OBJECTIVE: To identify pathogenic mutation in a pedigree affected with brachydactyly and obesity. METHODS: Peripheral blood sample was collected for extraction of genomic DNA. Exons capture combined with next generation sequencing (NGS) was carried out to identify potential mutation. Sanger sequencing was used to verify the results. RESULTS: NGS has identified a novel heterozygous missense mutation (c.125A>C, p.Gln42Pro) in the exon 1 of PTHLH gene. The result was verified by Sanger sequencing. The mutations was derived from his mother. His uncle and sister have also carried the same heterozygous mutation. CONCLUSION A novel mutation of the PTHLH gene has been identified in a pedigree affected with brachydactyly type E2 and obesity.
Brachydactyly ; complications ; DNA Mutational Analysis ; Humans ; Mutation ; Obesity ; complications ; Pedigree

［摘要］ 目的：探讨穿膜蛋白125（transmembrane protein125，TMEM125）在肺腺癌组织与A549 细胞中的表达，以及影响细胞 的增殖和侵袭能力的分子机制。方法：从癌症基因组图谱（the cancer genome atlas，TCGA）数据库收集肺腺癌数据包，下载临床 信息及基因表达谱数据。分析TMEM125 在肺腺癌组织中的表达及其与患者总生存期的相关性。构建TMEM125 过表达A549 细胞株，以CCK-8 法、细胞划痕实验检测TMEM125 过表达对肿瘤细胞的增殖和迁移能力的影响；流式细胞术检测TMEM125 过 表达对A549 细胞的细胞周期和凋亡的影响。WB检测TMEM125 过表达对下游NF-κB信号通路、凋亡蛋白的影响；免疫共沉淀 法（co-immunoprecipitation，Co-IP）检测TMEM125 与NF- κB 抑制因子结合Ras 样2（NF- κB inhibitor interacting Ras-like 2， NKIRAS2）的相互作用。利用TNFα（10 ng/ml）处理TMEM125 过表达A549 细胞，CKK-8、流式细胞术及WB检测其对细胞增殖、 凋亡以及NF-κB信号通路相关蛋白表达的影响。去甲基化试剂地西他滨处理A549 细胞，qPCR和WB检测TMEM125 基因和蛋 白的表达。结果：TMEM125 mRNA在肺腺癌组织中表达水平显著低于正常组织（P<0.001），启动子甲基化水平显著高于正常组 织（P<0.001），并且低、中表达患者总生存期显著低于高表达患者（P<0.001）。过表达TMEM125 抑制了A549 细胞的增殖和迁移 （P<0.01），增加细胞G2/M 期，促进细胞凋亡（P<0.01）；过表达TMEM125 与NKIRAS2 相互作用，显著抑制NF- κB 的活性 （P<0.01）；地西他滨处理A549 细胞可促进TMEM125 表达并且抑制细胞增殖（P<0.01）。结论：启动子高甲基化水平降低了 TMEM125 基因表达，导致其抑制NF-κB活性功能和抑制细胞增殖的作用下降，并且降低了细胞对地西他滨的敏感性。

