Objective To observe the effect of Huoxin pill （concentrated pill） combined with Baduanjin on the prognosis of patients with acute myocardial infarction complicated by percutaneous coronary intervention （PCI）. Methods 120 patients with acute myocardial infarction complicated with heart failure who received emergency interventional treatment in our hospital from January 2022 to January 2023 were randomly divided into western medicine treatment group and Traditional Chinese Medicine （TCM）comprehensive treatment group. Western medicine treatment: standard western medicine treatment + five prescriptions for cardiac rehabilitation; based on western medicine treatment, Huoxin pill （concentrated pill） combined with Baduanjin therapy was added to the TCM comprehensive treatment group, and the follow-up was 6 months. The observed indexes were exertion angina pectoris scale, Chinese Medicine Syndrome Scale, Chinese medicine syndrome treatment effect evaluation, 6-minute walking test （6MWT）, left ventricular ejection fraction （LVEF）, and brain natriuretic peptide precursor （pro-BNP）. Results Sixty patients were enrolled in the two groups respectively. During the treatment, 2 cases fell off in the western medicine treatment group and 8 cases fell off in the TCM comprehensive treatment group, and a total of 110 cases were enrolled in the group. Compared with the western treatment group, TCM combined therapy significantly improved angina pectoris scale score, TCM Syndrome Scale score, pro-BNP, LVEF and 6MWT （P<0.001）. There were no significant differences in blood routine, liver and kidney function, potassium, blood glucose, blood lipids and cardiac Troponin I （cTnI） between the two groups （P>0.05）. No adverse cardiovascular events occurred during the entire treatment period in both groups. Conclusion Huoxin pill （concentrated pill） combined with Baduanjin was more effective than western therapy in improving LVEF, 6MWT distance, exercise tolerance and cardiac function in patients with acute myocardial infarction complicated with heart failure.

Objective: To investigate the biocompatibility of copper-gadolinium nanomaterials and their antibacterial efficacy againstEscherichia coli(E.coli). Methods: Copper-gadolinium nanomaterials were prepared using a hydrothermal method, and the samples were characterized and analyzed using instruments such as transmission electron microscopy. The biocompatibility of copper-gadolinium nanomaterials was evaluated through CCK-8 experiments with mouse fibroblast cells(L929). Copper-gadolinium nanomaterials were co-cultured withE.coli, and their antibacterial performance was observed and assessed. Results: Transmission electron microscopy and Fourier transform infrared spectroscopy analysis confirmed the synthesis of the copper-gadolinium nanomaterials. The CCK-8 experiment results showed that the material had reliable biocompatibility within a concentration range of 0-2 μg/ml. The antibacterial experiments verified the excellent anti-E.coliactivity of copper-gadolinium, with the antibacterial effect positively correlating with concentration and co-cultivation time. When copper-gadolinium nanomaterials at a concentration of 2 μg/ml were co-cultured withE.colifor 4 hours, the antibacterial rate reached 99.99%. Conclusion The 2 μg/ml copper-gadolinium nanomaterials exhibit reliable biocompatibility and excellent anti-E.colicapabilities.

OBJECTIVES: To investigate the genotypes and biochemical phenotypes of neonates with abnormal metabolism of butyrylcarnitine (C4). METHODS: One hundred and twenty neonates with increased C4 levels detected by tandem mass spectrometry in the neonatal screening at Children's Hospital, Zhejiang University School of Medicine from January 2018 to June 2023 were included. The initial screening data and recalled data of C4 and C4/C3 were collected and converted into multiples of C4 reference range. Next generation sequencing was performed and the exons with adjacent 50 bp regions of ACAD8 and ACADS genes were captured by liquid phase capture technique. Variant information was obtained by bioinformatic analysis and the pathogenicity were classified according to the American College of Medical Genetics and Genomics criteria. The Wilcoxon rank sum test was used to analyze the differences in C4 levels among neonates with different variation types. RESULTS: In total, 32 variants in ACAD8 gene were detected, of which 7 variants were reported for the first time; while 41 variants of ACADS gene were detected, of which 17 variants have not been previously reported. There were 39 cases with ACAD8 biallelic variations and 3 cases with ACAD8 monoallelic variations; 34 cases with ACADS biallelic variations and 36 cases with ACADS monoallelic variations. Furthermore, 5 cases were detected with both ACAD8 and ACADS gene variations. Inter group comparison showed that the multiples of C4 reference range in initial screening and re-examination of the ACAD8 biallelic variations and ACADS biallelic variations groups were significantly higher than those of the ACADS monoallelic variations group (all P<0.01), while the multiples in the ACAD8 biallelic variations group were significantly higher than those in the ACADS biallelic variations group (all P<0.01). The multiples of C4 reference range in the initial screening greater than 1.5 times were observed in all neonates carrying ACAD8 or ACADS biallelic variations, while only 25% (9/36) in neonates carrying ACADS monoallelic variations. CONCLUSIONS ACAD8 and/or ACADS gene variants are the main genetic causes for elevated C4 in newborns in Zhejiang region with high genotypic heterogeneity. The C4 levels of neonates with biallelic variations are significantly higher than those of neonates with monoallelic variations. The cut-off value for C4 level could be modestly elevated, which could reduce the false positive rate in tandem mass spectrometry neonatal screening.
Child ; Humans ; Infant, Newborn ; Acyl-CoA Dehydrogenase/genetics* ; Genotype ; Phenotype ; Carnitine/metabolism* ; Mutation

