1.Construction of the core competence evaluation index system for emergency medical dispatchers
Qiuxia ZHAO ; Xuezhang ZHANG ; Yanqiu HAO ; Yufang GAO ; Ziya XIAO
Chinese Journal of Modern Nursing 2024;30(16):2143-2150
Objective:To construct an evaluation index system for the core competence of emergency medical dispatchers, so as to provide a theoretical basis for the training, assessment and ability evaluation of emergency medical dispatchers.Methods:In September 2022, a research group was established to compile the first draft of the evaluation index system for the core competencies of emergency medical dispatchers through literature review, semi-structured interviews and group discussions. From October to November 2022, a total of 27 experts in related fields from nine provinces and municipalities directly under the Central Government in China were consulted by Delphi expert letters, items were screened and amended, and an evaluation index system for the core competence of emergency medical dispatchers was formed.Results:A total of two rounds of correspondence were conducted. The effective recovery rate of the questionnaire was 100%, and the authority coefficient of the experts was 0.856. Kendall's coordination coefficients were 0.402 and 0.414 ( P<0.01), respectively. Finally, a core competency evaluation index system for emergency medical dispatchers was constructed, which included seven primary indicators, 16 secondary indicators and 75 tertiary indicators. Conclusions:The core competence evaluation index system for emergency medical dispatchers is scientific, reliable and comprehensive, which can provide a theoretical basis for the training, assessment and ability evaluation of emergency medical dispatchers.
2.Current status and influencing factors of intrinsic capacity in elderly patients with hospitalization-associated disability
Mengya HAN ; Xinyu ZHAO ; Yanqiu WANG ; Ludan XU ; Yuhua LIU ; Binru HAN
Chinese Journal of Modern Nursing 2024;30(19):2533-2538
Objective:To explore the current status of intrinsic capacity in elderly patients with hospitalization-associated disability (HAD) and explore its influencing factors.Methods:From November 2023 to January 2024, convenience sampling was used to select 203 elderly patients with HAD at Xuanwu Hospital of Capital Medical University as the study subjects. A survey was conducted on elderly patients using the General Information Questionnaire, Fried Frailty Phenotype, Barthel Index, Social Support Rating Scale, and Intrinsic Capacity Assessment Tool. Binomial Logistic regression was used to analyze the influencing factors of intrinsic capacity in elderly patients with HAD.Results:A total of 203 questionnaires were distributed, and 199 valid questionnaires were collected, with a valid response rate of 98.03% (199/203). The total score of intrinsic capacity in 199 elderly patients with HAD was 5.00 (4.00, 6.00), with scores for cognitive dimension, psychological dimension, motor dimension, vitality dimension, and sensory dimension being 1.00 (1.00, 2.00), 2.00 (1.00, 2.00), 0 (0, 1.00), 1.00 (1.00, 1.00) and 1.00 (1.00, 1.00), respectively. The binomial Logistic regression showed that department of medicine and surgery, self-rating health status, social support, serum albumin, and Barthel Index were the influencing factors of intrinsic capacity in elderly patients with HAD ( P<0.05) . Conclusions:The intrinsic capacity of elderly patients with HAD is at medium to low level, with the most severe impairment in the motor dimension. Medical and nursing staff should develop personalized rehabilitation measures for elderly HAD patients based on the influencing factors of their intrinsic capacity, enhance their intrinsic capacity, and reduce the burden of care on families and society.
3.Analysis of the status and influencing factors of different types of cognitive frailty in aged inpatients with hypertension
Shuao TANG ; Mengyao WEI ; Ludan XU ; Xinyu ZHAO ; Binru HAN ; Yanqiu WANG
Chinese Journal of Modern Nursing 2024;30(26):3566-3573
Objective:To investigate the status and influencing factors of different types of cognitive frailty in aged inpatients with hypertension.Methods:Totally 300 aged patients with hypertension admitted to the Cardiology, Geriatrics, and Endocrinology Departments of Xuanwu Hospital, Capital Medical University, from April to October 2022 were selected by convenience sampling. Data were collected using a general information questionnaire, the Fried Frailty Phenotype (FP), the Mini-Mental State Examination (MMSE), the 9-item Subjective Cognitive Decline Questionnaire-9 (SCD-Q9), the Morisky Medication Adherence Scale (MMAS), the Pittsburgh Sleep Quality Index (PSQI), the Hospital Anxiety and Depression Scale (HADS), the Social Support Rating Scale (SSRS), and the Mini Nutritional Assessment (MNA) .Results:A total of 300 questionnaires were distributed and returned, yielding a 100.00% response rate. Among the 300 aged hypertensive patients, 74 were diagnosed with reversible cognitive frailty (RCF) and 98 with potentially reversible cognitive frailty (PRCF). Logistic vegression analysis showed that, factors influencing RCF included daily exercise habits and the use of thiazide diuretics ( P<0.05) ; factors influencing PRCF included age, pre-retirement occupation, daily exercise habits, sleep quality, Barthel Index score, and serum albumin levels ( P<0.05) . Conclusions:The status of both RCF and PRCF is relatively high among aged inpatients with hypertension. The influencing factors for different types of cognitive frailty vary. Targeted interventions based on both common and specific influencing factors should be developed to reverse or slow the progression of cognitive frailty in aged hypertensive patients.
