Objective:To explore the clinical and genetic characteristics of a fetus diagnosed with Congenital myasthenic syndrome type 16 (CMS16).Methods:A couple who had visited Tianjin Medical University General Hospital in February 2018 due to "adverse outcome of two pregnancies" was selected as the study subject. Clinical data was gathered. Peripheral blood and amniotic fluid samples were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. Low-depth whole-genome sequencing was carried out to detect copy number variation (CNV) in the fetus.Results:The couple′s first pregnancy had resulted in a miscarriage at 27 + 5 weeks, when ultrasound had revealed pleural effusion and polyhydramnios in the fetus. Their second pregnancy was terminated at 30 + 5 weeks due to fetal hand malformations, polyhydramnios and pleural fluid. Both couple had denied family history of genetic conditions. For their third pregnancy, no CNV abnormality was detected, whilst a compound heterozygous variants, including a maternally derived c. 3172C>T (p.R1058W) and paternal c. 1431delG (p.K477fs*89) in the SCN4A gene were detected. Based on the guidelines from the American College of Medical Genetics and Genomics, the c. 3172C>T (p.R1058W) was predicted as a likely pathogenic variant (PM1+ PM2_supporting+ PP3+ PP4), whilst the c. 1431delG (p.K477fs*89) was predicted as a pathogenic variant (PVS1+ PM2_supporting+ PP4). Conclusion:The c. 3172C>T (p.R1058W) and c. 1431delG (p.K477fs*89) compound heterozygous variants of the SCN4A gene probably underlay the CMS16 in the third fetus.

Objective:To explore the clinical and molecular basis for a Chinese pedigree affected with Complete androgen insensitivity syndrome (CAIS).Methods:A CAIS pedigree presented at Tianjin Medical University General Hospital between 2019 and 2021 was selected as the study subject. Clinical data of the proband was collected, along with peripheral blood samples from the proband and her family members. Chromosomal karyotyping, sex-determining region of the Y chromosome ( SRY) testing, and next-generation sequencing (NGS) were carried out for the proband, and candidate variant was verified by Sanger sequencing of her family members. Prenatal diagnosis was provided for the sister of the proband. This study was approved by Medical Ethics Committee of the Tianjin Medical University General Hospital (Ethics No. IRB2023-WZ-070). Results:The 18-year-old proband, who has a social gender of female, underwent laparoscopic examination, which showed no presence of uterus and ovaries. The karyotype of peripheral blood sample was 46, XY, with SRY gene detected. NGS indicated that the proband has harbored a heterozygous c. 1988C>G (p.Ser663Ter) variant of the AR gene. Sanger sequencing confirmed that her mother and sister had both harbored the same variant, whilst her father and younger sister were of the wild-type. Prenatal diagnosis revealed that her sister′s first fetus had harbored carried the same variant, which had led to termination of pregnancy. Her second fetus did not carry the variant, and a healthy boy was born. Based on guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as likely pathogenic (PM2_Supporting+ PM4+ PP3_Moderate+ PP4). Conclusion:The c. 1988C>G (p.Ser663Ter) variant of the AR gene probably underlay the CAIS in the proband. The accurate diagnosis of sex development disorders will rely on the physicians′ thorough understanding of the clinical symptoms and pathogenic genes. Genetic testing and counseling can enable precise diagnosis, prenatal diagnosis, and guidance for reproduction

