OBJECTIVE: To investigate the genetic mechanism for adverse pregnancies due to submicroscopic chromosomal structural variants in two cases, and to provide a precise guidance for preimplantation genetic testing. METHODS: Two families who had visited Chengdu Women's and Children's Central Hospital for reproduction guidance due to recurrent miscarriages, adverse pregnancy history and abnormal genetic testing of the offspring in June and December 2023 were selected as the study subjects. Chromosomal karyotyping and optical genome mapping (OGM) were carried out on peripheral blood samples from the two couples, and preimplantation genetic testing for structural rearrangement (PGT-SR) were performed on the blastocyst trophoblasts. This study was approved by the Medical Ethics Committee of the Hospital (Ethic No.: 2023-23). RESULTS: No abnormality was found on the G-banded karyotyping analysis for both couples. The OGM results revealed that the female partner of couple 1 had a translocation between 4pter-p16.3 (3.99 Mb) and 11pter-p15.4 (2.66 Mb), whilst no abnormality was found in the male partner. Similarly, the male partner of couple 2 had a translocation between 19q13.43-qter (1.90 Mb) and 22q13.31-qter (3.34 Mb). No abnormality was found in the female partner of couple 2. Neither breakpoints nor the adjacent region had involved an OMIM gene, except the formation of a fusion gene ZIM2-AS1-Z82186.1 (Both genes are non-coding, and the fusion gene was deemed as variant of unknown significance). PGT-SR of 11 blastocysts derived from couple 1 revealed that one embryo was suitable for priority transfer, three embryos were suitable for transfer, one embryo was recommended for genetic counselling, and six embryos were unsuitable for the transfer. For couple 2, six blastocysts were tested, of which only one embryo was deemed suitable for transfer. CONCLUSION When genetic testing of offspring indicates copy number variations such as deletions, duplications or mosaicism, the high-resolution OGM technique can be selected to screen parents for submicroscopic chromosomal structural variations. The result can facilitate accurate assessment for the risk of recurrence in offspring, selection of suitable method for reproduction, and identifying targets for PGT.
Humans ; Female ; Pregnancy ; Adult ; Male ; Karyotyping ; Chromosome Aberrations ; Abortion, Habitual/genetics* ; Preimplantation Diagnosis ; Genetic Testing

Objective:To analyze the clinical features and pathogenic genes of a family with congenital cataracts.Methods:A pedigree analysis was performed.A Han Chinese family initially diagnosed with congenital cataracts at The Fourth Hospital of Shijiazhuang in March 2024 was enrolled.The proband and selected family members underwent detailed ophthalmic examinations.Potential cataract-associated genetic variants in the proband were identified using whole exome sequencing (WES). Sanger sequencing was employed to confirm the presence of these variants in the proband and other family members.The identified variants were analyzed in accordance with the guidelines of the American College of Medical Genetics and Genomics (ACMG). This study adhered to the Declaration of Helsinki.The research protocol was approved by the Ethics Committee of The Fourth Hospital of Shijiazhuang (No.20230074). Both the subjects and their guardians were informed of the study purpose and voluntarily signed the informed consent form.Results:The pedigree included four generations comprising 15 individuals, with three patients identified across the second, third, and fourth generations.These cases included two males and one female, specifically the proband, his mother, and his eldest son.The observed inheritance pattern aligned with an autosomal dominant mode, characterized by the clinical presentation of bilateral cataracts.WES identified a novel frameshift insertion variant c. 270_271insA in exon 2 of the CRYAB gene in the proband, resulting in a valine-to-serine substitution at amino acid position 91.This variant induced early termination of translation following the expression of two additional amino acids, loss of 84 amino acids (p.V91Sfs2) and the production of a functionally impaired protein.The Sanger sequencing validation results were consistent with the co-segregation.According to the ACMG classification criteria (PM2+ PP1+ PVS1), the variant was classified as likely pathogenic. Conclusions:The frameshift insertion variant c. 270_271insA (p.V91Sfs2) in exon 2 of the CRYAB gene is likely the pathogenic cause of congenital cataract in this family.This is the first report of this variant.

