Objective:To investigate the current status of preimplantation genetic testing for aneuploidies (PGT-A) in China in 2024 and to provide recommendations for ensuring the consistency of PGT-A results.Methods:This study was a nationwide external quality assessment research. The National Center for Clinical Laboratories of the National Health Commission conducted two surveys in June and December 2024, with participation from 31 laboratories across China. During each survey, quality control samples of varying concentrations were distributed: the first survey distributed 6 samples (Batch Nos. 202401-202406) and the second survey distributed 3 samples (Batch Nos. 202431-202433). Participating laboratories were required to submit information including detection platforms, single-cell amplification methods, library construction methods, sequencing instruments, and test results. Collected data were statistically analyzed using Microsoft Excel to evaluate laboratories' PGT-A testing capabilities and determine whether results met predefined quality control requirements.Results:All 31 laboratories submitted results for both surveys. In the first survey, 90.3% (28/31) met qualification criteria with a 9.7% failure rate (3/31). For individual batches, pass rates were 90.3% (28/31) for batches 202401-202403 and 100% (31/31) for batches 202404-202406. The second survey achieved 100% (31/31) compliance across all three batches (202431-202433), with each batch maintaining a 100% pass rate.Conclusion:The detection quality of PGT-A in China is generally good, but with differences, and factors such as sample concentration, transportation conditions and detection methods affect the consistency of the results. The study highlights the urgent need for nationwide external quality assessment of PGT-A in China. This will help identify issues in laboratories in a timely manner and ensure the consistency of PGT-A test results.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To investigate the dosimetric impact of dwell position spacing in the design of three-dimensional (3D) interstitial brachytherapy plans for cervical cancer, and to provide a reference for selecting dwell spacing in clinical planning.Methods:A total of 15 patients with cervical cancer who underwent 3D interstitial brachytherapy at Tianjin Medical University Cancer Institute & Hospital between March 2022 and March 2024 were selected using simple random sampling. For each patient, 10 brachytherapy plans were generated with different dwell position spacings set at 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 mm, respectively. Key parameters among different dwell spacings compared included D 90%, V 100%, V 200%, and V 300% for the high-risk clinical target volume (HRCTV); D 90% for the intermediate-risk clinical target volume (IRCTV); D 2 cm3 for organs at risk (OARs) (bladder, small intestine, colon, and rectum); and the total dwell time. Statistical analyses were performed using repeated measurement ANOVA or the Friedman test. Results:Among different dwell spacings, there were no statistically significant differences in HRCTV D 90%, HRCTV V 100%, bladder D 2 cm3, and rectum D 2 cm3 among different dwell spacings ( P=0.075, 0.061, 0.480, 0.639). All plans with dwell spacings ≤ 3 mm met clinical dose requirements. When the dwell spacing was set to 1 mm, HRCTV V 200% and V 300% had the smallest mean values, while IRCTV D 90% and total dwell time had the largest mean values; all differences were statistically significant ( P<0.05). When the dwell spacing was ≥6 mm, an increase in spacing led to a decrease in mean small intestine D 2 cm3, and total dwell time, but an increase in HRCTV V 200% and a decrease in IRCTV D 90%, with statistically significant differences compared to spacings of 1-4 mm ( P<0.05). When the dwell spacing was ≥8 mm, the median colon D 2 cm3 decreased, with statistically significant differences compared to spacings of 1-3 mm ( P<0.05). Conclusions:For 3D interstitial brachytherapy planning in cervical cancer, dwell position spacings ≤ 3 mm can meet clinical dose requirements, with 1 mm providing optimal target coverage. Spacings ≥6 mm / ≥8 mm can reduce radiation dose to the small intestine and colon, respectively, while also shortening dwell time.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To investigate the dosimetric impact of dwell position spacing in the design of three-dimensional (3D) interstitial brachytherapy plans for cervical cancer, and to provide a reference for selecting dwell spacing in clinical planning.Methods:A total of 15 patients with cervical cancer who underwent 3D interstitial brachytherapy at Tianjin Medical University Cancer Institute & Hospital between March 2022 and March 2024 were selected using simple random sampling. For each patient, 10 brachytherapy plans were generated with different dwell position spacings set at 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 mm, respectively. Key parameters among different dwell spacings compared included D 90%, V 100%, V 200%, and V 300% for the high-risk clinical target volume (HRCTV); D 90% for the intermediate-risk clinical target volume (IRCTV); D 2 cm3 for organs at risk (OARs) (bladder, small intestine, colon, and rectum); and the total dwell time. Statistical analyses were performed using repeated measurement ANOVA or the Friedman test. Results:Among different dwell spacings, there were no statistically significant differences in HRCTV D 90%, HRCTV V 100%, bladder D 2 cm3, and rectum D 2 cm3 among different dwell spacings ( P=0.075, 0.061, 0.480, 0.639). All plans with dwell spacings ≤ 3 mm met clinical dose requirements. When the dwell spacing was set to 1 mm, HRCTV V 200% and V 300% had the smallest mean values, while IRCTV D 90% and total dwell time had the largest mean values; all differences were statistically significant ( P<0.05). When the dwell spacing was ≥6 mm, an increase in spacing led to a decrease in mean small intestine D 2 cm3, and total dwell time, but an increase in HRCTV V 200% and a decrease in IRCTV D 90%, with statistically significant differences compared to spacings of 1-4 mm ( P<0.05). When the dwell spacing was ≥8 mm, the median colon D 2 cm3 decreased, with statistically significant differences compared to spacings of 1-3 mm ( P<0.05). Conclusions:For 3D interstitial brachytherapy planning in cervical cancer, dwell position spacings ≤ 3 mm can meet clinical dose requirements, with 1 mm providing optimal target coverage. Spacings ≥6 mm / ≥8 mm can reduce radiation dose to the small intestine and colon, respectively, while also shortening dwell time.