4.Preliminary study of ABI3BP-knockout mouse simulating low birth weight model
Yanqiu HUANG ; Yue ZHANG ; Liuliu SHI ; Xiaoying ZHAO ; Junming TANG ; Yan WU
Acta Laboratorium Animalis Scientia Sinica 2024;32(10):1307-1312
Objective To employ a mouse model of ABI3BP gene deletion for the detection of postnatal changes in body weight and glucose metabolism and establish a different method of creating a mouse model of low birth weight.Methods Heterozygote mice were mated to produce ABI3BP gene knockout homozygote(ABI3BP-/-)mice,heterozygote(ABI3BP+/)mice,and wild-type(WT)mice.Adult mice from all three groups were evaluated for glucose metabolism markers,including the fasting blood glucose level,glucose tolerance,and insulin tolerance.Additionally,body weight was measured at various postnatal time periods,and the weight ratio of critical organs in adulthood was calculated.Results The gene sequencing result of the polymerase chain reaction product of ABI3BP-/-mice showed that frameshift mutations occurred in the knockout region,with quantitative reverse-transcription polymerase chain reaction analysis demonstrating significantly reduced ABI3BP expression in ABI3BP-/-mice compared with that in WT mice.Notably,the birth weight of ABI3BP-/-mice(1.25±0.08 g)was markedly lower than that of WT mice(1.34±0.12 g)(P<0.05).Conversely,the weight of adult(120 d)ABI3BP-/mice(27.70±1.93 g)was significantly higher than that of WT mice(23.64±1.34 g)(P<0.01).The ratios of key organ weights to body weight were not significantly different between the groups(P>0.05).Fasting blood glucose and insulin tolerance tests showed no significant variations between the groups.However,glucose tolerance tests indicated that ABI3BP-/-mice had lower blood glucose levels(15.68±7.04 mmol/L)than WT mice(23.01±5.75 mmol/L).Conclusions Deletion of the ABI3BP gene result in mice with low birth weight,poor growth recuperation,and inadequate glucose tolerance in adulthood,similar to the clinical growth traits of low-birth-weight human neonates.Therefore,this mouse model is a promising choice for the study of low birth weight.
5.Effects of preventative moxibustion at Shenque(CV8)and Guanyuan(CV4)on BDNF-TrkB pathway in the spinal dorsal horn and hypothalamus in rats with dysmenorrhea
Xiangyun MENG ; Yulei LIANG ; Di WANG ; Xuan ZHANG ; Yanqiu SUN ; Yanxue XING ; Zhiguo ZHAO ; Xiaoshun MA ; Xinhua LI ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(3):195-203
Objective:To observe the effects of preventative moxibustion on analgesia,substance P(SP),prostaglandin(PG)F2α and PGE2 in rats with dysmenorrhea due to cold-dampness stagnation,and to explore the analgesic mechanism. Methods:Sixty-four female Wistar non-pregnant rats were randomly divided into a blank group,a model group,a Western medicine group,and a preventative moxibustion group,with 16 rats in each group.Eight qualified diestrus rats were selected from each group.Except for the blank group,the other three groups established models of dysmenorrhea due to cold-dampness stagnation using an ice water bath combined with estradiol benzoate and oxytocin.On the 8th day after modeling,the preventative moxibustion group was treated with gentle moxibustion at Shenque(CV8)and Guanyuan(CV4),and the Western medicine group was given ibuprofen solution for 4 consecutive days.On the 11th day,the intervention groups(i.e.the Western medicine group and the preventative moxibustion group)were treated once again after being injected with oxytocin.The writhing score and the pain threshold of rats were determined;the serum levels of brain-derived neurotrophic factor(BDNF),SP,PGF2α,and PGE2 were measured;the mRNA and protein expression levels of BDNF and its receptor tropomyosin receptor kinase B(TrkB)in the spinal dorsal horn and hypothalamus were detected. Results:Compared with the blank group,the writhing score increased(P<0.01),the pain threshold decreased(P<0.01),the serum levels of BDNF,SP,and PGF2α increased(P<0.01),while the PGE2 decreased(P<0.01);the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus increased(P<0.01)in the model group.Compared with the model group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α levels decreased significantly,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus decreased significantly in the preventative moxibustion group and the Western medicine group,while the inter-group differences were significant(P<0.01).Compared with the Western medicine group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α,levels decreased,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus in the preventative moxibustion group decreased significantly,while the inter-group differences were significant(P<0.05 or P<0.01). Conclusion:Preventative moxibustion at Shenque(CV8)and Guanyuan(CV4)can improve the pain sensitization state of rats with dysmenorrhea due to cold-dampness stagnation,down-regulate the mRNA and protein expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus;regulation of the serum SP,PGF2α,and PGE2 levels may be part of the mechanism.
