OBJECTIVE: To explore the genotype-phenotype correlation in a Chinese family with structural brain abnormalities due to variant of the TUBB gene. METHODS: A family undergoing prenatal diagnosis at Beijing Obstetrics and Gynecology Hospital in October 2024 was selected as the study subject. Clinical data were collected. Amniotic fluid sample was subjected to chromosomal copy number variation sequencing (CNV-seq). Trio whole-exome sequencing (Trio-WES) was carried out on the amniotic fluid and parental blood samples, and candidate variant was verified by Sanger sequencing. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: 2023-KY-076-01). RESULTS: Both prenatal ultrasound and fetal MRI showed deviation of brain midline, unilateral lateral ventriculomegaly, and bilateral gyral asymmetry. Trio-WES revealed that the fetus has harbored a maternally derived heterozygous missense variant of the TUBB gene [NM_178014.4: c.155A>G (p.N52S)]. Sanger sequencing confirmed that the woman and a previously terminated fetus both harbored the same variant. Both the proband and two fetuses exhibited similar neuroimaging abnormalities including midline deviation and asymmetrical gyri. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as likely pathogenic (PM2_Supporting+PS2_Moderate+PS3). CONCLUSION The heterozygous c.155A>G (p.N52S) variant was the TUBB gene probably underlay the pathogenesis of the structural brain abnormalities in this family. Above findings have expanded the phenotypic spectrum associated with the variant and facilitated the prenatal diagnosis for this family.
Humans ; Female ; Pregnancy ; Prenatal Diagnosis ; Tubulin/genetics* ; Adult ; Brain/diagnostic imaging* ; Male ; Pedigree ; DNA Copy Number Variations/genetics* ; Exome Sequencing ; Genetic Association Studies ; Magnetic Resonance Imaging

Objective: To explore the setting of gray zone of Treponema pallidum (TP) testing by retrospective analysis of blood donors with single reagent reactive anti-TP results, so as to improve blood utilization and supply safety. Methods: Blood samples were collected from 112 blood donors previously deferred due to single reagent reactive TP antibody results between January 2020 and December 2023, and subjected to dual ELISA reagents and TPPA test. The gray zone panel analysis was performed on the two ELISA reagents currently used in our department. The detection rate at each concentration of the gray zone panle was counted, and the corresponding concentrations for C , C , and C and gray zone cut-off were calculated. Results: Among the 50 samples deferred by reagent 1, 19 were confirmed reactive and 31 non-reactive in supplementary testing. Among the 62 samples deferred by reagent 2, 12 were confirmed reactive and 50 non-reactive in supplementary testing. For reagent 1, the detection rate of was 56% for S/CO≥1 and 20% for 0.5≤S/CO<1, retrospectively. For reagent 2, the detection rate was 27% for S/CO≥1 and 12.5% for 0.5≤S/CO<1, retrospectively. The detection rate for S/CO≥1 was higher than those for 0.5≤S/CO<1 for both reagents. All the 112 samples were negative in TPPA test. The C concentration of reagent 1 was 1.51 mIU/mL, and the concentration range of C ±20% was 1.21-1.81 mIU/mL. The C concentration of reagent 2 was 1.45 mIU/mL, and the concentration range of C ±20% was 1.16-1.74 mIU/mL. The C and C concentration of both reagents were within the C ±20% range, suggesting that the gray zone cutoff for both Reagent 1 and Reagent 2 should be set at S/CO=0.8 (80% of the CO value). Conclusion: All anti-TP single reagent reactive samples with S/CO value within the gray zone was tested negative by TPPA. It is necessary to consider the rationality and necessity of establishing the gray zone, so as to ensure blood safety and improve the utilization rate of blood resources.

