Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Objective To investigate the effects of Epigallocatechin gallate(EGCG)on IL-1βinduced MIN6 cells apoptosis. M.Methods The experiment group was divided into control group,IL-1β group,IL-1β+ EGCG low concentration group and IL-1β+EGCG high concentration group.Cell activity was detected by CCK8.Insulin secretion was detected by ELISA.cell apoptosis was detected by flow cytometry.The mitochondrial membrane potential was detected by flow cytometry.ATP content and cell ac-tivity of ROS were detected by colorimetry and chemiluminescence method.Results Compared with normal group,IL-1β group showed much lower cell activity,insulin secretion,cell mitochondrial membrane potential and ATP content,and at the same time IL-1βgroup had significantly higher cell apoptosis and ROS activities.After given EGCG,both low concentration group and high con-centration group had higher cell activity,insulin secretion,cell mitochondrial membrane potential and ATP content,at the same time lower cell apoptosis and ROS activities was showed.And the IL-1β+EGCG high concentration group worked more powerful.Con-clusion EGCG has protective effects on IL-1βinduced MIN6 cells apoptosis.Its mechanism may be related to increasing the content of the ATP and mitochondrial membrane potential and protecting mitochondrial function as well reducing the activity of ROS.

Objective To systematically evaluate the association between vascular endothelial grow th factor‐C (VEGF‐C) ex‐pression in breast cancer tissue and clinical significance in the domestic patients with breast cancer by a Meta‐analysis .Methods The published case controlled trials on the VEGF‐C expression and the clinical manifestations of breast cancer were retrieved from the CNKI ,CBM ,VIP and Wanfang databases ,and other relevant journals were also manually retrieved to identify all the relevant case controlled trials .The retrieval year limit was from the database establishment to June 2014 .The included literatures were screened according to the inclusion and exclusion standards and the quality of included case controlled trials was assessed .The Rev‐Man 5 .2 software was used to conduct the Meta analysis .Results A total of 15 case controlled trials involving 975 patients with breast cancer were included .The Meta analysis results revealed that there were statistical differences in the VEGF‐C expression be‐tween the breast cancer group and the control group[OR = 8 .16 ,95% CI(5 .77 ,11 .54)] ,between the lymph node metastasis posi‐tive group and the non‐lymph node metastasis negative group[OR = 5 .19 ,95% CI(3 .63 ,7 .44)] and between the clinical stage Ⅰ -Ⅱ group and the stage Ⅲ - Ⅳ group[OR = 0 .35 ,95% CI(0 .21 ,0 .59)] ;the difference in the VEGF‐C expression between the 0 - <50 years group and the ≥ 50 years group had no statistical significance ,indicating that the VEGF‐C expression had no obvious as‐sociation with the patient′s age .Conclusion The present evidences reveal that VEGF‐C maybe participate in the development and progression process of lymph node metastasis of breast cancer and may be become an important factor influencing the prognosis of breast cancer .