6.Treatment status of tyrosine kinase inhibitor for newly-diagnosed chronic myeloid leukemia: a domestic multi-centre retrospective real-world study
Xiaoshuai ZHANG ; Bingcheng LIU ; Xin DU ; Yanli ZHANG ; Na XU ; Xiaoli LIU ; Weiming LI ; Hai LIN ; Rong LIANG ; Chunyan CHEN ; Jian HUANG ; Yunfan YANG ; Huanling ZHU ; Ling PAN ; Xiaodong WANG ; Guohui LI ; Zhuogang LIU ; Yanqing ZHANG ; Zhenfang LIU ; Jianda HU ; Chunshui LIU ; Fei LI ; Wei YANG ; Li MENG ; Yanqiu HAN ; Li'e LIN ; Zhenyu ZHAO ; Chuanqing TU ; Caifeng ZHENG ; Yanliang BAI ; Zeping ZHOU ; Suning CHEN ; Huiying QIU ; Lijie YANG ; Xiuli SUN ; Hui SUN ; Li ZHOU ; Zelin LIU ; Danyu WANG ; Jianxin GUO ; Liping PANG ; Qingshu ZENG ; Xiaohui SUO ; Weihua ZHANG ; Yuanjun ZHENG ; Qian JIANG
Chinese Journal of Hematology 2024;45(3):215-224
Objective:To retrospectively analyze the treatment status of tyrosine kinase inhibitors (TKI) in newly diagnosed patients with chronic myeloid leukemia (CML) in China.Methods:Data of chronic phase (CP) and accelerated phase (AP) CML patients diagnosed from January 2006 to December 2022 from 77 centers, ≥18 years old, and receiving initial imatinib, nilotinib, dasatinib or flumatinib-therapy within 6 months after diagnosis in China with complete data were retrospectively interrogated. The choice of initial TKI, current TKI medications, treatment switch and reasons, treatment responses and outcomes as well as the variables associated with them were analyzed.Results:6 893 patients in CP ( n=6 453, 93.6%) or AP ( n=440, 6.4%) receiving initial imatinib ( n=4 906, 71.2%), nilotinib ( n=1 157, 16.8%), dasatinib ( n=298, 4.3%) or flumatinib ( n=532, 7.2%) -therapy. With the median follow-up of 43 ( IQR 22-75) months, 1 581 (22.9%) patients switched TKI due to resistance ( n=1 055, 15.3%), intolerance ( n=248, 3.6%), pursuit of better efficacy ( n=168, 2.4%), economic or other reasons ( n=110, 1.6%). The frequency of switching TKI in AP patients was significantly-higher than that in CP patients (44.1% vs 21.5%, P<0.001), and more AP patients switched TKI due to resistance than CP patients (75.3% vs 66.1%, P=0.011). Multi-variable analyses showed that male, lower HGB concentration and ELTS intermediate/high-risk cohort were associated with lower cytogenetic and molecular responses rate and poor outcomes in CP patients; higher WBC count and initial the second-generation TKI treatment, the higher response rates; Ph + ACA at diagnosis, poor PFS. However, Sokal intermediate/high-risk cohort was only significantly-associated with lower CCyR and MMR rates and the poor PFS. Lower HGB concentration and larger spleen size were significantly-associated with the lower cytogenetic and molecular response rates in AP patients; initial the second-generation TKI treatment, the higher treatment response rates; lower PLT count, higher blasts and Ph + ACA, poorer TFS; Ph + ACA, poorer OS. Conclusion:At present, the vast majority of newly-diagnosed CML-CP or AP patients could benefit from TKI treatment in the long term with the good treatment responses and survival outcomes.