Objective:To evaluate the diagnostic performance of 64Cu-prostate specific membrane antigen (PSMA)-Q compared with 18F-FDG in patients with advanced prostate cancer and to analyze the radiation dosimetry of 64Cu-PSMA-Q. Methods:This study was an open-label, single-arm, self-controlled diagnostic evaluation trial. A total of 29 patients (age 58-87 years) with pathologically confirmed advanced prostate cancer in the Affiliated Hospital of North Sichuan Medical College from September 2023 to December 2023 were included. All patients underwent both 64Cu-PSMA-Q PET/CT and 18F-FDG PET/CT examinations. McNemar test was used to compare the detection rates of 64Cu-PSMA-Q PET/CT and 18F-FDG PET/CT for primary lesions, lymph node metastases, and bone metastases. Mann-Whitney U test was applied to compare differences in SUV max and tumor-to-background ratio (TBR) between 64Cu-PSMA-Q PET/CT and 18F-FDG PET/CT. Radiation dosimetry of 64Cu-PSMA-Q PET/CT imaging was performed using OLINDA/EXM 2.1 (adult male model) in 9 patients. Results:Primary lesions were detected in 21 patients. 64Cu-PSMA-Q PET/CT demonstrated a detection rate of 95.2%(20/21) for primary lesions, which was significantly higher than that of 18F-FDG PET/CT (66.7%(14/21); χ2=6.00, P=0.031). Detection rates of lymph node metastases were 65.5%(19/29) for 64Cu-PSMA-Q and 55.2%(16/29) for 18F-FDG, with no significant difference ( χ2=3.00, P=0.250). Similarly, detection rates of bone metastases were 72.4%(21/29) for 64Cu-PSMA-Q and 65.5%(19/29) for 18F-FDG respectively ( χ2=2.00, P=0.500). TBRs on 64Cu-PSMA-Q PET/CT were significantly higher than those on 18F-FDG PET/CT across primary lesions (8.3(2.2, 13.3) vs 2.3(1.0, 5.5); Z=7.16, P=0.002), regional lymph node metastases (4.9(1.4, 8.3) vs 1.7(0.9, 4.0), Z=189.34, P=0.001), and bone metastases (18.7(4.5, 26.9) vs 5.1(2.1, 9.7); Z=24.83, P=0.003). No significant difference in TBR was observed for distant lymph node metastases ( Z=1.49, P=0.135) or benign lesions ( Z=0.91, P=0.558). The whole-body effective dose of 64Cu-PSMA-Q was (28.200±1.590)μSv/MBq among the 9 patients analyzed, with no adverse events related to the tracer observed. Conclusion:64Cu-PSMA-Q is a promising novel PET imaging agent with potential clinical utility for diagnosing prostate cancer and supporting clinical decision-making.

Objective:To evaluate the diagnostic performance of 64Cu-prostate specific membrane antigen (PSMA)-Q compared with 18F-FDG in patients with advanced prostate cancer and to analyze the radiation dosimetry of 64Cu-PSMA-Q. Methods:This study was an open-label, single-arm, self-controlled diagnostic evaluation trial. A total of 29 patients (age 58-87 years) with pathologically confirmed advanced prostate cancer in the Affiliated Hospital of North Sichuan Medical College from September 2023 to December 2023 were included. All patients underwent both 64Cu-PSMA-Q PET/CT and 18F-FDG PET/CT examinations. McNemar test was used to compare the detection rates of 64Cu-PSMA-Q PET/CT and 18F-FDG PET/CT for primary lesions, lymph node metastases, and bone metastases. Mann-Whitney U test was applied to compare differences in SUV max and tumor-to-background ratio (TBR) between 64Cu-PSMA-Q PET/CT and 18F-FDG PET/CT. Radiation dosimetry of 64Cu-PSMA-Q PET/CT imaging was performed using OLINDA/EXM 2.1 (adult male model) in 9 patients. Results:Primary lesions were detected in 21 patients. 64Cu-PSMA-Q PET/CT demonstrated a detection rate of 95.2%(20/21) for primary lesions, which was significantly higher than that of 18F-FDG PET/CT (66.7%(14/21); χ2=6.00, P=0.031). Detection rates of lymph node metastases were 65.5%(19/29) for 64Cu-PSMA-Q and 55.2%(16/29) for 18F-FDG, with no significant difference ( χ2=3.00, P=0.250). Similarly, detection rates of bone metastases were 72.4%(21/29) for 64Cu-PSMA-Q and 65.5%(19/29) for 18F-FDG respectively ( χ2=2.00, P=0.500). TBRs on 64Cu-PSMA-Q PET/CT were significantly higher than those on 18F-FDG PET/CT across primary lesions (8.3(2.2, 13.3) vs 2.3(1.0, 5.5); Z=7.16, P=0.002), regional lymph node metastases (4.9(1.4, 8.3) vs 1.7(0.9, 4.0), Z=189.34, P=0.001), and bone metastases (18.7(4.5, 26.9) vs 5.1(2.1, 9.7); Z=24.83, P=0.003). No significant difference in TBR was observed for distant lymph node metastases ( Z=1.49, P=0.135) or benign lesions ( Z=0.91, P=0.558). The whole-body effective dose of 64Cu-PSMA-Q was (28.200±1.590)μSv/MBq among the 9 patients analyzed, with no adverse events related to the tracer observed. Conclusion:64Cu-PSMA-Q is a promising novel PET imaging agent with potential clinical utility for diagnosing prostate cancer and supporting clinical decision-making.