Objective:To analyze the clinical and laboratory characteristics of acute myeloid leukemia (AML) patients with NPM1 mutation, and to explore the prognostic factors.Methods:A total of 77 AML patients with NPM1 gene mutation admitted to Hebei Yanda Ludaopei Hospital from May 1st 2012 to December 31st 2021 were enrolled in the study, including 34 male and 43 female patients. The median age was 40 (3, 68) years old. Patients were selected and divided into 4 groups according to the morphological FAB classification. There were 29 cases (37.7%) of M1 type, 13 cases (16.9%) of M2 type, 23 cases (29.9%) of M4 type, and 12 cases (15.5%) of M5 type. The clinical characteristics, bone marrow/peripheral blood cell morphology, immunophenotype, cytogenetics, molecular biology and overall survival of different groups were retrospectively analyzed, and the risk factors affecting the prognosis of AML were also explored. Cox multivariate regression was used to analyze the clinical influencing factors of survival and prognosis.Results:The white blood cell counts were highest in M4 and M5 patients and lowest in M2 patients, while no significant difference in the red blood cell, hemoglobin, and platelet counts( P>0.05). Morphologically, there were significant differences in the percentage of blasts and blasts with cup-like nuclei on bone marrow (BM) and peripheral blood (PB). The proportion of blasts in BM and PB was the highest in M1 and the lowest in M2 ( P<0.001). The positive rate of blasts with cup-like nuclei was the highest in M1 and the lowest in M5 of BM ( P<0.001), while the highest in M2 and the lowest in M5 of PB ( P=0.006). The scores of myeloperoxidase and chloroacetate esterase were all the highest in M1 and the lowest in M5 ( P<0.001, 0.001, respectively). In terms of molecular biology, the occurence rate of blasts combined with DNMT3A mutation was the highest in M4 and the lowest in M2 ( P=0.044), while those combined with FLT3-ITD mutation was the highest in M4 and the lowest in M5 ( P=0.002). In immunophenotype, there were significant differences in the expression positivities of seven antigens including HLA-DR, CD56, CD11c, CD15, CD14, CD96 and cMPO ( P<0.05). Multivariate COX regression analysis showed that no recurrence after treatment ( P<0.001), complete remission after treatment ( P=0.015) and transplantation ( P<0.001) were correlated with overall survival (OS). No recurrence after treatment ( P=0.033), transplantation ( P=0.027), no mutation of FLT3-ITD ( P=0.040), and hemoglobin concentration ( P=0.023) were associated with relapse-free survival (RFS). Survival analysis by Kaplan-Meier curve showed that there was no significant difference in survival time between the M1, M2, M4 and M5 groups in OS and RFS. Conclusion:There were significant differences in the white blood count, the percentage of blasts and blasts with cup-like nuclear morphology, cytochemical staining (MPO integration, CE integration and percentage of NAS-DCE), gene mutation (DNMT3A and FLT3-ITD) and immunophenotypes (HLA-DR, CD56, CD11c, CD15, CD14, CD96 and cMPO) between the four groups. The multivariate analysis revealed that no recurrence after treatment and transplantation were independent prognostic factors in NPM1 mut AML patients. On the other hand, FLT3-ITD mutation and hemoglobin concentration were associated with RFS and complete remission after treatment was associated with OS in the entire NPM1 mut cohort.

Objective:To investigate the correlation between hematopoietic cell kinase (HCK) and the expression level of the mitochondrial ribosomal protein L13 (MRPL13) and hematopoietic multimode ultrasound.Methods:204 female breast cancer patients treated by surgery in Quzhou people’s Hospital from Jan. 2017 to Sep. 2020 were selected as study subjects. Breast cancer tissues and adjacent normal tissues were extracted intraoperatively. Preoperative conventional ultrasound, shear wave elastography (SWE) and contrast-enhanced Ultrasonography (CEUS) were used to detect HCK and MRPL13 expression levels. Univariate analysis and binary Logistic regression were used to analyze the correlation between multi-mode ultrasonic features and HCK and MRPL13.Results:The positive expression ratios of HCK and MRPL 13 in breast cancer tissues were significantly higher than those in adjacent tissues ( χ2 was 5.625, 7.197; P was 0.018, 0.007) . In conventional ultrasound features, the proportions of HCK-positive breast cancer patients with irregular mass edges, microcalcifications, and grade II to III blood flow classification were significantly higher than those of HCK-negative patients ( χ2 was 7.437, 16.684, 23.262; P was 0.006, <0.001, <0.001) ; The proportion of MRPL13-positive breast cancer patients with a maximum diameter of ≥2 cm, irregular edges of the tumor, and grade II-III blood flow classification was significantly higher than that of MRPL13-negative patients ( χ2 was 4.676, 11.118, 8.389; P was 0.031, 0.001, 0.004) . For SWE signs, the proportion of HCK positive breast cancer patients with hard ring sign was significantly higher than that of HCK negative patients ( χ2=11.220, P=0.001) ; the proportion of MRPL13 positive breast cancer patients with hard ring sign and black hole sign was significantly higher than that of MRPL13. Those who were negative ( χ2 was 4.482, 8.775; P was 0.034, 0.003) . Among CEUS characteristics, the proportion of HCK-positive patients with high enhancement was significantly higher than that of HCK-negative patients ( χ2=7.356, P=0.007) ; the proportion of MRPL13-positive patients with high enhancement and late regression was significantly higher than that of MRPL13-negative patients ( χ2 was 9.165, 7.631; P was 0.002, 0.006) . The results of binary logistic analysis showed that there was microcalcification ( OR=4.619, 95% CI=2.657-8.119, P=0.009) , blood flow classification II to III ( OR=4.150, 95% CI=2.547-7.954, P=0.015) and high enhancement of CEUS ( OR=4.150, 95% CI=2.547-7.954, P=0.015) are independent risk factors for positive expression of HCK; blood flow grade II to grade III ( OR=4.213, 95% CI=3.145-8.557, P=0.012) , appearance of black hole sign ( OR=5.246, 95% CI=2.864-10.378, P<0.001) and high enhancement of CEUS ( OR=3.872, 95% CI=1.887~6.438, P=0.026) were the independent risk factors for the positive expression of MRPL13. Conclusion:The multimodal ultrasonographic features of breast cancer are helpful to predict the expression levels of HCK and MRPL13, so as to provide new imaging ideas for early diagnosis of breast cancer, the designation of treatment options and the preoperative non-invasive assessment of breast cancer prognosis.

