Objective: To analyze the HLA typing and STR loci chimerism in a patient with recurrent acute lymphoblastic leukemia after HLA-haploidentical hematopoietic stem cell transplantation. Methods: HLA typing was performed on peripheral blood, buccal swabs and saliva samples after transplantation using PCR-sequence-specific oligonucleotide probes (PCR-SSOP) and next-generation sequencing (NGS). Additionally, STR analysis was conducted on these samples using a 21-locus STR assay kit to detect STR loci. Results: The HLA typing and STR locus outcomes of the patient's peripheral blood and the second saliva sample post-transplantation were in full concordance with the test results of the donor (father), whereas the HLA typing and STR locus results derived from the buccal swabs and the first saliva sample indicated chimerism between the donor and the recipient. Conclusion: In the follow-up and monitoring after transplantation, apart from focusing on peripheral blood samples, it is recommended to regularly monitor HLA typing and STR loci chimerism in patients' buccal swabs and saliva samples to comprehensively evaluate the transplantation effect and recurrence risk.

Objective: To analyze the HLA typing and STR loci chimerism in a patient with recurrent acute lymphoblastic leukemia after HLA-haploidentical hematopoietic stem cell transplantation. Methods: HLA typing was performed on peripheral blood, buccal swabs and saliva samples after transplantation using PCR-sequence-specific oligonucleotide probes (PCR-SSOP) and next-generation sequencing (NGS). Additionally, STR analysis was conducted on these samples using a 21-locus STR assay kit to detect STR loci. Results: The HLA typing and STR locus outcomes of the patient's peripheral blood and the second saliva sample post-transplantation were in full concordance with the test results of the donor (father), whereas the HLA typing and STR locus results derived from the buccal swabs and the first saliva sample indicated chimerism between the donor and the recipient. Conclusion: In the follow-up and monitoring after transplantation, apart from focusing on peripheral blood samples, it is recommended to regularly monitor HLA typing and STR loci chimerism in patients' buccal swabs and saliva samples to comprehensively evaluate the transplantation effect and recurrence risk.

Objective:To delineate a deletional mutation of the HLA-B gene in a Chinese pedigree.Methods:A female patient with acute myeloid leukemia who had visited Liuzhou People′s Hospital in April 2022 was selected as the study subject. Routine human leukocyte antigen (HLA) was determined by using PCR-sequence specific oligonucleotide polymorphism (PCR-SSOP) and PCR-sequence-based typing (PCR-SBT) methods. Next generation sequencing (NGS) was used to validate the candidate variant in the HLA-B gene.Results:The PCR-SBT and SSOP results for the HLA-B locus were inconsistent for the patient and her daughter. The SSOP results of the two individuals were HLA-B*35: 01, 40: 02 and HLA-B*35: 01, 40: 01, respectively. However, the PCR-SBT results has indicated a mismatch with the nearest HLA-B*35: 01 at exon 4. NGS results showed that the HLA-B*35: 01 had a 9 bp deletion in the intron 5. The patient′s husband was HLA-B*40: 01, 58: 01, which was normal. Conclusion:The variant in intron 5 of the HLA-B gene in this pedigree has mapped to a primer-binding region for the SBT reagent, which has affected the accuracy of PCR-SBT results.

Objective:To observe any effect of long-distance application of Oral Reading for Language with Aphasia (ORLA) training on patients with post-stroke aphasia.Methods:A total of 42 stroke survivors with aphasia were randomly divided into an inpatient group, and two online groups, each of 14. All three groups had their routine rehabilitation treatment supplemented with ordinary multimodal language therapy and ORLA. The inpatient group completed the routine in the rehabilitation treatment room, while the online groups completed it at home using Tencent video conferencing software. The conventional multimodal language therapy was conducted once daily, 5 days a week for 4 weeks. For the inpatient group and online group 2 the daily session lasted 30 minutes, while for the online 1 groups the daily length was doubled. The ORLA therapy was also conducted once daily, 5 days a week for 4 weeks, for the inpatient group and online group 2 the daily session lasted 1h, while for the online 1 groups the daily length was 30min.The speech function, reading ability and life quality of the three groups were evaluated before and after the intervention using the Western Aphasia Battery, the Chinese Standard Aphasia Examination Scale and the Chinese version of the Stroke Aphasia Quality of Life Scale.Results:After treatment, the average aphasia quotient (AQ), reading, naming and quality of life scores in all three groups had improved significantly compared with those before treatment. And related language ability scores (such as retelling, fluency, information volume, listening comprehension, etc.) had also improved significantly in all three groups. However, the average AQ, reading, and oral fluency scores of the inpatient group and online group 2 were significantly higher than those of online group 1. Significant improvement was also observed in the reading aloud and life quality of all three groups, but the average improvement in reading aloud was significantly greater in the inpatient group and in online group 2 compared to online group 1. The average life quality of the online groups was significantly superior to that of the inpatient group.Conclusion:Medium- and high-intensity ORLA synchronous remote speech rehabilitation can significantly improve the speech ability, reading ability and life quality of aphasic stroke survivors.

