was significantly greater than that of the controls (16.18% vs. 10.19%, Pc'=0.041) and was correlated with ALL (OR=1.70, 95% CI:1.24-2.33). Nineteen haplotypes identified in the ALL patients had a frequency greater than those of the controls. Of these, 11 were absent from the control group and were correlated with ALL.

CONCLUSIONThe association of HLA-A, -B, -DRB1 polymorphisms with ALL was determined among patients from northern Chinese Hans. The correlation between DRB1*15:01 and ALL suggested that DRB1*15:01 may be a susceptibility gene for ALL with its particular haplotypes.

Adolescent ; Adult ; Alleles ; Case-Control Studies ; Child ; Child, Preschool ; Female ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DRB1 Chains ; genetics ; Haplotypes ; Humans ; Infant ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Young Adult

OBJECTIVE: To verify a rare allele of human leukocyte antigen (HLA) and analyze its inheritance and 3D molecular structure. METHODS: PCR-sequence-based typing, PCR-single strand oligonucleotide polymorphism and single allele-specific sequencing were carried out to characterize the rare HLA-C allele and its transmission in the family. Its protein structure was modeled by using SWISS-MODEL, Phyre2 and FATCAT software. RESULTS: Analysis indicated that the rare allele (HLA-C*08:84) has transmitted from the proband's mother and has differed from HLA-C*08:01 by a single base (g.512G>C), resulting in substitution of an amino acid (p.Trp147Ser). Modeling of the 3D structure of the encoded protein indicated that the amino acid residue variation is located at the alpha 2 helix, which participates the formation of pocket F. Modeling of the structures of C*08:84, C*08:01, C*08:02, C*08:03 and C*08:22 has suggested significant variation in the peptide binding regions of the backbone, with root mean square errors being 1.70 nm, 1.79 nm, 0.71 nm and 1.70 nm, respectively. CONCLUSION A rare HLA-C*08:84 allele has been identified, and its clinical significance has been analyzed.
Alleles ; Base Sequence ; HLA-B Antigens/genetics* ; HLA-C Antigens/genetics* ; Humans ; Molecular Structure ; Sequence Analysis, DNA

Objective:To analyze a Chinese pedigree with a recombination occurring between the HLA- A/ C loci in both parents. Methods:A patient who was planning to undergo hematopoietic stem cell transplantation due to "aplastic anemia" in February 2022 was selected as the study subject. Peripheral blood samples were collected from the patient, his parents and brother. HLA- A/ C/ B/ DRB1/ DQB1 high-resolution typing was carried out by using sequence-based typing and sequence-specific oligonucleotides. The recombination was identified by pedigree analysis. The HLA haplotype of each individual was identified by genealogical analysis. The parentage possibility was determined by short tandem repeat analysis. HLA- A/ C/ B/ DRB1/ DRB345/ DQA1/ DQB1/ DPA1/ DPB1 were determined with next-generation high-throughput sequence-based typing. The recombination sites were analyzed by family study. Results:The high parentage possibilities of the family was confirmed by short tandem repeat analysis. Recombination was found between the HLA- A* 24: 02 A* 33: 03/ C* 14: 03 in the paternally transmitted haplotype, whilst HLA- A* 01: 01 A* 03: 01/ C* 08: 02 was found in the maternally transmitted haplotype, which had resulted in two novel HLA haplotypes in the proband. Conclusion:A rare case with simultaneous recombination of the paternal and maternal HLA- A/ C loci has been discovered, which may facilitate further study of the mechanisms of the HLA recombination.

【Objective】 To evaluate the appropriate optimal capacity and matching probability of the platelet donor database with known HLA/HPA genotype in Shaanxi aera, and provide data support for subsequent construction, maintenance and application of the local platelet donor database. 【Methods】 A total of 11 755 individuals from the Shaanxi Branch of China Marrow Donor Program, 401 and 249 unrelated random platelet donors in Shaanxi aera were enrolled to the population study of HLA-A, -B polymorphisms, HPA genotyping and CD36 antigen expression, respectively. The frequencies of HLA-A, -B alleles, HPA alleles and haplotypes were calculated with Arlequin 3. 5. 2. 2 software; matching probability and capacity evaluation of platelet donor database was conducted according to the phenotypic frequencies. 【Results】 The population genetic and phenotypic polymorphisms data of HLA-A, -B and HPA1-6, 10, 15, 21 in Shaanxi aera were obtained. The frequency of CD36 type Ⅰ or Ⅱ deficiency was 0.40%(1/249). According to the subsequent calculating and deriving, with a database size of 194 donors, the patient having approximate 95% probability could achieve matching of HPA1-6, 10, 15, 21 genotype. With a database size of 1500 donors, there is a 95% probability of matching at least one donor with HLA-A-B phenotype frequency >0.002 or haplotype frequency >0.001; meanwhile, the probability of matching a cross-reactive group donor should be 44.95%-97.57%. Based on database size of 8 856 and 15 033, the probabilities of matching HLA-A, -B phenotype were about 80% and 90%, respectively. 【Conclusion】 The differences in the distribution of HLA/HPA polymorphism in different regions make the establishment mode and optimal capacity of platelet donor database different. It is necessary to apply a variety of platelet matching transfusion strategies to expand the range of donor selection, thereby effectively reducing the database construction cost and resource requirements.