Objective:To analyze the clinical features and pathogenic genes of a family with congenital cataracts.Methods:A pedigree analysis was performed.A Han Chinese family initially diagnosed with congenital cataracts at The Fourth Hospital of Shijiazhuang in March 2024 was enrolled.The proband and selected family members underwent detailed ophthalmic examinations.Potential cataract-associated genetic variants in the proband were identified using whole exome sequencing (WES). Sanger sequencing was employed to confirm the presence of these variants in the proband and other family members.The identified variants were analyzed in accordance with the guidelines of the American College of Medical Genetics and Genomics (ACMG). This study adhered to the Declaration of Helsinki.The research protocol was approved by the Ethics Committee of The Fourth Hospital of Shijiazhuang (No.20230074). Both the subjects and their guardians were informed of the study purpose and voluntarily signed the informed consent form.Results:The pedigree included four generations comprising 15 individuals, with three patients identified across the second, third, and fourth generations.These cases included two males and one female, specifically the proband, his mother, and his eldest son.The observed inheritance pattern aligned with an autosomal dominant mode, characterized by the clinical presentation of bilateral cataracts.WES identified a novel frameshift insertion variant c. 270_271insA in exon 2 of the CRYAB gene in the proband, resulting in a valine-to-serine substitution at amino acid position 91.This variant induced early termination of translation following the expression of two additional amino acids, loss of 84 amino acids (p.V91Sfs2) and the production of a functionally impaired protein.The Sanger sequencing validation results were consistent with the co-segregation.According to the ACMG classification criteria (PM2+ PP1+ PVS1), the variant was classified as likely pathogenic. Conclusions:The frameshift insertion variant c. 270_271insA (p.V91Sfs2) in exon 2 of the CRYAB gene is likely the pathogenic cause of congenital cataract in this family.This is the first report of this variant.

Objective:To determine the total and age-specific cut-off values of total prostate specific antigen (tPSA) and the ratio of free PSA divided total PSA (fPSA/tPSA) for screening prostate cancer in China.Methods:Based on the Chinese Colorectal, Breast, Lung, Liver, and Stomach cancer Screening Trial (C-BLAST) and the Tianjin Common Cancer Case Cohort (TJ4C), males who were not diagnosed with any cancers at baseline since 2017 and received both tPSA and fPSA testes were selected. Based on Cox regression, the overall and age-specific (＜60, 60-＜70, and ≥70 years) accuracy and optimal cut-off values of tPSA and fPSA/tPSA ratio for screening prostate cancer were evaluated with time-dependent receiver operating characteristic curve (tdROC) and area under curve (AUC). Bootstrap resampling was used to internally validate the stability of the optimal cut-off value, and the PLCO study was used to externally validate the accuracy under different cut-off values.Results:A total of 5 180 participants were included in the study, and after a median follow-up of 1.48 years, a total of 332 prostate cancer patients were included. In the total population, the tdAUC of tPSA and fPSA/tPSA screening for prostate cancer were 0.852 and 0.748, respectively, with the optimal cut-off values of 5.08 ng/ml and 0.173, respectively. After age stratification, the age specific cut-off values of tPSA in the <60, 60-<70, and ≥70 age groups were 3.13, 4.82, and 11.54 ng/ml, respectively, while the age-specific cut-off values of fPSA/tPSA were 0.153, 0.135, and 0.130, respectively. Under the age-specific cut-off values, the sensitivities of tPSA screening for prostate cancer in males <60, 60-70, and ≥70 years old were 92.3%, 82.0%, and 77.6%, respectively, while the specificities were 84.7%, 81.3%, and 75.4%, respectively. The age-specific sensitivities of fPSA/tPSA for screening prostate cancer were 74.4%, 53.3%, and 55.9%, respectively, while the specificities were 83.8%, 83.7%, and 83.7%, respectively. Both bootstrap's internal validation and PLCO external validation provided similar results. The combination of tPSA and fPSA/tPSA could further improve the accuracy of screening.Conclusion:To improve the screening effects, it is recommended that age-specific cut-off values of tPSA and fPSA/tPSA should be used to screen for prostate cancer in the general risk population.