Objective:To understand the quality of life of high risk population of stroke in community and analyze the influencing factors.Methods:Four community health service centers in Huangpu District were randomly selected, and the subjects were included by using convenient sampling method among the high risk groups of stroke found in the community stroke screening and prevention and control project in Shanghai.The World Health Organization quality of life scale-brief form questionnaire(WHOQOL-BREF), self-rating depression scale (SDS) and self-rating anxiety scale (SAS) were used as the survey tools to study 1200 high-risk stroke subjects.Single sample t-test was used to compare the differences between the scores and the data of 24 centers in the world, and multiple linear stepwise regression were used to analyze the influencing factors. Results:The scores of physical field, psychological field and social relationship field ((13.60±1.86), (14.58±1.97), (13.72±2.10)respectively) of high-risk population of stroke were lower than the scores of general population of 24 centers in the world ( P<0.01), and the scores of environmental field (14.08±1.95) were higher than it ( P<0.01), of which the differences were all statistically significant.The results of multiple linear stepwise regression analysis showed that old age, anxiety and depression were the risk factors influencing the scores of physical field( β=-0.027, -0.056, -0.051), psychological field( β=-0.019, -0.055, -0.050) and environmental field( β=-0.017, -0.040, -0.054); old age and depression were the risk factors influencing the scores of social relationship field( β=-0.026, -0.067); anxiety and depression were the risk factors influencing the self-assessment of quality of life and health ( β=-0.012, -0.014 for quality of life; -0.012, -0.014 for health, all P<0.01). Conclusion:The quality of life of high-risk population of stroke may be related to age, depression, anxiety and other psychological factors.The prevention and treatment of stroke should take both physical and mental measures, take timely intervention for poor psychological status, and gradually improve the quality of life.

OBJECTIVETo analyze clinical manifestations and genetic mutation in a child with severe short stature and other malformations.

METHODSThe child has undergone history taking and physical examination. Genome DNA was extracted from peripheral blood samples of the proband and her family members. Candidate genes were captured with Agilent SureSelect and sequenced on an Illumina platform. Suspected mutation was verified by Sanger sequencing.

RESULTSThe patient, a six-year-and-10-month old girl, presented with non-symmetrical short stature, dysmorphism, abnormalities of limbs and spine, amblyopia of left eye, and cataract of right eye, in addition with frequent respiratory infection and micturition. Laboratory testing suggested 25-hydroxy vitamin D deficiency (18.9 ng/mL). Spine X-ray showed multiple malformations with centrums. Her mother also featured short stature (138 cm). Her aunt had short stature (130 cm) and limb-length discrepancy. Her little brother was 2.5 years old, and his height was 81 cm (-3.4 SD). Exome sequencing revealed a heterozygous mutation c.184C to T (p.Arg62Trp) in the proband and her mother. The same mutation was not found in her father and brother.