Objective:To investigate the risk factors of proteinuria in patients with hypertension in Qinghai-Tibet Plateau.Methods:From March 2019 to June 2020, prospective design was used to collect data of Qinghai-Tibet Plateau hypertension patients who were eligible for continuous enrollment in the Department of Cardiovascular Medicine in Hospital of Chengdu Office of People's Government of Tibet Autonomous Region. Questionnaire survey, physical examination and blood pressure measurement were performed on the selected patients. Fasting venous blood samples were collected for liver function test, blood lipid test, blood glucose test, and hemoglobin test, etc. Three times of morning urine samples were taken on different days, and urine protein creatinine ratio (UACR) was measured, UACR < 30 mg/g was negative for urinary protein, and UACR≥30 mg/g was positive for urinary protein. At the same time, the selected patients were examined by carotid artery color ultrasound and heart color ultrasound. The risk factors of proteinuria were analyzed.Results:A total of 588 patients with hypertension met the inclusion criteria, including 472 patients (80.3%) who received antihypertensive drug therapy, 239 patients (40.6%) had antihypertensive treatment compliance, and 252 patients (42.9%) reached the standard blood pressure after theropy. Hypertension was associated with diabetes mellitus in 150 patients (25.5%), and urinary protein was positive in 126 patients (21.4%). In univariate analysis, ethnic composition, systolic blood pressure [(138.19 ± 19.65) vs (133.16 ± 18.45) mmHg, 1 mmHg = 0.133 kPa], diastolic blood pressure [(85.80 ± 13.51) vs (83.17 ± 12.19) mmHg], uric acid [(411.79 ± 101.54) vs (379.96 ± 102.18) μmol/L], hemoglobin [(152.86 ± 30.70) vs (143.49 ± 21.15) g/L], pulmonary artery trunk width [(21.76 ± 3.94) vs (20.98 ± 3.34) mm], and ventricular septal thickness [(9.90 ± 1.70) vs (9.47 ± 1.60) mm] in the positive group ( n = 126) were significantly higher than those in the negative group ( n = 462, P < 0.01 or < 0.05). In multivariate logistic regression analysis, increased systolic blood pressure [odds ratio ( OR) = 1.015, 95% confidence interval (95% CI): 1.005 - 1.026], uric acid ( OR = 1.003, 95% CI: 1.001 - 1.005), and pulmonary artery trunk width ( OR = 1.058, 95% CI: 1.001 - 1.118) were risk factors for proteinuria; Tibetans had a decreased risk of proteinuria compared with Han ( OR = 0.505, 95% CI: 0.317 - 0.805), but increased hemoglobin had an increased risk of proteinuria compared with normal hemoglobin ( OR = 1.890, 95% CI: 1.231 - 2.903). Conclusion:In patients with hypertension at high altitude, increased hemoglobin, systolic blood pressure, uric acid, pulmonary artery trunk width, and Han nationality are risk factors for proteinuria.

Objective:To explore the clinical characteristics and genetic variants of two children with 3-hydroxy-3-methylglutaryl-coenzyme A lyase deficiency (HMGCLD).Methods:Two children with HMGCLD diagnosed at Henan Provincial Children′s Hospital respectively in December 2019 and June 2022 were selected as the study subjects. Clinical data and results of laboratory testing were analyzed retrospectively.Results:Both children had manifested with repeated convulsions, severe hypoglycemia, metabolic acidosis and liver dysfunction. Blood amino acids and acylcarnitine analysis showed increased 3-hydroxy-isovalyl carnitine (C5OH) and 3-hydroxy-isovalyl carnitine/capryloyl carnitine ratio (C5OH/C8), and urinary organic acid analysis showed increased 3-hydroxyl-3-methyl glutaric acid, 3-methyl glutaric acid, 3-methyl glutacoic acid, 3-hydroxyisoglycine and 3-methylprotarylglycine. Child 1 was found to harbor homozygous c. 722C>T variants of the HMGCL gene, which was rated as uncertain significance(PM2_Supporting+ PP3). Child 2 was found to harbor homozygous c. 121C>T variants of the HMGCL gene, which was rated as pathogenic(PVS1+ PM2_Supporting+ PP4). Conclusion:Acute episode of HMGCLD is usually characterized by metabolic disorders such as hypoglycemia and metabolic acidosis, and elevated organic acids in urine may can facilitate the differential diagnosis, though definite diagnosis will rely on genetic testing.

Objective:The clinical and molecular genetic characteristics of 46, XY disorders of sex development caused by NR5A1 gene variants in 15 cases were analyzed to improve the understanding of this disease. Methods:The clinical data of children with NR5A1 gene variants diagnosed at the Children′s Hospital Affiliated to Zhengzhou University from March 2016 to December 2021 were retrospectively analyzed. Whole exome sequencing was performed to confirm the candidate sites, and Sanger sequencing was performed for validation. The patients were treated and followed up according to their disease characteristics. Results:At the initial diagnosis, 5 of the 15 cases were raised as females and 10 as males. The gonadal tissue was testis without residual Müllerian or ooticular structure, and all had various degrees of genital abnormalities. The average EMS masculinity score was 4.8 (1~9), including micropenis (100.0%), hypospadia (86.7%), unfused scrotum (46.7%), and abnormal testicular position (60.0%), in which the hypospadias was Ⅱ°~Ⅳ°. There was no skin pigmentation in 5 patients with growth retardation. Chromosomol karyotypes were 46, XY, adrenocorticotropin and cortisol levels were normal, electrolyte levels were normal, HCG stimulation test in 5 cases had normal response, 9 cases had low response. Anti-Müllerian hormone and statin B had decreased abnormally with age. A total of 14 NR5A1 variants were detected in the 15 children, most of which occurred in exon 4, of which 9 variant loci were not included in the HGMD database as of December 2022. Conclusion:The clinical phenotype of 46, XY abnormal sexual development caused by NR5A1 gene variants is extensive, with the external genitals showing varying degrees of insufficient masculinization. Adrenal involvement is rare.