Objective:To determine the total and age-specific cut-off values of total prostate specific antigen (tPSA) and the ratio of free PSA divided total PSA (fPSA/tPSA) for screening prostate cancer in China.Methods:Based on the Chinese Colorectal, Breast, Lung, Liver, and Stomach cancer Screening Trial (C-BLAST) and the Tianjin Common Cancer Case Cohort (TJ4C), males who were not diagnosed with any cancers at baseline since 2017 and received both tPSA and fPSA testes were selected. Based on Cox regression, the overall and age-specific (＜60, 60-＜70, and ≥70 years) accuracy and optimal cut-off values of tPSA and fPSA/tPSA ratio for screening prostate cancer were evaluated with time-dependent receiver operating characteristic curve (tdROC) and area under curve (AUC). Bootstrap resampling was used to internally validate the stability of the optimal cut-off value, and the PLCO study was used to externally validate the accuracy under different cut-off values.Results:A total of 5 180 participants were included in the study, and after a median follow-up of 1.48 years, a total of 332 prostate cancer patients were included. In the total population, the tdAUC of tPSA and fPSA/tPSA screening for prostate cancer were 0.852 and 0.748, respectively, with the optimal cut-off values of 5.08 ng/ml and 0.173, respectively. After age stratification, the age specific cut-off values of tPSA in the <60, 60-<70, and ≥70 age groups were 3.13, 4.82, and 11.54 ng/ml, respectively, while the age-specific cut-off values of fPSA/tPSA were 0.153, 0.135, and 0.130, respectively. Under the age-specific cut-off values, the sensitivities of tPSA screening for prostate cancer in males <60, 60-70, and ≥70 years old were 92.3%, 82.0%, and 77.6%, respectively, while the specificities were 84.7%, 81.3%, and 75.4%, respectively. The age-specific sensitivities of fPSA/tPSA for screening prostate cancer were 74.4%, 53.3%, and 55.9%, respectively, while the specificities were 83.8%, 83.7%, and 83.7%, respectively. Both bootstrap's internal validation and PLCO external validation provided similar results. The combination of tPSA and fPSA/tPSA could further improve the accuracy of screening.Conclusion:To improve the screening effects, it is recommended that age-specific cut-off values of tPSA and fPSA/tPSA should be used to screen for prostate cancer in the general risk population.

Objective:To explore the incidence and mortality of digestive system cancers, and the trend of the disease burden attributed to different risk factors in population in China.Methods:Data were obtained from the GLOBOCAN 2020 and the Global Burden of Disease Study in 2019 databases and only the data from the Chinese population were included. Using Excel 2019 and R 4.2.1 software, indicators including age-standardized incidence rate (ASIR), age-standardized mortality rate (ASMR), age-standardized disability-adjusted life year (DALY) rate and its rate of change were used to illustrate the disease burden of digestive system cancers attributed to different factors and their trends.Results:In 2020, the ASIR of digestive system cancers in China was 83.00/100 000, and the ASMR was 63.80/100 000. The numbers of digestive system cancer cases and deaths increased with age, and more cases and deaths occurred in men than in women in all age groups. The age-standardized DALY rate of esophageal cancer, gastric cancer and liver cancers showed decreasing trends in China from 1990 to 2019 (rate of change: -45.26%, -46.87%, and -65.63%, respectively), whereas the age-standardized DALY rate of pancreatic cancer, colorectal cancer and gallbladder and biliary tract cancer showed increasing trends (rate of change: 67.61%, 30.52%, and 7.21%, respectively). The trend of the mortality rate was consistent with the DALY rate. Compared with the age-standardized DALY rate attributed to behavioral factors, the annual proportion of the age-standardized DALY rate attributed to metabolic factors to the total age-standardized DALY rate of esophageal cancer, liver cancer, pancreatic cancer, and colorectal cancer increased from 1990 to 2019. There was no significant change in the rank of age-standardized DALY rate of gastric cancer, liver cancer, pancreatic cancer, and gallbladder and biliary tract cancer attributed to different risk factors in China from 1990 to 2019, but the rank of certain attributed risk factors for the age-standardized DALY rate of esophageal cancer and colorectal cancer moved ahead (esophageal cancer: high BMI; colorectal cancer: low milk intake, and low whole-grain intake).Conclusions:The incidence and mortality of digestive system cancers was serious in China in 2020, and the annual proportion of the disease burden of digestive system cancers attributed to metabolic factors increased from 1990 to 2019. The rank of attributed risk factors for several digestive system cancers changed significantly.