CONCLUSIONThe patient was diagnosed with X-linked chondrodysplasia punctata 2. Mutation of the EBP gene probably underlied the disease in this family.

Objective: To establish an accurate and selective UPLC-MS/MS) method for the determination of afatinib in rat plas-ma. Methods: Protein precipitating by acetonitrile was used to prepare the samples. A CORTECS BEH C18column ( 50 mm × 2. 1 mm, 1. 6 μm) was used to separate the analytes at 40℃. The mobile phase consisted of acetonitrile and water (0. 1% formic acid) with the flow rate of 0. 4 ml·min-1. The analytes were quantified by multiple reaction monitoring ( MRM) mode with positive electrospray ionization, while the target fragment ions were m/z 486. 19→112. 1 for afatinib and m/z 557. 3→112. 15 for neratinib (IS). Results: The calibration curve obtained good linearity for afatinib within the range of 1–200 ng·ml-1(r=0. 998 1), and the LLOQ in rat plasma was 1. 0 ng/ml. The intra-and inter-day precisions were both≤9. 51% . The recovery of afatinib from plasma was above 77. 1% . After intragastric administration and intravenous administration of afatinib in rats, the t1/2was 7. 19 h and 2. 69 h, Cmax was 97. 78 ng·ml-1and 123. 37 ng·ml-1,and AUC(0-∞)was 1 505. 4 ng·ml-1·h and 405. 55 ng·ml-1·h, respectively. Con-clusion: The validated method can be applied in the pharmacokinetic study of afatinib at the intragastric and intravenous dosage of 10 and 2 mg·kg-1, respectively.

Objective To study the clinical and pathological features of histiocytic necmtizing lym-phadenitis( HNL) in children. Methods The clinical data and histological findings of 38 cases of HNL admitted in our hospital from June 2000 to May 2015 were reviewed. Results Most of the patients were school-age children with male-femal ratio of 1. 4: 1. The main clinical features were lymphadenopathy (100%),fever(68. 24%),leucocytopenia(52. 63%),rising of lymphocytes percentage(84. 21%). All of the lymph node excisional biopsy met the criterion of HNL. Some cases spontaneously relieved and some cases were treated with NSAID,glucocorticoid or immunoglobulin and benefited significantly. There was no recurrence. Conclusion The clinical situation is not specific. The diagnosis is established by lymph node ex-cisional biopsy. HNL is benign and self-limited disease. The effect of management using glucocorticoid, NSAID and immunoglobulin is remarkable. Long term follow-up is necessary.

Objective To investigate the characteristics and essential points of diagnosis and treatment of double trisomy 47,XXX/48,XXX,+8 combined Behcet disease, a rare inherited immunodeficiency disorder. Methods The clinical manifestations, karyotype analysis and gene test results of the patients were analyzed, and relevant literatures were reviewed. Results A 11-year-old girl presented repeated fever for more than 6 years, accompanied with recurrent genital herpes infection and oral apthosis, was clinically diagnosed with Behcet disease. Cytogentic and molecular karyotyping on peripheral lymphocytes demonstrated 47,XXX[12]/48,XXX,+8[18]. Conclusions Conventional karyotype analysis and chromosomal microarray analysis have a complementary role in the diagnosis of the disease. We conclude that patients with constitutional trisomy 8 and those with trisomy 8 confined to the bone marrow are both at increased risk of developing features of Behcet disease. The mechanism may relate to increased gene dosage of candidate genes for Behcet's disease on chromosome 8.

To implement the excellent doctor education training plan and construct the training model of practical ability in clinical education,the researchers analyzed the problems existing in the training of clinical practice ability and put forward a new train of thought,and constructed a training method system of personal experience,reading classics,teacher guidance,simulation training,clinical practice and reflection strengthening.In addition,we also established a safeguard mechanism with the characteristics of the organizational corporation of teaching,the expert-oriented teaching team,team-oriented mentoring,pro-ject-oriented ability training,the diversification of evaluation system,the modernization of teaching platform and the realistic training environment,to train the qualified clinical medical talents.