Objective To explore the biological mechanisms of isorhamnetin(ISO)inhibition of breast cancer(BC)progression and effects on paclitaxel sensitivity by in vivo and in vitro experiments.Methods The effects of ISO on MDA-MB-231 cell viability,migration,invasion and apoptosis were explored by CCK-8,scratch,Transwell invasion and apoptosis assays.ISO biological functions were analyzed and core target genes were identified by genome-wide microarrays,functional enrichment and protein-protein interactions(PPI).CYP1B1 expression characterization and prognosis in BC were assessed by quantitative Real-time polymerase chain reaction(qRT-PCR),Western blotting(WB),and Kaplan-Meier plotter database.The effects of CYP1B1 overexpression on MDA-MB-231 cell viability,migration,invasion and apoptosis were explored by CCK-8,scratch,Transwell invasion and apoptosis assays.The effects of ISO and CYP1B1 on BC tumor growth were analyzed by establishing a subcutaneous graft tumor animal model and verified by histological analysis with immunohistochemistry(IHC)and hematoxylin and eosin(HE)staining.The effects of ISO and CYP1B1 on NF-κB signaling were analyzed by qRT-PCR and WB.The effect of ISO combined with paclitaxel on BC progression was analyzed by in vitro and in vivo experiments.Results The results of CCK-8 assay showed that ISO inhibited the viability of MDA-MB-231 cells with an IC50 value of 11.93 μmol/L;the scratch,Transwell invasion and apoptosis assays confirmed that ISO inhibited the migration and invasion of MDA-MB-231 cells and promoted apoptosis(P＜0.05).Volcano plot showed a total of 80 differentially expressed genes(DEGs),of which ISO promoted the expression of 27 genes and inhibited the expression of 53 genes.Gene set enrichment analyses showed that DEGs were mainly enriched in the signaling of organic hydroxyl compound metabolic process,inflammatory response,sterol metabolic process,and nuclear factor-κB(NF-κB).qRT-PCR,WB,and Kaplan-Meier plotter results showed that CYP1B1 mRNA and protein expression was increased in BC and mediated worse prognosis(P＜0.05).CCK-8,scratch,Transwell invasion and apoptosis assays showed that CYP1B1 overexpression enhanced MDA-MB-231 cell viability,migration and invasion,and inhibited apoptosis,which was reversed by the addition of ISO(P＜0.05).The results of in vivo experiments showed that ISO downregulated CYP1B1 expression and attenuated NF-κB signaling(P＜0.05).The results of in vitro and in vivo experiments showed that ISO combined with paclitaxel significantly inhibited BC cell viability and tumor growth(P＜0.05).Conclusion ISO inhibits BC malignant biological behavior by modulating the CYP1B1/NF-κB signaling axis,and ISO enhances paclitaxel sensitivity.

Objective To explore the biological mechanisms of isorhamnetin(ISO)inhibition of breast cancer(BC)progression and effects on paclitaxel sensitivity by in vivo and in vitro experiments.Methods The effects of ISO on MDA-MB-231 cell viability,migration,invasion and apoptosis were explored by CCK-8,scratch,Transwell invasion and apoptosis assays.ISO biological functions were analyzed and core target genes were identified by genome-wide microarrays,functional enrichment and protein-protein interactions(PPI).CYP1B1 expression characterization and prognosis in BC were assessed by quantitative Real-time polymerase chain reaction(qRT-PCR),Western blotting(WB),and Kaplan-Meier plotter database.The effects of CYP1B1 overexpression on MDA-MB-231 cell viability,migration,invasion and apoptosis were explored by CCK-8,scratch,Transwell invasion and apoptosis assays.The effects of ISO and CYP1B1 on BC tumor growth were analyzed by establishing a subcutaneous graft tumor animal model and verified by histological analysis with immunohistochemistry(IHC)and hematoxylin and eosin(HE)staining.The effects of ISO and CYP1B1 on NF-κB signaling were analyzed by qRT-PCR and WB.The effect of ISO combined with paclitaxel on BC progression was analyzed by in vitro and in vivo experiments.Results The results of CCK-8 assay showed that ISO inhibited the viability of MDA-MB-231 cells with an IC50 value of 11.93 μmol/L;the scratch,Transwell invasion and apoptosis assays confirmed that ISO inhibited the migration and invasion of MDA-MB-231 cells and promoted apoptosis(P＜0.05).Volcano plot showed a total of 80 differentially expressed genes(DEGs),of which ISO promoted the expression of 27 genes and inhibited the expression of 53 genes.Gene set enrichment analyses showed that DEGs were mainly enriched in the signaling of organic hydroxyl compound metabolic process,inflammatory response,sterol metabolic process,and nuclear factor-κB(NF-κB).qRT-PCR,WB,and Kaplan-Meier plotter results showed that CYP1B1 mRNA and protein expression was increased in BC and mediated worse prognosis(P＜0.05).CCK-8,scratch,Transwell invasion and apoptosis assays showed that CYP1B1 overexpression enhanced MDA-MB-231 cell viability,migration and invasion,and inhibited apoptosis,which was reversed by the addition of ISO(P＜0.05).The results of in vivo experiments showed that ISO downregulated CYP1B1 expression and attenuated NF-κB signaling(P＜0.05).The results of in vitro and in vivo experiments showed that ISO combined with paclitaxel significantly inhibited BC cell viability and tumor growth(P＜0.05).Conclusion ISO inhibits BC malignant biological behavior by modulating the CYP1B1/NF-κB signaling axis,and ISO enhances paclitaxel sensitivity.