Objective:To analyze clinical and genetic characteristics of congenital hypogonadotropic hypogonadism(CHH) in children.Methods:Clinical data of 0-18 year old CHH patients diagnosed in the Department of Endocrinology, Genetics and Metabolism of Children′s Hospital Affiliated to Zhengzhou University from January 1, 2016 to December 31, 2023 were retrospectively analyzed, including their hormone levels and genetic test results.Results:A total of 95 patients with CHH were included. Among them, 25 were diagnosed before the age of 3, 37 between the ages of 3-14, and 33 were over 14 years old at the time of first diagnosis. The primary manifestations were micropenis(95 cases, 100%) and cryptorchidism(46 cases, 48.5%). The incidence of cryptorchidism was the lowest in the group over 14 years of age. Hormonal analysis revealed that the peak levels of LH following statin B and GnRH stimulation, the peak levels of FSH after GnRH stimulation, and testosterone levels following hCG stimulation were the highest in the infant group. Genetic analysis identified 20 CHH-related genes in 61 out of 77 cases.Double-gene mutation accounted for 7.8%(6/77) and triple-gene mutation accounted for 3.9%(3/77). The most common mutations were FGFR1(18/77, 23.4%), CHD7(12/77, 15.6%), PROKR2(11/77, 14.3%) and ANOS1(6/77, 7.8%). The incidence of cryptorchidism in these four genotypes was 50%, 75%, 45.5% and 83.3%, respectively. The incidence of testicular dysfunction was 22.2%, 16.7%, 27.3%, and 16.7%, respectively, with no statistical significance.Conclusion:The primary manifestation of CHH is micropenis and cryptorchidism. In children with CHH, the incidence of testicular Leydig cell and Sertoli cell dysfunction increased with age in CHH children. FGFR1, CHD7, PROKR2 and ANOS1 were common variants of CHH.

Objective To explore the clinical features of critical cases of coronavirus disease 2019 (COVID-19). Methods The clinical data of nine patients who were diagnosed with critical COVID-19 in Hainan General Hospital from January 21, 2020 to February 6, 2020 were retrospectively analyzed. RT-PCR testing for 2019 novel coronavirus (2019-nCoV) was performed with multi-sites synchronize specimens including pharyngeal swab, blood, excrement, and urine. The serum levels of leucocyte, C-reactive protein, procalcitonin and lactic acid between the improved group (five cases) and the deteriorated group (four cases) were compared. The t test was used for comparison of normally distributed continuous data between groups. Results There were eight males (88.9%) and 1 female enrolled. The patients aged 28-77 years old, with an age of (52.9±18.0) years. By March 4, 2020, all five cases in improved group were cured and discharged, three cases in deteriorated group died and 1case remained in critical condition. All multi-sites specimens of patients in improved group turned negative in 2-4 weeks of illness onset, while those of cases in deteriorated group showed sustained viral nucleic acid positive (up to 48th day of illness onset). The white blood cell counts ((13.52±8.24)×10 9 /L vs (10.49±4.46) ×10 9 /L), C-reactive protein ((139.71±87.46) mg/L vs (78.60±55.40) mg/L) and procalcitonin ((2.32±4.03) ng/mL vs (0.28±0.58) ng/mL) , lactic acid ((3.70±4.14) mmol/L vs (2.33±0.53) mmol/L) in deteriorated group were all significantly higher than those in improved group ( t =2.908, 5.009, 4.391 and 2.942, respectively, all P <0.01). A rapid rise of serum IL-6 level up to 8 500 pg/mL was observed in one patient three days prior to death. Conclusion Among the patients with critical COVID-19, serum levels of inflammatory cytokines of the death cases are higher than those of improved and discharged cases.