Objective:To explore the improvement of diaphragm function after early off-bed mobility intervention in intensive care unit (ICU) patients undergoing mechanical ventilation.Methods:A randomized controlled trial was conducted. A total of 147 adult patients undergoing mechanical ventilation admitted to ICU of Affiliated Hospital of Zunyi Medical University from October 2019 to March 2022 were enrolled. The patients were divided into control group and observation group by convenient sampling. Except for the different intervention programs of early mobility, other treatment and nursing of the patients in the two groups were carried out according to ICU routine. Progressive early activities were performed in the control group, while early off-bed mobility was performed in the observation group. The changes of diaphragm thickness at the end of inspiratory (DTei), diaphragm thickness at the end of expiratory (DTee) and diaphragm thickening fraction (DTF) before and 24, 48, 72 and 96 hours of intervention, and the duration of mechanical ventilation, length of ICU stay and 24-hour re-intubation rate after intervention were compared between the two groups.Results:Among the 147 patients, there were 4 cases of detachment in the control group and 5 cases of detachment in the observation group. Finally, 138 patients were enrolled, 69 cases in the control group and 69 cases in the observation group. There was no significant difference in gender, age, diagnosis of ICU, sedatives, muscle strength, ventilator model, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and DTei, DTee, DTF before intervention between the two groups. The DTei, DTee and DTF in both groups were increased gradually with the extension of intervention time, especially in the observation group [DTei (cm) at 24, 48, 72 and 96 hours of intervention in the observation group were 0.247±0.014, 0.275±0.016, 0.300±0.013 and 0.329±0.013, while in the control group were 0.242±0.015, 0.258±0.013, 0.269±0.014, and 0.290±0.017, effect of time: F = 993.825, P = 0.000, effect of intervention: F = 82.304, P = 0.000, interaction effect between intervention and time: F = 84.457, P = 0.000; DTee (cm) of the observation group were 0.213±0.014, 0.227±0.013, 0.243±0.016, 0.264±0.010, while in the control group were 0.213±0.016, 0.218±0.013, 0.224±0.013, 0.234±0.014, effect of time: F = 385.552, P = 0.000, effect of intervention: F = 28.161, P = 0.000, interaction effect between intervention and time: F = 45.012, P = 0.000; DTF of the observation group were (15.98±4.23)%, (21.35±4.67)%, (24.09±4.44)% and (25.24±3.74)%, while in the control group were (14.17±4.66)%, (18.11±3.92)%, (20.22±4.19)% and (20.98±4.12)%, effect of time: F = 161.552, P = 0.000, effect of intervention: F = 49.224, P = 0.000, interaction effect between intervention and time: F = -4.507, P = 0.000]. The duration of mechanical ventilation and length of ICU stay in the observation group were significantly shorter than those in the control group [duration of mechanical ventilation (hours): 112.68±12.25 vs. 135.32±22.10, length of ICU stay (days): 7.84±1.78 vs. 10.23±2.43, both P < 0.01]. However, there was no significant difference in 24-hour re-intubation rate between the observation group and the control group (0% vs. 2.90%, P > 0.05). Conclusions:Both early off-bed mobility and progressive early activities can prevent diaphragm weakness in ICU patients undergoing mechanical ventilation, and the effect of early off-bed mobility is better. Early off-bed mobility can significantly shorten the duration of mechanical ventilation and length of ICU stay, and it is safe and feasible.

OBJECTIVE: To delineate the characteristics of a novel HLA-DQB1 allele identified during routine HLA matching in a leukemia family. METHODS: The mother and brother of the patient were subjected to PCR sequence-specific oligonucleotide probe (SSOP), PCR sequence-based typ1ing (SBT), as well as next-generation sequencing (NGS). RESULTS: PCR-SBT revealed that the patient's mother and brother's HLA-DQB1 sequences did not fully match with any known allele combination. NGS revealed that the novel allele has differed from the closest matched DQB1*03:02 with a T>G substitution at position 233 in exon 2, which resulted in substitution of Valine at codon 46 by Glycine. Pedigree analysis confirmed that the novel HLA-DQB1 allele was inherited from his mother. CONCLUSION A novel HLA-DQB1 allele has been identified through next generation sequencing and was officially named as HLA-DQB1*03:362 by the World Health Organization HLA Factor Nomenclature Committee.
Alleles ; Base Sequence ; HLA-DQ beta-Chains/genetics* ; Humans ; Male ; Nucleotides ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVE: To verify a HLA-DQB1*03:90N allele and method to improve the accuracy of HLA typing. METHODS: A total of 2265 hematopoietic stem cell donors from Shenzhen Branch of China Marrow Donor Program in 2018 were initially detected by a PCR sequence-specific oligonucleotide probe (SSOP) method. Among these, a rare HLA-DQB1 allele was identified by sequence-based tying (SBT) and Ion Torrent S5 next generation sequencing (NGS). RESULTS: The SSOP typing result suggested the HLA-DQB1 to be a rare allele, while an insertion and a deletion was suspected in its exon 2 by SBT, which were confirmed by NGS as DQB1*03:90N and DQB1*06:01, respectively. CONCLUSION Rare alleles suspected by the SSOP method should be verified by other methods to ensure the accuracy of HLA genotyping. Rare alleles formed by deletions can be detected by NGS with accuracy.