7.Predictive value of lactate/albumin ratio,interleukin-6 and CD4+T lymphocyte count in the short-term prognosis of severe pneumonia and sepsis
Zhiyu MIAO ; Lei ZHANG ; Xiaoyan LI ; Ruixia ZHANG ; Mengjiao ZHAO ; Yanqiu GAO
Journal of Xinxiang Medical College 2024;41(3):232-239
Objective To investigate the predictive value of lactate/albumin ratio(LAR),interleukin-6(IL-6)and CD4+T lymphocyte count in 28-day mortality in patients with severe pneumonia and sepsis.Methods A total of 73 patients with severe pneumonia and sepsis admitted to the Respiratory Intensive Care Unit(RICU)of Zhengzhou Central Hospital Affiliated to Zhengzhou University from January 2022 to June 2023 were enrolled and divided into the survival group(n=43)and the death group(n=30)according to their 28-day outcomes.The clinical data of the patients were collected from their electronic medical records,including age,gender,comorbidities with hypertension,diabetes,and coronary artery heart disease(CHD),as well as sequential organ failure assessment(SOFA)score,acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score,mean arterial pressure(MAP),confusion,uremia,respiratory rate,blood pressure,age ≥65 years(CURB-65)score,total bilirubin(Tbil),serum creatinine(Scr),platelet count(PLT),white blood cell(WBC)count,procalcitonin(PCT),and C-reactive protein(CRP)at admission to RICU.On the 1st,3rd,and 7th day after admission to RICU,the patients'arterial blood was drawn,and the lactate level was detected by a fully automated blood gas analyzer.The peripheral venous blood was drawn,and the serum albumin and IL-6 levels were detected by enzyme-linked immunosorbent assay,and the CD4+T lymphocyte subset count was measured by flow cytometry.The LAR of patients on the 1st,3rd and 7th day was calculated.The clinical data of the patients and the LAR,IL-6 level and CD4+T lymphocyte count on the 1st,3rd,and 7th day were compared between the two groups.The influencing factors of 28-day mortality in patients with severe pneumonia and sepsis were analyzed by logistic regression,and the predictive value of each influencing factor on the 28-day mortality in patients with severe pneumonia and sepsis was evaluated by the receiver operating characteristic(ROC)curve.Results There was no significant difference in gender,age,proportions of comorbidities with hypertension,diabetes and CHD,length of stay in RICU,and Tbil,MAP,PLT,Scr,WBC,PCT and CRP at admission to RICU(P>0.05).The APACHE Ⅱ and CURB-65 scores of the patients in the death group were significantly higher than those in the survival group(P<0.05).On the 1st,3rd and 7th day,the CD4+T lymphocyte count in the death group was significantly lower than that in the survival group,while the SOFA score was significantly higher than that in the survival group(P<0.05).On the first day,there was no significant difference in the LAR and IL-6 level be-tween the death group and the survival group(P>0.05).However,on the 3rd and 7th day,the LAR and IL-6 level in the death group were significantly higher than those in the survival group(P<0.05).The LAR,IL-6 level and SOFA score on the 3rd and 7th day in the survival group were significantly lower than those on the 1st day,and these indicators on the 7th day were sig-nificantly lower than those on the 3rd day(P<0.05);the CD4+T lymphocyte count on the 3rd and 7th day was significantly higher than that on the 1st day(P<0.05),while it showed no significant difference on the 7th and 3r day(P>0.05).The IL-6 level on the 7th day in the death group was significantly lower than that on the 1st and 3rd day(P<0.05),while there was no significant difference in IL-6 level on the 1st day compared with the 3r day(P>0.05);moreover,there was no significant difference in LAR,CD4+T lymphocyte count and SOFA score between each time point(P>0.05).Pearson correlation analy-sis showed that on the 3rd day,the LAR and IL-6 level were significantly positively correlated with the SOFA score in patients with severe pneumonia and sepsis(r=0.385,0.394;P<0.05).On the 7th day,the LAR and IL-6 level