Results of a Pilot External Quality Assessment Scheme for Genetic Testing of Newborns with Spinal Muscular Atrophy.

BACKGROUND: This study aims to evaluate the ability of laboratories to perform spinal muscular atrophy (SMA) genetic testing in newborns based on dried blood spot (DBS) samples, and to provide reference data and advance preparation for establishing the pilot external quality assessment (EQA) scheme for SMA genetic testing of newborns in China. METHODS: The pilot EQA scheme contents and evaluation principles of this project were designed by National Center for Clinical Laboratories (NCCL), National Health Commission. Two surveys were carried out in 2022, and 5 batches of blood spots were submitted to the participating laboratory each time. All participating laboratories conducted testing upon receiving samples, and test results were submitted to NCCL within the specified date. RESULTS: The return rates were 75.0% (21/28) and 95.2% (20/21) in the first and second surveys, respectively. The total return rate of the two examinations was 83.7% (41/49). Nineteen laboratories (19/21, 90.5%) had a full score passing on the first survey, while in the second survey twenty laboratories (20/20, 100%) scored full. CONCLUSIONS: This pilot EQA survey provides a preliminary understanding of the capability of SMA genetic testing for newborns across laboratories in China. A few laboratories had technical or operational problems in testing. It is, therefore, of importance to strengthen laboratory management and to improve testing capacity for the establishment of a national EQA scheme for newborn SMA genetic testing.

A machine learning model for the prediction of down syndrome in second trimester antenatal screening.

BACKGROUND: Down syndrome (DS) is the most common human chromosomal abnormality. About 1200 laboratories carry out antenatal screening for DS in second trimester pregnancies in China. Their prenatal assessment of DS pregnancy risk is based on biometric calculations conducted on maternal serum biochemical markers and ultrasonic markers of fetal growth. However, the performance of this triple test for DS in second trimester pregnancies has a false positive rate of 5%, and a detection rate of about 60%∼65%. METHOD: A total of 58,972 pregnant women, including 49 DS cases, who had undergone DS screening in the second trimester were retrospectively included and a machine learning (ML) model based on random forest was built to predict DS. In addition, the model was applied to another hospital data set of 27,170 pregnant women, including 27 DS cases, to verify the predictive efficiency of the model. RESULTS: The ML model gave a DS detection rate of 66.7%, with a 5% false positive rate in the model data set. In the external verification data set, the ML model achieved a DS detection rate of 85.2%, with a 5% false positive rate . In comparison with the current laboratory risk model, the ML model improves the DS detection rate with the same false positive rate, while the difference has no significance. CONCLUSIONS: The ML model for DS detection described here has a comparable detection rate with the same false positive rate as the DS risk screening software currently used in China. Our ML model exhibited robust performance and good extrapolation, and could function as an alternative tool for DS risk assessment in second trimester maternal serum.

Correlation of serum vitamin A and vitamin E levels with the occurrence and severity of preeclampsia.

OBJECTIVE: To investigate the correlation of serum vitamin A and vitamin E levels with the occurrence and severity of preeclampsia. METHODS: The clinical data of 5,000 pregnant women in our hospital were retrospectively studied. Five hundred and sixty-five pregnant women with preeclampsia were divided into a mild group (259 cases) and a severe group (306 cases). The rest 4435 healthy pregnant women were classified as the healthy group. The concentrations of serum vitamin A and vitamin E in each group were compared to analyze the risk factors for preeclampsia. RESULTS: The levels of vitamin A and vitamin E were negatively correlated with the severity of preeclampsia (P<0.001). The logistic regression analysis showed that older age (OR=1.674), vitamin A deficiency (OR=2.463) and vitamin E deficiency (OR=2.206) were independent risk factors for preeclampsia (all P<0.05). CONCLUSION: Older age, vitamin A deficiency and vitamin E deficiency are the risk factors for preeclampsia. The concentrations of vitamin A and vitamin E are negatively correlated with the severity of preeclampsia. Vitamin A and vitamin E deficiencies in preeclampsia can increase the risk of adverse pregnancy outcomes, which needs timely intervention.

[The value of a novel prenatal diagnosis model with combination of karyotyping and BACs-on-BeadsTM technique].