Newborn screening (NBS) plays a significant role in reducing the risk of birth defects. NBS in China began in the early 1980s. Under the protection of laws and regulations and the leadership of the national health administration, approved screening centers in public hospitals took the responsibility for publicity, screening, diagnosis, treatment, follow-up and management of birth defects. As of 2022, 31 provinces (autonomous regions and municipalities directly under the central government) have carried out NBS for phenylketonuria, congenital hypothyroidism, and hearing loss, 23 provinces have carried out screening for glucose-6-phosphate dehydrogenase (with a screening rate of 89.24%), and 24 provinces have carried out screening for congenital adrenal cortical hyperplasia (91.45% screening rate). Over the past four decades, screening techniques have evolved from bacterial inhibition, fluorescence analysis, and tandem mass spectrometry for the detection of biochemical markers to genetic testing, which has greatly contributed to the expansion of the types of diseases screened for. The combined use of metabolomics and genomics is currently being explored. Effective management and rigorous quality control of NBS are prerequisites for improving the quality and ensuring the accuracy of screening. The Quality Management System for Newborn Screening System Network (QMS-NBS), established by the National Center for Clinical Laboratories, covers all screening centers and related blood collection agencies. The operation of the QMS-NBS allows the quality and performance of screening to be transparent and measurable, ensuring the quality and efficiency of screening. This article provides an overview of the history of NBS, especially the evolution of policies for the NBS in China, the construction of screening institutions, the number of newborns screened, the incidence rates of screened diseases, the changes in screening technology, the expansion of new diseases screened for, and the quality control of NBS. Overall, the progress in NBS in China has not only benefited from the development and standardization at the technological level, but also benefited from the construction of policies, regulations and ethics.
Infant, Newborn ; Humans ; Neonatal Screening ; Phenylketonurias ; Genetic Testing ; Congenital Hypothyroidism ; China

Objective:To investigate the efficiency of biochemical screening and hotspot gene screening in the detection of neonatal inherited metabolic diseases.Methods:This was a prospective multi-center study.The study was carried out on 21 442 neonatal samples collected from 12 hospitals in 10 provinces from November 2020 to November 2021.The results of biochemical screening and hotspot gene screening were analyzed jointly.Biochemical screening methods included glucose-6-phosphate dehydrogenase deficiency enzyme activity assay and neonatal tandem mass spectrometry.Genetic screening analysis involved 135 genes associated with 75 neonatal diseases.Results:Of all the 21 442 neonates enrolled in the study, 21 205 were subject to biochemical screening.A total of 813 cases were positive in the initial screening, and 0.45% of them (95 cases) were diagnosed after recall.All the 21 442 neonates underwent gene screening.About 168 positive cases were detected in the initial screening, and 0.73% (156 cases) of them were confirmed finally.Biochemical and genetic screening improved the detection sensitivity of such diseases as primary carnitine deficiency, neonatal intrahepatic cholestasis caused by citrin deficiency, and 2-methylbutyrylglycinemia.Moreover, biochemical and genetic screening enabled the detection of more diseases, including the common single-gene genetic diseases such as thalassemia and Wilson disease.Conclusions:In neonatal screening, the combination of biochemical screening and gene screening expands the number of diseases detected and improve screening efficiency.