Purpose To investigate whether ketogenic diet(KD)can promote cognition by regulating brain metabolism and neuroinflammation in Alzheimer's disease model mice.Materials and Methods Twenty male APP/PS1 mice were randomly assigned to either a KD group(APP/PS1+KD)or a regular diet group(APP/PS1),with 10 mice in each group.Additionally,10 wild-type C57BL/6 male mice served as the control group.The APP/PS1+KD group was fed with a ketogenic feed,the APP/PS1 group received a regular diet,and the control group was maintained on standard chow for a duration of 4 months.Blood ketone levels of mice were monitored after 4 weeks and 4 months of continuous feeding.Cognitive function was assessed via the morris water maze.18F-FDG and 18F-DPA-714 micro PET/CT were performed to evaluate the effects of KD on glucose metabolism and neuroinflammation across various brain regions in the Alzheimer's disease mice.Following PET/CT imaging,brain tissue samples were collected,and the hippocampal Cal region was selected for paraffin sectioning to detect the expression of glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1 through immunofluorescence analysis.Results In the Morris water maze in the fourth month,compared with the control group,the APP/PS1 group had a significantly longer escape latency on days 3-4(P＜0.05 or P＜0.01).Compared with the control group,the APP/PS1 group showed a significant decrease in relative 18F-FDG uptake in brain regions such as the striatum,hippocampus,dorsal thalamus,central gray matter,superior colliculus,olfactory bulb,and midbrain(P＜0.01,P＜0.05).Compared with the APP/PS1 group,the APP/PS1+KD group showed a significant increase in relative 18F-FDG uptake in the hippocampus and dorsal thalamus(P＜0.01).Compared with the control group,the APP/PS1 group showed a significant increase in relative uptake of 18F-DPA-714 in brain regions such as the striatum,hippocampus and hypothalamus(P＜0.05 or P＜0.001).Compared with the APP/PS1 group,the APP/PS1+KD group decreased the relative uptake of 18F-DPA-714 in the hippocampus(P＜0.01).Compared with the control group and APP/PS1+KD group,the fluorescence intensity of glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1 protein in the brain(hippocampus)of APP/PS1 group mice was significantly increased(both P＜0.01).Conclusion KD has the potential to ameliorate cognitive and behavioral deficits in APP/PS1 mice by enhancing brain metabolism and attenuating neuroinflammation.

Objective:To evaluate the safety of early enteral nutrition (EEN) support in patients with severe intra-abdominal infection and intestinal fistulas.Methods:This was a retrospective cohort study. We collected relevant clinical data of 204 patients with severe intra-abdominal infection and intestinal fistulas who had been managed in the No. 1 Department of General Surgery, Shanghai Ninth People's Hospital, Shanghai Jiaotong University between 1 January 2017 and 1 January 2020. The patients were allocated to EEN or delayed enteral nutrition (DEN) groups depending on whether enteral nutrition had been instituted within 48 hours of admission to the intensive care unit. The primary outcome was 180-day mortality. Other outcomes included rates of intraperitoneal hemorrhage, septic shock, open abdominal cavity, bloodstream infection, mechanical ventilation, and continuous renal replacement therapy. Risk factors for mortality were analyzed by logistic regression.Results:There were no significant differences in hematological data or other baseline characteristics between the two groups at the time of admission to the intensive care unit (all P>0.05). However, septic shock (31.2% [15/48] vs. 15.4% [24/156], χ 2=4.99, P=0.025), continuous renal replacement therapy (27.1% [13/48] versus 9.0% [14/156], χ 2=8.96, P=0.003), and 180-day mortality (31.2% [15/48] vs. 7.7% [12/156], χ 2=15.75, P<0.001) were significantly more frequent in the EEN than the DEN group (all P<0.05). Multivariate regression analysis showed that older age (OR=1.082, 95%CI:1.027-1.139, P=0.003), worse Acute Physiology and Chronic Health Evaluation (APACHE) II scores (OR=1.189, 95%CI: 1.037-1.363, P=0.013), higher C-reactive protein (OR=1.013, 95%CI:1.004-1.023, P=0.007) and EEN (OR=8.844, 95%CI:1.809- 43.240, P=0.007) were independent risk factors for death in patients with severe intra-abdominal infection and intestinal fistulas. Conclusion:EEN may lead to adverse events and increase mortality in patients with both enterocutaneous fistulas and severe abdominal infection. EEN should be implemented with caution in such patients.