Objective To determine the reproductive physiology and blood physiological and biochemical characteristics of SPF Yorkshire and Landrace swine.Methods Ten reproductive physiology parameters,19 blood physiological parameters and 18 blood biochemical parameters in SPF Yorkshire and Landrace swine were measured using conventional methods and the differences between population,between age groups and between both sexes were analyzed.Results There were no significant differences(P>0.05) in reproductive physiology parameters and most blood physiological and biochemical parameters of the SPF Yorkshire and Landrace swine.A few of parameters,such as blood physiological indices GRAN,HGB,RDW,PLT,PCT,and blood biochemical indices ALKP,CHOL,TBIL,BUN,showed significant difference(P<0.05) between populations,between age groups and between both sexes,however,the values of difference were rather small,deviated from the normal range.Conclusion The physiological and biochemical characteristics of SPF Yorkshire and Landrace swine are basically stable and there is no significant difference compared with other laboratory miniature pigs.This study will provide valuable basic data for raising velvet yield,establishment of animal models and evaluating the genetic quality of closed colony.

Objective To investigate the miRNA expression profiles in rectal cancer tissues and their associations with clinical pathological stage, depth of tumor invasion, and lymph node metastasis, and to evaluate the potential of miRNA as diagnostic and prognostic markers of rectal cancer. Methods Human miRNA microarray was used to profile miRNA expression in rectal cancer tissues and matched adjacent normal tissues (n=71). The up-regulated miR-93-5p and down-regulated miR-27a-3p were screened out, and the top differentially expressed miRNA were validated by quantitative real-time polymerase chain reaction ( qRT-PCR) . The relationship between the expression of miRNA and clinical parameters was analyzed by ANOVA and Spearman correlation. Results The expression of miR-27a-3p was down-regulated in miRNA microarray, but was up-regulated in qRT-PCR analysis;the data were relatively discrete. The expression of miR-93-5p was up-regulated in both miRNA microarray and qRT-PCR analysis;the expression level of miR-93-5p in rectal cancer tissues was 3165 times that in adjacent normal tissues ( P=00058);the expression level was correlated with tumor volume ( P= 0004 ) , and was positively correlated with the level of carcinoembryonic antigen ( CEA) before treatment ( P=0001) and the number of lymph nodes metastases (rs=0534, P=0005). Conclusions There is a differential miRNA expression pattern between rectal cancer tissues and matched adjacent normal tissues. The miR-93-5p is highly up-regulated in rectal cancer tissues and may serve as a diagnostic and prognostic marker of rectal cancer.

Objective The aim of this study was to investigate the polymorphism of SLA class II genes in Canadian SPF Yorkshire and Landrace pigs.Methods Blood samples were obtained from 15 SPF Yorkshire and 22 Landrace pigs for isolation of peripheral blood mononuclear cells respectively, and the DQB1, DRB1 and DQA genes were amplified by PCR after reverse transcription.SLA class II genes were obtained by analyzing the direct and cloning result.The polymorphism of alleles was analyzed using the DNAsp 5.0 software.Results A total of 25 alleles were identified at three genes, including eight DQB1, ten DRB1 and seven DQA, and three alleles were submitted the complete sequences for the first time.The official allele names were assigned as SLA-DQB1*0212 (KU754590), SLA-DQB1*0203 (KU754591) and DRB1*06:07(KU754601) by the SLA Nomenclature Committee.Three novel DQA alleles were discovered.Five of the 15 amino acids, one of the 16 amino acids and 11 of the 19 amino acids, which bind processing antigens, showed well conserved among the alleles of DQB1, DRB1 and DQA genes in the SPF Yorkshire and Landrace pigs, respectively.Neighbor-joining tree showed that the three genes were divided into two clusters, respectively.There was a close relationship between SPF Yorkshire and Landrace pigs and foreign Yucatan miniature pigs, and it showed no obvious genetic distance with other pigs.Conclusions A total of 25 SLA class II alleles have been identified successfully in this study, and there are more abundant polymorphism for them.There is a widely distribution for SLA class II alleles identified in this study in other pig breeds.It is critical for the eventual future use of SPF Yorkshire and Landrace pigs as classical laboratory animal models.