Objective:To explore the genetic causes of abnormal isovaleryl carnitine (C5) metabolism in newborns.Methods:Retrospective study.The screening and clinical follow-up data of 34 neonates with elevated C5 levels shown by the tandem mass spectrometry test in Children′s Hospital, Zhejiang University School of Medicine from January 2018 to December 2021 were collected.Afterwards, their ethylenediaminetetraacetic acid (EDTA) anticoagulant venous blood was collected to extract genomic DNA.A total of 79 genes related to genetic metabolic diseases, such as ACADSB, IVD and ACADM, were captured by liquid-phase capture technology.High-throughput sequencing and bioinformatics analysis were used to acquire gene variation information and the genes were categorized by American College of Medical Genetics and Genomics classification standard.According to the results of genetic analysis, the newborns with C5 elevation were divided into 3 groups: non-mutation group(11 cases), ACADSB mutation group(16 cases) and IVD mutation group(7 cases). Wilcoxon rank sum test was performed to analyze the difference between these groups. Results:Among 34 neonates, 6 ACADSB variants were detected in 16 cases, and 2 of them [c.461G>A (p.G154E), c.746delC(p.P249Lfs*15)] were novel variants.Eleven IVD variants were detected in 7 cases, and 7 of them [c.118A>G(p.N40D), c.296-10C>G, c.302A>G(p.Y101C), c.537G>A(p.M179I), c.667C>T(p.R223W), c.983A>G(p.K328R), c.1147+ 5G>A] were never reported before.There was no significant difference in the C5 concentration in initial screening among the three groups ( P>0.05). Conclusions:Mutations in ACADSB and IVD genes are the main causes of augmented C5 levels in neonatal screening.For newly discovered genetic variants, functional prediction by multiple bioinformatics analysis software is recommended.And it is also important to carry out clinical follow-up and evaluation.

Objective: To optimize the pretreatment method of N-nitrosamine compounds in ready-to-eat aquatic products. Methods: Market-sold ready-to-eat aquatic products were collected, homogenized and distilled by steam. The samples were extracted for 10 minutes using dispersive liquid-liquid microextraction (DLLME) with ethanol, trichloromethane and sodium chloride (3.0 g). After centrifugation, the organic phase in the lower layer was collected and subjected to gas chromatography-tandem mass spectrometry (GC-MS/MS). The six common N-nitrosamine compounds were determined in ready-to-eat aquatic products using multiple reaction monitoring mode (MRM) and quantified by the internal standard method. Results: The optimized method exhibited a good linear relationship at concentrations of 10.0 to 500 μg/L for determination of 6 N-nitrosamine compounds (correlation coefficient of greater than 0.999), with 0.05 to 0.60 μg/kg limit of detection, 0.15 to 1.60 μg/kg limit of quantitation, mean spiked recovery rates of 71.8% to 108.9%, and relative standard deviations of 1.4% to 8.6%. N-Nitrosodimethylamine showed the highest detection rate in 20 market-sold ready-to-eat aquatic products (90%), and the detection rates of N-Nitrosopyrrolidine, N-Nitrosodiethylamine and N-dibutylnitrosamine were 15%, 10% and 10%, respectively. Conclusion Steam distillation combined with DLLME may optimize the pretreatment method of N-nitrosamine compounds in ready-to-eat aquatic products and meet the measurement requirements.

l-Heptopyranoses are important components of bacterial polysaccharides and biological active secondary metabolites like septacidin (SEP), which represents a group of nucleoside antibiotics with antitumor, antifungal, and pain-relief activities. However, little is known about the formation mechanisms of those l-heptose moieties. In this study, we deciphered the biosynthetic pathway of the l,l-gluco-heptosamine moiety in SEPs by functional characterizing four genes and proposed that SepI initiates the process by oxidizing the 4'-hydroxyl of l-glycero-α-d-manno-heptose moiety of SEP-328 ( 2) to a keto group. Subsequently, SepJ (C5 epimerase) and SepA (C3 epimerase) shape the 4'-keto-l-heptopyranose moiety by sequential epimerization reactions. At the last step, an aminotransferase SepG installs the 4'-amino group of the l,l-gluco-heptosamine moiety to generate SEP-327 ( 3). An interesting phenomenon is that the SEP intermediates with 4'-keto-l-heptopyranose moieties exist as special bicyclic sugars with hemiacetal-hemiketal structures. Notably, l-pyranose is usually converted from d-pyranose by bifunctional C3/C5 epimerase. SepA is an unprecedented monofunctional l-pyranose C3 epimerase. Further in silico and experimental studies revealed that it represents an overlooked metal dependent-sugar epimerase family bearing vicinal oxygen chelate (VOC) architecture.