Objective:To assess the effectiveness and feasibility of carrier detection for Spinal muscular atrophy (SMA) by using digital PCR assay.Methods:Peripheral blood samples were collected from 214 pregnant women who were routinely screened for SMA carriers, of which 204 were randomly selected samples and 10 were samples with known copy numbers of SMN1 exons 7 and 8. Samples with known copy numbers of SMN1 exons 7 and 8 were randomly mixed into the experiment to validate the performance of the digital PCR assay. The copy numbers of SMN1 exons 7 and 8 and SMN2 exons 7 and 8 in peripheral blood samples were detected by digital PCR assay. The results of SMN1 exons 7 and 8 were compared with those of the quantitative PCR method to assess the reliability and clinical performance of the digital PCR assay. Results:Among the 204 random samples, digital PCR has detected five samples with simultaneous heterozygous deletion of SMN1 exons 7 and 8, three samples with heterozygous deletion of SMN1 exon 8 only, and 196 samples with no deletion of SMN1 exons 7 and 8. Ten samples with known SMN1 exons 7 and 8 copy numbers were detected with the expected values. The digital PCR test results were fully consistent with that of the quantitative PCR. Conclusion:The results of digital PCR for the detection of copy number variation of SMN1 exons 7 and 8 were consistent with qPCR. Digital PCR assay was able to clearly distinguish the copy number of the target genes, therefore can be used for SMA carrier screening. Moreover, it can also detect copy number of SMN2 exons 7 and 8, which can provide more information for genetic counseling.

Objective:To evaluate the safety of early enteral nutrition (EEN) support in patients with severe intra-abdominal infection and intestinal fistulas.Methods:This was a retrospective cohort study. We collected relevant clinical data of 204 patients with severe intra-abdominal infection and intestinal fistulas who had been managed in the No. 1 Department of General Surgery, Shanghai Ninth People's Hospital, Shanghai Jiaotong University between 1 January 2017 and 1 January 2020. The patients were allocated to EEN or delayed enteral nutrition (DEN) groups depending on whether enteral nutrition had been instituted within 48 hours of admission to the intensive care unit. The primary outcome was 180-day mortality. Other outcomes included rates of intraperitoneal hemorrhage, septic shock, open abdominal cavity, bloodstream infection, mechanical ventilation, and continuous renal replacement therapy. Risk factors for mortality were analyzed by logistic regression.Results:There were no significant differences in hematological data or other baseline characteristics between the two groups at the time of admission to the intensive care unit (all P>0.05). However, septic shock (31.2% [15/48] vs. 15.4% [24/156], χ 2=4.99, P=0.025), continuous renal replacement therapy (27.1% [13/48] versus 9.0% [14/156], χ 2=8.96, P=0.003), and 180-day mortality (31.2% [15/48] vs. 7.7% [12/156], χ 2=15.75, P<0.001) were significantly more frequent in the EEN than the DEN group (all P<0.05). Multivariate regression analysis showed that older age (OR=1.082, 95%CI:1.027-1.139, P=0.003), worse Acute Physiology and Chronic Health Evaluation (APACHE) II scores (OR=1.189, 95%CI: 1.037-1.363, P=0.013), higher C-reactive protein (OR=1.013, 95%CI:1.004-1.023, P=0.007) and EEN (OR=8.844, 95%CI:1.809- 43.240, P=0.007) were independent risk factors for death in patients with severe intra-abdominal infection and intestinal fistulas. Conclusion:EEN may lead to adverse events and increase mortality in patients with both enterocutaneous fistulas and severe abdominal infection. EEN should be implemented with caution in such patients.