RESUMO
Echinococcosis poses a significant concern in the fields of public health and veterinary care as it can be transmitted between animals and humans. The primary endemic subtypes are cystic echinococcosis (CE) and alveolar echinococcosis (AE), which result from infestation by Echinococcus granulosus and Echinococcus multilocularis, respectively. A prominent epidemic of echinococcosis greatly affects the Tibet Autonomous Region (TAR) in China. A new technique called the loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) test is introduced in this research to differentiate between E. granulosus and E. multilocularis using their repetitive genetic sequences. The test is characterized by its portable nature, simple operation, quick result production, high sensitivity, and low susceptibility to aerosol contamination. The LAMP-LFD method demonstrated an exceptional minimal detection limit, reaching levels as low as approximately 1 fg/µL (femtogram per microliter) of genomic DNA. The assay's specificity was assessed, and no cross-reactivity was seen. A total of 982 dog fecal samples were collected from 54 counties in the TAR region between July 2021 and June 2022. The established method underwent validation using a commercially available ELISA kit. The agreement rate between the LAMP-LFD and ELISA methods was 97.25%, with a sensitivity of 96.05% and a specificity of 97.35%. The assay described in this study improves specificity by using a double-labeled probe, and it reduces the risk of false-positive results caused by aerosol contamination through the use of a sealed device. This makes it a suitable choice for quickly and accurately identifying the two main types of Echinococcus in field settings.
RESUMO
Objective: To explore the correlation between different CYP21A2 pathogenic gene mutations and clinical phenotypes in Congenital adrenal hyperplasia (CAH) patients. Moreover, combined with the specific phenotypes of patients in the clinic, diagnosis and treatment suggestions should be made for CAH patients. Methods: In this study, a genetic status of a Chinese family in three generations of 21-hydroxylase deficiency was comprehensively presented, and the pathogenic genes in the family were found and traced in detail. We measured CYP21A2 gene in this family by Sanger sequencing and MLPA. The trophoblast cells of female proband's embryos were detected by PGT-M which used Copy-Number Variations of a Single Human Cell and high throughput sequencing. The CYP21A2 gene mutation site in each embryo were detected by Sanger sequencing, whole genome sequencing and single nucleotide polymorphism (SNP). Results: There are many related pathogenic genes of CAH in this family. The female proband showed a compound heterozygous mutation in the CYP21A2 gene, including a CYP21A1P/A2 fusion gene (CH-8) (classical phenotype) and a new mutation c.1034T > C (p. L354S) (unknown clinical significance). In the proband's family, a heterozygous gene mutation of c.1034T > C and a CYP21A1P/A2 fusion gene (CH-8) was carried by her father and mother, respectively. Meanwhile, the husband of the proband also has a genetic family with related disease. Both the husband and his father carried the CYP21A2 gene c.844G > T heterozygous mutation, while his mother had no related mutation in the CYP21A2 gene. Furthermore, PGTM gene detection was carried out on the four blastocysts of the proband's offspring through IVF. The results showed that embryos T1, T2 and T4 all carried CYP21A1P/A2 fusion gene (CH-8), as well as embryo T3 carried c.1034T > C heterozygous mutation of maternal origin. Conclusion: This case is a family report showing a complete genetic map of the proband and her family, describing the genetic process of different pathogenic genes in detail and clearly corresponding to the patient's different phenotypes. It is speculated that the pathogenesis of CAH is caused by different mutations in the CYP21A2 gene and their interactions, which may affect the different phenotypes of CAH patients.
RESUMO
As a specific predictor of ovarian reserve, serum anti-Müllerian hormone (AMH) has become an area of intense research interest in the field of assisted reproductive technology. We assessed the relationship between AMH levels and pregnancy outcomes in Chinese patients and investigate the influencing factors of cumulative live birth in patients with high AMH levels. A total of 1379 patients starting their IVF/ICSI cycle were divided into normal (Group A, 1.1-4.0 ng/ml, n = 639) and high (Group B, > 4.0 ng/ml, n = 740) groups by serum AMH levels. Live birth rate (LBR), cumulative live birth rate (CLBR) and cumulative clinical pregnancy rate (CCPR) were also investigated. Compared with Group A, Group B had a significantly higher CLBR (65.80% vs. 43.95%) and CCPR (76.77% vs. 57.14%), respectively. Binomial logistic regression analysis showed that age over 40 years, LH/FSH > 2.5, total Gn dose and Gn duration, and greater than 4000 ng/ml serum E2 levels on HCG day were significantly associated with CLBR in Group B. The AUC value of CLBR averaged 0.664 (ranging from 0.621 to 0.706) (p < 0.001). The patients with high AMH levels had higher CPR, higher LBR, and lower MR with no statistically significant differences, although there were significant improvements in CLBR. Advanced age (> 40 years) still impacted CLBR, even in women with good ovarian reserves. Consequently, it is still recommended that patients over 40 years old with high AMH levels actively receive IVF treatment if they seek to become pregnant. PCOS diagnoses did not influence the CLBR. In summary, this study showed that serum AMH levels could positively predict patient ovarian responses and further affect pregnancy outcomes.
