OBJECTIVE: To explore the clinical characteristics and genetic factors in four patients with Disorder of sex development (DSD). METHODS: Four patients who visited Tianjin Medical University General Hospital between January 2023 and January 2024, presenting with short stature, abnormal external genitalia, or infertility as their chief complaints, were selected as the study subjects. Clinical data were collected, and peripheral or umbilical cord blood samples were obtained for karyotyping analysis and low-depth whole-genome sequencing (CNV-seq). Quantitative fluorescence PCR (QF-PCR) was used to detect the sex-determining region Y (SRY) gene and azoospermia factor (AZF) on the Y chromosome, while fluorescence in situ hybridization (FISH) was employed to determine the location of the SRY gene. Whole exome sequencing (WES) was performed for genetic testing, and Sanger sequencing was used for familial validation of the candidate variants. The study procedure and protocol were approved by the Medical Ethics Committee of Tianjin Medical University General Hospital (Ethics No.: IRB2024-WZ-006). RESULTS: Case 1 had a karyotype of 45,X[22]/46,XY[8], with CNV-seq indicating a mosaic deletion of 7.44 Mb (copy number = 0.2) at Yp11.31-p11.2, a mosaic deletion of 5.32 Mb (copy number = 0.3) at Yq11.1-q11.221, and a deletion of 10.26 Mb (copy number = 0) at Yq11.221-q11.23. Y chromosome microdeletion analysis showed SRY and AZFa (+), AZFb+c (-). Case 2 had a karyotype of 45,X[12]/46,X,del(X)(q26.3)[18], with CNV-seq indicating a mosaic deletion of 132.62 Mb (copy number = 1.4) at Xp22.33-q26.3 and a deletion of 19.62 Mb (copy number = 1) at Xq26.3-q28. Case 3 had a karyotype of 46,XX, with CNV-seq showing two copies of the X chromosome and no Y chromosome. Y chromosome microdeletion analysis showed SRY (+) and AZFa+b+c (-), and FISH confirmed a translocation of the SRY gene to the terminal end of the short arm of the X chromosome. Case 4 had a karyotype of 46,XY, with CNV-seq showing one copy each of the X and Y chromosomes. Y chromosome microdeletion analysis showed SRY(+) and AZFa+b+c (+), and WES revealed a c.1103del variant in the AR gene (maternal origin), which was classified as a pathogenic variant based on the guidelines from the American College of Medical Genetics and Genomics (ACMG) (PVS1+PP1+PM2_Supporting). CONCLUSION The combined application of multiple detection techniques such as chromosomal karyotyping analysis, CNV-seq, QF-PCR, and WES can identify the genetic etiology of DSD patients, providing a basis for clinical consultation and treatment plan formulation.
Humans ; Male ; Female ; Chromosomes, Human, Y/genetics* ; Disorders of Sex Development/genetics* ; Sex-Determining Region Y Protein/genetics* ; Karyotyping ; In Situ Hybridization, Fluorescence ; Exome Sequencing ; Adult ; Child

To report 7 Chinese children with characteristic hyperpigmented macules on the forehead and temples. Among the 7 cases, there were 2 males and 5 females, with the age at onset ranging from 9 to 24 months (12.43 ± 5.32 months), and the disease duration being 1 - 4 months (2.57 ± 1.27 months). Skin examination revealed that the children presented with varying numbers of irregular brown macules and patches scattered on their foreheads and temples, without obvious desquamation. Dermoscopic examination revealed multiple yellowish-brown patches with irregular borders, and linear vessels were observable in some skin lesions. A diagnosis of acquired facial hyperpigmented macules was made in these children. The children received no treatment. After 2 years of follow-up, hyperpigmented macules completely subsided in 2 cases and regressed to varying degrees in 4 cases, while 1 case exhibited no changes in the skin lesions. Considering the literature and the cases discussed in this article, it is hypothesized that acquired facial hyperpigmented macules in young children may represent an independent condition.