Tetrandrine(TET),a natural bisbenzyl isoquinoline alkaloid extracted from Stephania tetrandra S.Moore,has diverse pharmacological effects.However,its effects on melanoma remain unclear.Cellular prolif-eration assays,multi-omics analyses,and xenograft models were used to determine the effect of TET on melanoma.The direct target of TET was identified using biotin-TET pull-down liquid chromatograph-mass spectrometry(LC-MS),cellular thermal shift assays,and isothermal titration calorimetry(ITC)analysis.Our findings revealed that TET treatment induced robust cellular autophagy depending on activating transcription factor 6(ATF6)-mediated endoplasmic reticulum(ER)stress.Simultaneously,it hindered autophagic flux by inducing cytoskeletal protein depolymerization in melanoma cells.TET treatment resulted in excessive accumulation of reactive oxygen species(ROS)and simultaneously triggered mitophagy.Sirtuin 5(SIRT5)was ultimately found to be a direct target of TET.Mechanistically,TET led to the degradation of SIRT5 via the ubiquitin(Ub)-26S proteasome system.SIRT5 knockdown induced ROS accumulation,whereas SIRT5 overexpression attenuated the TET-induced ROS accumula-tion and autophagy.Importantly,TET exhibited anti-cancer effects in xenograft models depending on SIRT5 expression.This study highlights the potential of TET as an antimelanoma agent that targets SIRT5.These findings provide a promising avenue for the use of TET in melanoma treatment and underscore its potential as a therapeutic candidate.

Objective To investigate the effects and molecular mechanism of Notoginsenoside R1 on hypobaric hypoxia-induced high-altitude pulmonary edema based on nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/glutathione peroxidase 4(GPX4)signaling pathway.Methods Forty-two 6-week-old male SD rats were acclimatized and fed for one week,and then randomized into the Control group,negative control group(NGR1,100 mg·kg-1),model group(HH),Notoginsenoside R1 low dose group(NGR1-low,50 mg·kg-1),Notoginsenoside R1 high dose group(NGR1-high,100 mg·kg-1),and dexamethasone group(Dex,4 mg·kg-1),with seven rats in each group.After the drug administration group was administered with Notoginsenoside R1 or dexamethasone by intraperitoneal injection,the hypobaric hypoxia simulation chamber was used to simulate the plateau environment for modeling.Detect the total protein concentration in bronchoalveolar lavage fluid(BALF)and measure the lung wet/dry weight ratio(W/D);ELISA to measure inflammation levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β in BALF;hematoxylin-eosin(HE)staining to visualize pathomorphologic changes in the lungs of each group;biochemical kit detect the content and activity of reactive oxygen species(ROS),superoxide dismutase(SOD),and reduced glutathione(GSH)in the lung tissues;Western blot to detect the protein expression of Nrf2,HO-1,GPX4,SLC7A11 and NOX1 in lung tissues.Results Compared with the Control group,the total protein concentration in the BALF of rats in the HH group was markedly increased(P<0.01),and the lung W/D was markedly increased(P<0.01);lung histopathology showed obvious thickening of the alveolar wall,interstitial edema,hemorrhage and massive inflammatory exudation in the alveolar lumen,alveolar septa,and interstitium of the lungs;the levels of TNF-α(P<0.001),IL-6(P<0.01),and IL-1β(P<0.0001)in the BALF were significantly elevated;ROS content was significantly increased(P<0.01),and SOD(P<0.01)and GSH(P<0.01)activities were significantly decreased in lung tissues;the expression of Nrf2 in the nucleus of lung was markedly decreased(P<0.001),and that in the cytoplasm of lung was markedly increased(P<0.05),and the expression of HO-1(P<0.01),GPX4(P<0.01),and SLC7A11(P<0.05)was significantly decreased,and that of NOX1 was significantly increased(P<0.01).Compared with the HH group,the total protein concentration in the BALF of rats in each drug intervention group was significantly reduced,and the lung W/D was significantly reduced;the lung histopathology was significantly improved;the levels of TNF-α,IL-6,and IL-1β in the BALF were significantly decreased;the content of ROS in the lung tissues was significantly reduced,and the activities of SOD and GSH were significantly elevated;the expression of Nrf2 in the nucleus of lung was markedly increased,the expression of Nrf2 in the cytoplasm of lung was markedly decreased,the expression of HO-1,GPX4,SLC7A11 was markedly increased,and the expression of NOX1 was markedly decreased.Conclusion Notoginsenoside R1 inhibits hypobaric hypoxia-induced ferroptosis and attenuates pulmonary edema,lung inflammation and oxidative stress by a mechanism that may be related to the regulation of the Nrf2/HO-1/GPX4 pathway.