Objective:To investigate the current status of preimplantation genetic testing for aneuploidies (PGT-A) in China in 2024 and to provide recommendations for ensuring the consistency of PGT-A results.Methods:This study was a nationwide external quality assessment research. The National Center for Clinical Laboratories of the National Health Commission conducted two surveys in June and December 2024, with participation from 31 laboratories across China. During each survey, quality control samples of varying concentrations were distributed: the first survey distributed 6 samples (Batch Nos. 202401-202406) and the second survey distributed 3 samples (Batch Nos. 202431-202433). Participating laboratories were required to submit information including detection platforms, single-cell amplification methods, library construction methods, sequencing instruments, and test results. Collected data were statistically analyzed using Microsoft Excel to evaluate laboratories' PGT-A testing capabilities and determine whether results met predefined quality control requirements.Results:All 31 laboratories submitted results for both surveys. In the first survey, 90.3% (28/31) met qualification criteria with a 9.7% failure rate (3/31). For individual batches, pass rates were 90.3% (28/31) for batches 202401-202403 and 100% (31/31) for batches 202404-202406. The second survey achieved 100% (31/31) compliance across all three batches (202431-202433), with each batch maintaining a 100% pass rate.Conclusion:The detection quality of PGT-A in China is generally good, but with differences, and factors such as sample concentration, transportation conditions and detection methods affect the consistency of the results. The study highlights the urgent need for nationwide external quality assessment of PGT-A in China. This will help identify issues in laboratories in a timely manner and ensure the consistency of PGT-A test results.

Heart failure is one of the main cardiovascular system diseases at present， and it is a clinical syndrome caused by changes in cardiac structure and function， resulting in impaired ejection function or ventricular filling. Therefore， heart failure has become the most important cardiovascular disease in the 21^st century. In recent years， the incidence of heart failure is increasing， and the survival rate of patients with heart failure is very low. Traditional Chinese medicine has rich experience in preventing and treating heart failure. With the modernization of traditional Chinese medicine， more and more attention has been paid to the research， development， and application of active ingredients in traditional Chinese medicine. Traditional Chinese medicine has unique advantages in improving the heart function of patients with heart failure by treating multiple targets and multiple pathways through syndrome differentiation. Astragalus membranacus， a traditional Chinese medicine， is a kind of medicine that benefits Qi and blood circulation and removes evil spirits. It has the functions of improving myocardial energy metabolism and hemodynamics， protecting myocardial muscle， and promoting angiogenesis. Astragalus membranaceus is often used to treat patients with heart failure， yielding remarkable results. In recent years， it has been found that astragaloside， Astragalus polysaccharide， quercetin， calyx isoflavones， and other main active ingredients of Astragalus membranacus can improve cardiac function and treat heart failure by inhibiting inflammatory response， myocardial apoptosis， and myocardial fibrosis. This paper reviewed the research progress of the action and mechanism of the active ingredients of Astragalus membranacus in the treatment of heart failure by studying relevant literature， with a view to providing a reference for its further research， development， and application in the prevention and treatment of heart failure.