Objective:To analyze and evaluate the application of quantitative fecal immunochemical test(FIT) in opportunistic screening of colorectal cancer in asymptomatic population undergoing health checkups.Methods:From January 1, 2018 to December 31, 2021, at the Health Management Center of the First Affiliated Hospital of Soochow University, 53 319 subjects who underwent routine health checkups and with quantitative FIT opportunistic screening for colorectal cancer were selected. Those with positive quantitative FIT results and received colonoscopy were enrolled in the FIT positive group, and those with negative quantitative FIT results and received colonoscopy were enrolled in the FIT negative group. The participation rate and positive rate of quantitative FIT were analyzed. The results of colonoscopy and pathological findings were taken as the gold standard, including normal, non-polyposis lesions, polyposis (hyperplastic and(or) inflammatory polyps, non-advanced adenoma, advanced adenoma), and colorectal cancer, the detection rates of various lesions of the FIT positive and negative groups, the quantitative FIT measurement value of subjects, and the sensitivity and negative predictive value of quantitative FIT for colorectal cancer and advanced adenoma were analyzed. The receiver operating characteristic curve (ROC) was drawn and the area under the curve (AUC) was calculated, the screening efficacy of quantitative FIT for colorectal cancer and advanced adenoma was evaluated. Chi-square test or Fisher exact probability method and Wilcoxon rank sum test were used for statistical analysis.Results:A total of 51 420 cases had completed quantitative FIT, and the total participation rate was 96.44% (51 420/53 319). Quantitative FIT was positive in 2 483 cases (4.83%). The participation rate of colonoscopy in FIT positive group was 26.22% (651/2 483), of which 540 cases were enrolled in FIT positive group. The colonoscopy participation rate of FIT negative group was 1.18% (576/48 937), of which 523 cases were enrolled in the FIT negative group. The detection rates of colorectal cancer and advanced adenoma in FIT positive group were both higher than those of the FIT negative group(3.9%, 21/540 vs. 0, 0/523; 16.1%, 87/540 vs. 3.3%, 17/523), and the differences were statistically significant(Fisher exact probability method and χ2=49.79; both P<0.001). Populations with quantitative FIT values from high to low were those with colorectal cancers, advanced adenomas, non-polyp lesions, non-advanced adenomas, normal, and hyperplastic and (or) inflammatory polyps (1 052.0 ng/mL(390.5 ng/mL, 3 058.0 ng/mL); 294.5 ng/mL (116.8 ng/mL, 951.8 ng/mL); 131.5 ng/mL (10.5 ng/mL, 327.3 ng/mL); 97.0 ng/mL (11.0 ng/mL, 238.0 ng/mL); 20.0 ng/mL (0.0 ng/mL, 175.3 ng/mL); 14.0 ng/mL (0.0 ng/mL, 171.0 ng/mL)), and the difference was statistically significant( H=120.53, P<0.001). The sensitivities(95% confidence interval (95% CI)) of quantitative FIT in colorectal cancer and advanced adenoma were 100.0%(80.8% to 100.0%) and 83.6%(74.8% to 89.9%), respectively. The negative predictive values (95% CI) were 100.0%(99.1% to 100.0%) and 96.7%(94.7% to 98.0%), respectively. The results of ROC analysis showed that the AUCs(95% CI) of quantitative FIT in colorectal cancer and advanced adenoma were 0.874(0.820 to 0.928) and 0.723(0.675 to 0.770), respectively. Conclusions:In this study, the participation rate of quantitative FIT is high. More patients with advanced adenomas and colorectal cancers are found in the high risk popolation with positive quantitative FIT. Quantitative FIT has a good sensitivity and a negative predictive value for colorectal cancer and advanced adenoma. Therefore, positive quantitative FIT-colonoscopies sequential screening should be advocated in population undergoing health checkups for colorectal cancer screening, and it may be applicable to large-scale population screening in China.

Objective:To investigate the application feasibility of Region 4 Stork (R4S) system, an international collaborative newborn screening data platform, combined with cut-off value analysis in the neonatal screening for very long chain acyl-CoA dehydrogenase deficiency (VLCADD) by tandem mass spectrometry (MS/MS).Methods:The retrospective study was performed in 2, 040 072 neonates screened by MS/MS in Neonatal Screening Center of Zhejiang Province, China from October 2013 to July 2018. Nine hundred and ten cases were determined and identified as suspected positive VLCADD neonates by traditional cut-off method of tandem mass spectrometry. The original data of these 910 screened neonates were further analyzed by R4S system. Based on clinical diagnosis and ACADL gene test results, the screening efficiency between two methods was statistically compared.Results:The data of 910 suspected VLCADD-positive cases interpreted by cut-off method were further analyzed by R4S system, and the positive interpretation was reduced to 238 cases (including 9 confirmed positive cases). A total of 16 different mutations were found in ACADL gene sequencing among the confirmed children. The screening false positive rate (FPR) declined from 0.44‰ (901/2 040 072) to 0.11‰ (229/2 040 072), the rate of positive predictive value (PPV) increased from 0.99% (9/910) to 3.78% (9/238), and the specificity increased from 99.96% (2 039 162/2 040 063) to 99.99% (2 039 834/2 040 063). There was a statistically significant difference between cut-off method alone and cut-off method combined R4S system analysis (χ2=393.5, P<0.05). Conclusions:The R4S system combined with cut-off method applied in VLCADD neonatal screening by MS/MS can effectively improve screening performance, reduce false positive rate, and has certain value in clinical application.