Objective To investigate the effects and molecular mechanism of Notoginsenoside R1 on hypobaric hypoxia-induced high-altitude pulmonary edema based on nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/glutathione peroxidase 4(GPX4)signaling pathway.Methods Forty-two 6-week-old male SD rats were acclimatized and fed for one week,and then randomized into the Control group,negative control group(NGR1,100 mg·kg-1),model group(HH),Notoginsenoside R1 low dose group(NGR1-low,50 mg·kg-1),Notoginsenoside R1 high dose group(NGR1-high,100 mg·kg-1),and dexamethasone group(Dex,4 mg·kg-1),with seven rats in each group.After the drug administration group was administered with Notoginsenoside R1 or dexamethasone by intraperitoneal injection,the hypobaric hypoxia simulation chamber was used to simulate the plateau environment for modeling.Detect the total protein concentration in bronchoalveolar lavage fluid(BALF)and measure the lung wet/dry weight ratio(W/D);ELISA to measure inflammation levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β in BALF;hematoxylin-eosin(HE)staining to visualize pathomorphologic changes in the lungs of each group;biochemical kit detect the content and activity of reactive oxygen species(ROS),superoxide dismutase(SOD),and reduced glutathione(GSH)in the lung tissues;Western blot to detect the protein expression of Nrf2,HO-1,GPX4,SLC7A11 and NOX1 in lung tissues.Results Compared with the Control group,the total protein concentration in the BALF of rats in the HH group was markedly increased(P<0.01),and the lung W/D was markedly increased(P<0.01);lung histopathology showed obvious thickening of the alveolar wall,interstitial edema,hemorrhage and massive inflammatory exudation in the alveolar lumen,alveolar septa,and interstitium of the lungs;the levels of TNF-α(P<0.001),IL-6(P<0.01),and IL-1β(P<0.0001)in the BALF were significantly elevated;ROS content was significantly increased(P<0.01),and SOD(P<0.01)and GSH(P<0.01)activities were significantly decreased in lung tissues;the expression of Nrf2 in the nucleus of lung was markedly decreased(P<0.001),and that in the cytoplasm of lung was markedly increased(P<0.05),and the expression of HO-1(P<0.01),GPX4(P<0.01),and SLC7A11(P<0.05)was significantly decreased,and that of NOX1 was significantly increased(P<0.01).Compared with the HH group,the total protein concentration in the BALF of rats in each drug intervention group was significantly reduced,and the lung W/D was significantly reduced;the lung histopathology was significantly improved;the levels of TNF-α,IL-6,and IL-1β in the BALF were significantly decreased;the content of ROS in the lung tissues was significantly reduced,and the activities of SOD and GSH were significantly elevated;the expression of Nrf2 in the nucleus of lung was markedly increased,the expression of Nrf2 in the cytoplasm of lung was markedly decreased,the expression of HO-1,GPX4,SLC7A11 was markedly increased,and the expression of NOX1 was markedly decreased.Conclusion Notoginsenoside R1 inhibits hypobaric hypoxia-induced ferroptosis and attenuates pulmonary edema,lung inflammation and oxidative stress by a mechanism that may be related to the regulation of the Nrf2/HO-1/GPX4 pathway.

Objective:To explore the effect of complications simulated experience in school-age children with type 1 diabetes mellitus.Methods:From March to December 2021, 80 school-age children with type 1 diabetes mellitus who were followed up in the Endocrinology Department of Quanzhou Children's Hospital were selected by convenient sampling. Children were divided into a control group and an observation group using random number method, with 40 cases in each group. The control group received routine treatment and traditional health education, while the observation group implemented complications simulated experience intervention on the basis of the control group. Diabetes Self-Management Questionnaire and Diabetes Knowledge Test were used to evaluate the self-management ability and diabetes knowledge of the two groups of children before and after intervention.Results:The observation group included 37 children, while the control group included 38 children. After intervention, the scores of Diabetes Self-Management Questionnaire and Diabetes Knowledge Test of children in the observation group were higher than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:Complications simulated experience can improve the self-management ability and diabetes knowledge of school-age children with type 1 diabetes mellitus, which is worthy of clinical promotion and practice.