were also significantly positively correlated with the SOFA score(r=0.418,0.402;P<0.05).On the 3 rd and 7 th day,CD4+T lymphocyte count was significantly negatively correlated with the SOFA score(r=-0.451,-0.454;P<0.05).Logistic regression analysis showed that the APACHE Ⅱ score,LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day,and the IL-6 level and CD4+T lym-phocyte count on the 7th day were the influencing factors for 28-day mortality in patients with severe pneumonia and sepsis(P<0.05).The ROC curve showed that the APACHE Ⅱ score,LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day and the combination of the three,IL-6 level and CD4+T lymphocyte count on the 7th day and the combination of the two had certain predictive value for the 28-day mortality in patients with severe pneumonia and sepsis(P<0.05).The area under the ROC curve(AUC)of LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day combined to predict 28-day mortality in patients with severe pneumonia and sepsis was 0.891,and the AUC of APACHE Ⅱ score for predicting 28-day mortality in pa-tients with severe pneumonia and sepsis was 0.769.The AUC values of LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day for predicting 28-day mortality in patients with severe pneumonia and sepsis were 0.795,0.757 and 0.770,respective-ly,and the AUC values of IL-6 level and CD4+T lymphocyte count on the 7th day and their combination for predicting 28-day mortality in patients with severe pneumonia and sepsis were 0.743,0.802 and 0.888,respectively.Conclusion The 3-day LAR,IL-6 level and CD4+T lymphocyte count,and the 7-day IL-6 level and CD4+T lymphocyte count after admission are re-lated to the 28-day mortality in patients with severe pneumonia and sepsis.The combined LAR,IL-6 level and CD4+T lympho-cyte count on the 3rd day can better assess the severity and prognosis of patients.
8.Effects of Mild Moxibustion on Proteins Expression of BDNF/TrkB Signaling Pathway in Hypothalamus of Primary Dysmenorrhea Rats with Cold and Dampness Stagnation Syndrome
Xiangyun MENG ; Fei GAO ; Di WANG ; Yulei LIANG ; Xuan ZHANG ; Yanqiu SUN ; Yanxue XING ; Zhiguo ZHAO ; Xinhua LI
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(3):127-131
Objective To observe the analgesic effects of mild moxibustion on primary dysmenorrhea(PD)rats with cold and dampness stagnation syndrome and its effect on BDNF/TrkB signaling pathway in hypothalamus;To explore its mechanism for the treatment of PD.Methods A total of 32 Wistar non-pregnant female rats were randomly divided into blank group,model group,Western medicine group and mild moxibustion group,with 8 rats in each group.Except for the blank group,the other groups received estradiol benzoate intraperitoneal injection combined with ice bath treatment + oxytocin intraperitoneal injection to establish PD with cold and dampness stagnation syndrome model.The mild moxibustion group received treatment at"Shenque"and"Guanyuan"from the eighth day of modeling for 10 min,and the Western medicine group was given ibuprofen solution intragastically for 4 days.The latency period of rats twisting was observed and the twisting score was calculated,Western blot and PCR were used to detect the expressions of c-fos,BDNF,TrkB protein and mRNA in hypothalamic tissue.Results Compared with the blank group,the model group showed a shortened latency period and an increased twisting score(P<0.01),the expressions of c-fos,BDNF,TrkB protein and mRNA in hypothalamic tissue increased(P<0.01,P<0.05).Compared with the model group,the mild moxibustion group had a longer latency period and lower twisting score(P<0.01),while the expressions of c-fos,BDNF,TrkB protein and mRNA in hypothalamic tissue increased(P<0.01,P<0.05).Conclusion Mild moxibustion may effectively improve the pain state of PD rats with cold and dampness stagnation syndrome.This mechanism may be related to downregulating c-fos expression,inhibiting BDNF/TrkB signaling pathway activation,thereby inhibiting pain signal transmission,regulating pain occurrence and maintenance.