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 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 assess the clinical efficacy and health economic value of non-invasive prenatal testing (NIPT) for the prenatal screening of common fetal chromosomal aneuploidies. METHODS: 10 612 pregnant women from October 2017 to December 2019 presented at the antenatal screening clinic of the General Hospital of Tianjin Medical University were selected as the study subjects. Results of NIPT and invasive prenatal diagnosis and follow-up outcome for the 10 612 pregnant women were retrospectively analyzed and compared. Meanwhile, NIPT data for two periods were analyzed for assessing the health economic value of NIPT as the second- or first-tier screening strategy for the prenatal diagnosis of fetal trisomies 21, 18 and 13. RESULTS: The NIPT was successful in 10 528 (99.72%) subjects, with the sensitivity for fetal trisomies 21, 18 and 13 being 100%, 92.86% and 100%, and the positive predictive value (PPV) being 89.74%, 61.90% and 44.44%, respectively. The PPV of NIPT for sex chromosome aneuploidies was 34.21%. Except for one false negative case of trisomy 18, the negative predictive value for trisomy 21, trisomy 13 and other chromosomal abnormalities were 100%. For pregnant women with high risk by serological screening, advanced maternal age or abnormal ultrasound soft markers, NIPT has yielded a significantly increased high risk ratio. There was no statistical difference in the PPV of NIPT among pregnant women from each subgroup. NIPT would have higher health economic value as a second-tier screening until 2019, while compared to 2015 ~ 2017, its incremental cost-effectiveness ratio as a first-tier screening had declined clearly. CONCLUSION The screening efficacy of NIPT for trisomies 21, 18 and 13 for a mixed population is significantly better than conventional serological screening, but it is relatively low for sex chromosomal abnormalities. NIPT can also be recommended for populations with relatively high risks along with detailed pre- and post-test genetic counselling. From the perspective of health economics, except for open neural tube defects, it is possible for NIPT to replace the conventional serological screening in the future as its cost continues to decrease.
Pregnancy ; Female ; Humans ; Trisomy/genetics* ; Retrospective Studies ; Prenatal Diagnosis/methods* ; Down Syndrome/genetics* ; Aneuploidy ; Chromosome Aberrations ; Trisomy 18 Syndrome/genetics* ; Sex Chromosome Aberrations ; Fetus

Objective:To evaluate the clinical efficacy of Jiawei Zhixiao Decoction and western medicine comprehensive therapy in acute attack of bronchial asthma of heat asthma syndrome.Methods:Randomized controlled trial. A total of 80 patients with acute exacerbation of bronchial asthma and TCM pattern heat asthma, who treated in the respiratory department of our hospital from January 2021 to December 2021, were selected. Accordlty to random number table method the patients were divided into the treatment group and control group, with 40 in each group. The patients in the control group were given comprehensive treatment of Western Medicine (bronchodilator and glucocorticoid, etc.). On the basis of comprehensive treatment of Western medicine, the treatment group was combined with Jiawei Zhixiao Decoction. All patients received a 14-day treatment. Before and after treatment, the TCM symptom scores were recorded, FEV1 and forced expiratory volume in one second to forced vital capacity ratio (FEV1/FVC) were measured by pulmonary function meter, and peak expiratory flow (PEF) was measured by peak expiratory flow meter. Serum TNF-α and IL-8 levels were detected by ELISA, and procalcitonin (PCT) was detected by electrochemiluminescence. The disease grade, clinical efficacy and adverse reactions were also recorded.Results:The total effective rate was 95.0% (38/40) in the treatment group and 80.0% (32/40) in the control group, the difference between the two groups was statistically significant ( χ2=4.11, P=0.043). After treatment, the TCM symptom score of the treatment group was significantly lower than that of the control group ( t=7.91, P<0.01). FEV1 [(2.83±0.37) L vs. (2.38±0.32) L, t=6.77], FEV1/FVC [(85.37±9.36) % vs. (75.50±10.24) %, t=4.50], PEF [(4.84±0.82) L vs. (3.92±0.43) L, t=6.28] was significantly higher than that of control group ( P<0.01). Serum IL-8 [(80.59±10.28) ng/L vs. (87.15±8.25) ng/L, t=3.15], TNF-α [(43.18±4.08) ng/L vs. (51.78±7.58) ng/L, t=6.32], PCT [(0.84±0.35) μg/L vs. (0.41±0.12) μg/L, t=7.35] were significantly lower than those in control group ( P<0.01). The improvement of asthma grade was significantly better than that of control group ( Z=17.86, P<0.05). During the observation period, there were no serious adverse reactions in both groups, and the safety was high. Conclusion:Application of Jiawei Zhixiao Decoction and western medicine comprehensive therapy in acute attack of bronchial asthma of heat asthma syndrome can effectively improve the TCM symptoms and lung function, attenuate the inflammation response, and alleviate the severity of asthma.