OBJECTIVE: To explore the value of a novel prenatal diagnosis model using combined chromosomal karyotyping and BACs-on-BeadsTM(BoBs), a newly-developed technique. METHODS: 1048 single pregnancy pregnant women with various diagnostic indications were performed amniocentesis for prenatal diagnosis with karyotyping and BoBs simultaneously. RESULTS: Among 1047 successfully cultured specimens, 50 chromosomal abnormalities were identified with BoBs, including 43 common chromosomal trisomies, 3 chimeric chromosomes and 4 structural abnormalities, of which 3 microdeletions/microduplications were not detected with karyotyping. Except for extra yield of 1 Robertsonian translocation, the other numerical chromosomal abnormalities were detected with both karyotyping and BoBs. Ten fetal chromosome abnormalities were confirmed with karyotyping, including 8 structural abnormalities and 2 chimeric chromosomes. CONCLUSION: Combination of karyotyping and BoBs turned out to be a rapid, complementary and effective diagnostic model for fetal chromosomal abnormalities and microdeletion syndromes, which could yield a higher detection rate of fetal chromosomal abnormalities and chromosomal microdeletions/microduplications.

Nitidine chloride inhibits proliferation, induces apoptosis via the Akt pathway and exhibits a synergistic effect with doxorubicin in ovarian cancer cells.

Nitidine chloride (NC) exhibits anti-tumor properties in various types of tumor. However, to the best of our knowledge there is no previous evidence of NC involvement in the apoptosis or proliferation of ovarian cancer cells and the underlying molecular mechanisms. The present study aimed to investigate the influence of NC on the viability and apoptosis of ovarian cancer cells and the synergistic effect NC and doxorubicin (DOX) may have on ovarian cancer cells. The viability and proliferation of ovarian cancer cells were examined using a methyl thiazolyl tetrazolium assay and 3H-thymidine incorporation assay. The apoptotic rate of ovarian cancer cells was detected by flow cytometry. The expression of apoptosis­associated proteins and Akt serine/threonine kinase 1 (Akt) were determined by western blot analysis following NC treatment. The inhibitory effect of NC on the proliferation of ovarian cancer cells was demonstrated in a time and dose­dependent manner. The pro-apoptotic effect of NC on ovarian cancer cells was also observed. It was determined that NC significantly downregulated the protein expression levels of B­cell CLL/lymphoma 2 (Bcl-2) and upregulated the expression of Bcl­2­associated X protein, p53, caspase­3 and ­9. NC suppressed Akt phosphorylation. Additionally, the present study demonstrated that the effect of NC on the proliferation and apoptosis of ovarian cancer cells was Akt­dependent by using the phosphatidylinositol-4,5-bisphosphate 3-kinase/Akt signaling pathway inhibitor, LY294002. NC exhibited a synergistic inhibitory effect on the viability of ovarian cancer cells when combined with DOX. The current study demonstrated that NC inhibited the proliferation and induced the apoptosis of ovarian cancer cells via the Akt signaling pathway and highlighted its potential clinical application for the treatment of ovarian cancer.

Nitidine chloride inhibits ovarian cancer cell migration and invasion by suppressing MMP-2/9 production via the ERK signaling pathway.

Nitidine chloride (NC) has been demonstrated to exert anti-tumor effects on various types of tumor. However, no studies have investigated the anti­metastatic effect of NC on ovarian cancer cells, and the underlying mechanisms have not yet been clearly established. The present study aimed to determine the effect of NC on the migration and invasion of ovarian cancer cells. Cell viability and proliferation of ovarian cancer cells were assessed by MTT assay. A scratch wound healing assay and Transwell assays were performed to detect migration and invasion of cells, respectively. The expression levels of matrix metalloproteinase (MMP)­2 and 9 were detected at the mRNA and protein level following stimulation with NC. Subsequently, the expression of mitogen­activated protein kinases was detected by western blot analysis. Finally, an inhibitor of extracellular signal­regulated kinase (ERK) was applied to investigate the effect of NC on the expression of MMP­2/9 as well as the migration and invasion of cells. It was found that NC suppressed the proliferation, migration and invasion of A2780 ovarian cancer cells. NC downregulated MMP­2 and MMP­9 in a dose­ and time­dependent manner. In addition, NC was also able to downregulate phosphorylation of ERK. Furthermore, by applying an ERK inhibitor, U0126, the effect of NC on the expression of MMP-2/9 and inhibition of cell migration and invasion was verified. Taken together, these results demonstrated that NC inhibited the migration and invasion of ovarian cancer cells via the ERK signaling pathway.