ObjectiveTo study the effect and mechanism of Yixintai on mitochondrial fission proteins in the rat model of chronic heart failure. MethodTen of 60 SD rats were randomly selected as the sham operation group， and the remaining 50 rats were subjected to ligation of the left anterior descending coronary artery for the modeling of heart failure post myocardial infarction. The successfully modeled rats were randomized into model， low-， medium-， and high-dose （1.4， 2.8， and 5.6 g·kg^-1， respectively） Yixintai， and trimetazidine （10 mg·kg^-1） groups. The rats were administrated with corresponding doses of drugs by gavage， and the rats in the model group and sham operation group were given an equal volume of normal saline by gavage for 28 consecutive days. Enzyme-linked immunosorbent assay （ELISA） was then employed to measure the levels of amino-terminal pro-B-type natriuretic peptide （NT-pro BNP）， B-type natriuretic peptide （BNP）， and adenosine triphosphate （ATP） in the serum. Color Doppler ultrasound imaging was conducted to examine the cardiac function indicators. Hematoxylin-eosin staining and Masson staining were conducted to observe the pathological changes in the heart， and Image J was used to calculate collagen volume fraction （CVF）. Transmission electron microscopy was employed to observe the ultrastructural changes of myocardial cells. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to measure the apoptosis rate of myocardial cells. Western blot was employed to determine the protein levels of mitochondrial fission protein 1 （Fis1） and mitochondrial fission factor （Mff） in the outer mitochondrial membrane of the myocardial tissue. ResultCompared with the sham operation group， the model group showed elevated levels of NT-pro BNP and BNP in the serum， decreased ATP content， left ventricular ejection fraction （LVEF）， and left ventricular fraction shortening （LVFS）， increased left ventricular end-diastolic diameter （LVIDd） and left ventricular end-systolic diameter （LVIDs）， disarrangement of myocardial cells， inflammatory cell infiltration， increased collagen fibers and CVF， damaged myocardium and mitochondria， and increased apoptosis rate of myocardial cells， and up-regulated expression of Fis1 and Mff in the cardiac tissue （P<0.01）. Compared with the model group， different doses of Yixintai and trimetazidine lowered the serum levels of NT-pro BNP and BNP （P<0.05）， increased the ATP content （P<0.05）， increased LVEF and LVFS （P<0.01）， decreased LVIDd and LVIDs （P<0.01）. Moreover， the drugs alleviated the myocardial inflammatory damage and fibrosis， reduced CVF （P<0.01）， repaired the myocardial mitochondrial structure， and decreased the apoptosis rate of myocardial cells （P<0.01）. Medium- and high-dose Yixintai and trimetazidine down-regulated the expression of Fis1 and Mff in the myocardial tissue （P<0.05）. ConclusionYixintai can improve mitochondrial structure， reduce myocardial cell apoptosis， and improve cardiac function by inhibiting the expression of Fis1 and Mff in the myocardial tissue.

Heart failure is a group of complex clinical syndromes in the middle and late stages of cardiovascular diseases.Mitochondrial homeostasis imbalance is one of the pathological mechanisms in the occurrence and development of heart failure.This article revolved around the"yin-yang theory"in TCM and explained the pathological mechanism of heart failure through mitochondrial homeostasis.Heart failure is the syndrome of deficiency in nature and excess in superficiality fundamental.Its basic pathogenesis is"yang deficiency and yin excess".Based on the deficiency of heart yang qi and the stagnation of yin pathogens,the combination of deficiency and excess runs through the entire disease.Mitochondrial homeostasis imbalance is a manifestation of yin-yang imbalance at the cellular micro level,mainly manifested as inhibition of mitochondrial biosynthesis,mitochondrial dynamics imbalance,mitophagy disorder,etc.,which affects mitochondrial structure and function and leads to abnormal myocardial energy metabolism.Therefore,based on the"yin-yang theory",the basic treatment method is to"tonify deficiency and damage excess"to regulate mitochondrial biosynthesis,mitochondrial dynamics,and mitophagy,thereby maintaining mitochondrial homeostasis and improving myocardial energy metabolism,which is of great significance for the prevention and treatment of heart failure.

Benign prostatic hyperplasia(BPH)is one of the major chronic complications of type 2 diabetes mellitus(T2DM),and sex steroid hormones are common risk factors for the occurrence of T2DM and BPH.The profiles of sex steroid hormones are simultaneously quantified by LC-MS/MS in the clinical serum of patients,including simple BPH patients,newly diagnosed T2DM patients,T2DM complicated with BPH patients and matched healthy individuals.The G protein-coupled estrogen receptor(GPER)inhibitor G15,GPER knockdown lentivirus,the YAP1 inhibitor verteporfin,YAP1 knockdown/overexpression lentivirus,targeted metabolomics analysis,and Co-IP assays are used to investigate the molecular mechanisms of the disrupted sex steroid hormones homeostasis in the pathological process of T2DM complicated with BPH.The homeostasis of sex steroid hormone is disrupted in the serum of patients,accompanying with the proliferated prostatic epithelial cells(PECs).The sex steroid hormone metabolic profiles of T2DM patients complicated with BPH have the greatest degrees of separation from those of healthy individuals.Elevated 17β-estradiol(E2)is the key contributor to the disrupted sex steroid hormone homeostasis,and is significantly positively related to the clinical characteristics of T2DM patients complicated with BPH.Activating GPER by E2 via Hippo-YAP1 signaling exacerbates high glucose(HG)-induced PECs prolifer-ation through the formation of the YAP1-TEAD4 heterodimer.Knockdown or inhibition of GPER-mediated Hippo-YAP1 signaling suppresses PECs proliferation in HG and E2 co-treated BPH-1 cells.The anti-proliferative effects of verteporfin,an inhibitor of YAP1,are blocked by YAP1 overexpression in HG and E2 co-treated BPH-1 cells.Inactivating E2/GPER/Hippo/YAP1 signaling may be effective at delaying the progression of T2DM complicated with BPH by inhibiting PECs proliferation.