9.Mechanism of Wumeiwan in Treatment of Emotional Disorders Based on Secrets for Auxiliary Cultivation Life
Huimin YUAN ; Meilin ZHU ; Dan ZHAO ; Xiao CHEN ; Yanqiu DING ; Lijun MAO
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(7):230-236
Wumeiwan are recorded in the Jueyin chapter of Treatise on Cold Damage and Miscellaneous Diseases (《伤寒杂病论》), and many doctors in the past dynasties believed that they had the effects of calming intestinal ascarid by warming Zang organs. The clinical application was limited. Since KE Qin, a doctor in the Qing dynasty, proposed that Wumeiwan were the primary prescription against Jueyin diseases, Wumeiwan have been valued by doctors, and their clinical application has also been expanded. The syndrome indicated for Wumeiwan is attributed, not only to the ascarid internal harassment, but to the abnormal Qi movement of wind and wood in Jueyin and the dysfunction of Yin and Yang. According to the functions of six meridians based on the theory of "open-close axis" in Huangdi's Internal Classic (《黄帝内经》) and the original texts of Treatise on Cold Damage and Miscellaneous Diseases, Yinshu refers to Jueyin. Wumeiwan, the primary prescription against Jueyin diseases, have the function of connecting Yin and Yang, and they can treat diseases caused by Qi disturbance of Jueyin and cold and heat in complexity. Sharing the same resource, the Secrets for Auxiliary Cultivation Life: The Essential Method of Using Herbal Medicine for the Differential Treatment of the Five Zang Organs (《辅行诀五脏用药法要》) and Treatise on Cold Damage and Miscellaneous Diseases both consult Models of the Classic of Decoction (《汤液经方》). The formulation rules of tonic/purgative and collapse-saving prescriptions in Secrets for Auxiliary Cultivation Life follow the recorded "Tang Ye Jing Fa Tu" (an image revealing the compatibility principles of Chinese herbal drugs) and are closely related to the five elements theory. By exploring the formulation rules in "Tang Ye Jing Fa Tu" and the five elements of drugs, this study analyzed Wumeiwan and proved that Wumeiwan mainly acted on the liver, spleen, and heart. According to the Five Zang-organs Storing Spirit theory from Huangdi's Internal Classic, the above three organs are closely related to emotional adjustment. Therefore, the treatment of emotional disorders caused by qi disturbance of Jueyin and cold and heat in complexity with Wumeiwan is supported by evidence. At the end of this article, the reports on the treatment of emotional disorders with Wumeiwan in recent years were listed and reviewed. This article provides new ideas for the clinical application of Wumeiwan in the treatment of emotional disorders and expands the application range.
10.Application of single sperm sequencing for the preimplantation genetic testing of a Chinese family affected with Spinal muscular atrophy.
Jia CHEN ; Xingwu WU ; Ge CHEN ; Pengpeng MA ; Wan LU ; Zhihui HUANG ; Cailin XIN ; Yan ZHAO ; Qiongfang WU ; Yanqiu LIU
Chinese Journal of Medical Genetics 2023;40(2):148-154
OBJECTIVE:
To assess the value of single sperm sequencing in preimplantation genetic testing for monogenic disease (PGT-M).
METHODS:
A Chinese couple with two children whom had died of Spinal muscular atrophy (SMA) and attended the Jiangxi Provincial Maternal and Child Health Care Hospital in June 2020 was selected as the subject. Eleven single sperm samples were isolated by mechanical immobilization and subjected to whole genome amplification. Real-time PCR and Sanger sequencing were used to detect the SMN1 variants in the single sperm samples. Genomic DNA of the wife, her parents and the husband, as well as one single sperm sample harboring the SMN1 variant and two single sperm samples without the variant were used for the linkage analysis. Targeted capture and high-throughput sequencing were carried out to test 100 single nucleotide polymorphisms distributed within 2 Mb up- and downstream the variant site. The haplotypes linked with the SMN1 variants were determined by linkage analysis. Blastocyst embryos were harvested after fertilizing by intracytoplasmic sperm injection. Cells from the trophoblasts of each embryo were biopsied and subjected to whole genome amplification and targeted capture and high-throughput sequencing to determine their carrier status. Chromosomal aneuploidy of wild-type embryos was excluded. An euploid embryo of high quality was transferred. Amniotic fluid sample was taken at 18 weeks of gestation to confirm the status of the fetus.
RESULTS:
Genetic testing showed that the couple both had deletion of exons 7 ~ 8 of the SMN1 gene. The wife has inherited the deletion from her father, while the husband was de novo. The haplotypes of the husband were successfully constructed by single sperm sequencing. Preimplantation genetic testing has indicated that 5 embryos had harbored the heterozygous variant, 4 embryos were of the wild type, among which 3 were euploid. Prenatal diagnosis during the second trimester of pregnancy has confirmed that the fetus did not carry the deletion.
CONCLUSION
By single sperm sequencing and PGT-M, the birth of further affected child has been successfully avoided.
Humans
;
Pregnancy
;
Female
;
Child
;
Male
;
Preimplantation Diagnosis
;
East Asian People
;
Semen
;
Genetic Testing
;
Muscular Atrophy, Spinal/genetics*
;
Aneuploidy
;
Blastocyst/pathology*
;
High-Throughput Nucleotide Sequencing
;
Spermatozoa

Result Analysis
Print
Save
E-mail