OBJECTIVE: To carry out prenatal diagnosis for a fetus with mosaicism Yq deletion. METHODS: A fetus with high risk of sex chromosomes indicated by non-invasive prenatal testing (NIPT) at Tianjin Medical University General Hospital in July 2021 was selected as the study subject. Prenatal diagnosis of the fetus was performed with combined G-banded chromosomal karyotyping, fluorescence in situ hybridization (FISH), copy number variation sequencing (CNV-seq), real-time fluorescence PCR (QF-PCR), and ultrasound examination. RESULTS: Analysis of the amniocytes at 23 gestational weeks had yielded a 45,X karyotype. However, FISH had shown signals of Y chromosome. Re-examination by cordocentesis had shown a mosaicism of 46,X,+mar[33]/45,X[17]. FISH showed that 69% of the cells had contained Y chromosome signals. The result of CNV-seq was seq[19]del(Y)(q11.1q12)(mos) chrY: g.13200001_ 28820000del (mosaicism rate = 64%), which suggested mosaicism for a Yq deletion, which encompassed the azoospermia factor (AZF) region. Deletion of the AZF region was verified by QF-PCR. The fetal karyotype was ultimately determined as mos46,X,del(Y)(q11.1)[33]/45,X[17]. Although ultrasound examination had shown no abnormality in the fetus, the couple had opted to terminate the pregnancy, and the induced fetus had a normal male appearance. CONCLUSION The combined use of multiple techniques is beneficial for accurate and rapid prenatal diagnosis. For fetuses with mosaicism chromosomal abnormalities, it may be difficult to accurately predict the postnatal phenotype. It is therefore necessary to further explore their genotype-phenotype correlation in order to provide better guidance upon genetic counseling.
Female ; Pregnancy ; Male ; Humans ; Mosaicism ; DNA Copy Number Variations ; In Situ Hybridization, Fluorescence ; Y Chromosome ; Fetus

Cartilage has poor self-recovery because of its characteristics of no blood vessels and high extracellular matrix. In clinical treatment, physical therapy or drug therapy is usually used for mild cartilage defects, and surgical treatment is needed for severe ones. In recent years, cartilage tissue engineering technology provides a new way for the treatment of cartilage defects. Compared with the traditional surgical treatment, cartilage tissue engineering technology has the advantages of small wound and good recovery. The application of microcarrier technology in the design of tissue engineering scaffolds further expands the function of scaffolds and promotes cartilage regeneration. This review summarized the main preparation methods and development of microcarrier technology in recent years. Subsequently, the properties and specific application scenarios of microcarriers with different materials and functions were introduced according to the materials and functions of microcarriers used in cartilage repair. Based on our research on osteochondral integrated layered scaffolds, we proposed an idea of optimizing the performance of layered scaffolds through microcarriers, which is expected to prepare bionic scaffolds that are more suitable for the structural characteristics of natural cartilage.
Cartilage ; Extracellular Matrix/chemistry* ; Technology ; Tissue Engineering/methods* ; Tissue Scaffolds/chemistry*

Objective:To discuss the effect of pharmacist intervention on the use of antimicrobial agents in the clinical pathway of community-acquired pneumonia ( CAP) in our hospital to standardize the rational medication and promote the rational use of antimicro-bial agents. Methods:Totally 100 bacterial CAP patients in 2013 ( before the intervention) and 2014 ( after the intervention) in the pneumology department were studied. The antibacterial drug cost, total hospitalization cost, use intensity of antimicrobial drugs, hospi-talization time, therapeutic effects and so on in the two groups were observed during the treatment. Results: There were significant differences between the two groups in antibacterial drug cost, total hospitalization cost, use intensity of antimicrobial drugs, hospitaliza-tion time and so on, whereas there was no significant difference in the curative effect. Conclusion:After the pharmacist intervention, the application of antibiotics is more rational, the antibiotics use density and per capita cost are reduced, the hospitalization day is shortened and the value of pharmacists is also improved.

Objective:To explore the relationship between angiotensin converting enzyme (ACE) gene insertion/dele‐tion (I/D) polymorphism and familial essential hypertension (EH) .Methods :Fluorescent in situ hybridization stai‐ning DNA sequencing test was used to detect ACE gene I/D genotype frequency and allele frequency in 46 familial EH patients (familial EH group) ,64 EH patients without family history (non‐familial EH group) and 43 healthy people (healthy control group) .Results:In healthy control group ,non -familial EH group ,familial EH group ACE gene DD genotype frequency was 11.6% ,32.3% and 37.0% ;distribution frequency of D allele was 33.7% , 52.3% and 57.6% respectively .Compared with healthy control group ,there were significant rise in ACE gene DD genotype and D allele frequency in familial EH group and non -familial EH group (P<0.05 or <0.01) ,but there were no significant difference between familial EH group and non‐familial EH group (P>0.05) .Conclusion:ACE gene DD genotype and D allele may be genetic predisposing genes of patients with essential hypertension ,and there is no significant difference in genetic constituent ratio between familial EH patients and non‐familial EH patients .