[Screening for Duchenne muscular dystrophy in newborns in the Ningxia region]. / å®å¤åœ°åŒºæ–°ç”Ÿå„¿æœæ°è‚Œè¥å »ä¸è‰¯ç—‡ç­›æŸ¥.

OBJECTIVES: To evaluate the incidence rate of Duchenne muscular dystrophy (DMD) in the male newborns in the Ningxia region and establish a critical threshold for screening DMD in newborns to distinguish between the normal population and affected individuals. METHODS: A total of 10 000 male newborns were screened using immunofluorescence analysis of creatine kinase isoenzyme concentrations in heel spot dried blood specimens. Newborns with the concentrations higher than the critical threshold were recalled for serum creatine kinase measurements. Genetic testing was performed to confirm diagnosis in cases showing abnormalities. RESULTS: Among the screened 10 000 male newborns, two were confirmed to have DMD through genetic testing, resulting in a preliminary estimated incidence rate of 1/5 000 for male newborns in the Ningxia region. The critical threshold for creatine kinase isoenzyme concentration in newborns in this region was determined to be 468.57 ng/mL. CONCLUSIONS: Screening for DMD in newborns is feasible in the Ningxia region. Early screening, diagnosis, and treatment of DMD can improve the quality of life for affected individuals and help families make informed decisions regarding further pregnancies.

Metastasis patterns and prognosis in young gastric cancer patients: A propensity score­matched SEER database analysis.

BACKGROUND: Whether young patients with metastatic gastric cancer (GC) had distinct metastasis patterns and survival outcomes from older patients remains controversial. The aim of the present study was to explore the metastasis patterns and prognostic factors in young patients and evaluate the survival outcome in comparison to their older counterparts. MATERIALS AND METHODS: We identified patients with metastatic GC in the surveillance, epidemiology, and end results (SEER) database from 2010 to 2015. The patients were divided into two groups based on age at diagnosis: younger (≤40 years old) and older (>40 years old). We employed the chi-squared test to compare the clinicopathological characteristics between the two age groups. Furthermore, we conducted survival analyses using Kaplan-Meier and Cox regression analyses. To balance disparities in baseline characteristics, we employed propensity score matching (PSM). RESULTS: We identified 5,580 metastatic GC patients from the SEER database, with 237 (4.2%) classified as younger and 5343 (95.8%) as older patients. A total of 237 pairs of patients were generated after adjustment by PSM. Patients in the younger group exhibited a higher proportion of bone-only metastases and a lower proportion of liver-only metastases compared with patients in the older group. Multivariate Cox regression analysis demonstrated that youth was an independent protective factor for overall survival (OS) before and after PSM, but not for gastric cancer-specific survival (GCSS). Among the younger group, patients with liver-only metastasis demonstrated the best prognosis, whereas patients with lung-only metastasis exhibited significantly worse survival outcomes compared with liver-only metastases, even comparable to that of bone metastasis. CONCLUSIONS: Compared with the older group, the metastatic GC patients in the younger group exhibited more aggressive tumors but better prognoses. The metastasis pattern and its effect on the prognosis of GC varied by age group.

Survival benefits of perioperative chemoradiotherapy versus chemotherapy for advanced stage gastric cancer based on directed acyclic graphs.

The overall survival benefits of perioperative chemotherapy (PCT) and perioperative chemoradiotherapy (PCRT) for patients with locally advanced gastric cancer (GC) have not been fully explored. The aim of this study was to compare the benefits of PCT and PCRT in GC patients and determine the factors affecting survival rate using directed acyclic graphs (DAGs). The data of 1,442 patients with stage II-IV GC who received PCT or PCRT from 2000 to 2018 were retrieved from the Surveillance, Epidemiology, and End Results (SEER) database. First, the least absolute shrinkage and selection operator (LASSO) was used to identify possible influencing factors for overall survival. Second, the variables that were selected by LASSO were then used in univariate and Cox regression analyses. Third, corrective analyses for confounding factors were selected based on DAGs that show the possible association between advanced GC patients and outcomes and evaluate the prognosis. Patients who received PCRT had longer overall survival than those who received PCT treatment (P = 0.015). The median length of overall survival of the PCRT group was 36.5 (15.0 - 53.0) months longer than that of the PCT group (34.6 (16.0 - 48.0) months). PCRT is more likely to benefit patients who are aged ≤ 65, male, white, and have regional tumors (P<0.05). The multivariate Cox regression model showed that male sex, widowed status, signet ring cell carcinoma, and lung metastases were independent risk factors for a poor prognosis. According to DAG, age, race, and Lauren type may be confounding factors that affect the prognosis of advanced GC. Compared to PCT, PCRT has more survival benefits for patients with locally advanced GC, and ongoing investigations are needed to better determine the optimal treatment. Furthermore, DAGs are a useful tool for contending with confounding and selection biases to ensure the proper implementation of high-quality research.

Effect of multimodal chemotherapy on survival of gastric cancer with liver metastasis - a population based analysis.

Objectives: Limited efforts have been made to evaluate the effect of multimodal chemotherapy on the survival of gastric cancer patients with liver metastases (LMGC). This study aimed to identify prognostic factors in LMGC patients and the superiority of multimodal chemotherapy with respect to overall survival (OS) in these patients. Methods: We conducted a retrospective cohort study of 1298 patients with M1 stage disease between January 2012 and December 2020. The effects of clinicopathological variables and preoperative chemotherapy (PECT), postoperative chemotherapy (POCT), and palliative chemotherapy on survival in patients with liver metastases (LM group) and non-liver metastases (non-LM group) were compared. Results: Of the 1298 patients analysed, 546 (42.06%) were in the LM group and 752 (57.94%) were in the non-LM group. The median (interquartile range) age was 60 (51-66) years. The 1-year, 3-year and 5-year overall survival (OS) rates in the LM group were 29.3%, 13.9%, and 9.2%, respectively, and those in the non-LM group were. 38.2%, 17.4%, and 10.0%, respectively (P < 0.05, > 0.05, and > 0.05, respectively.) The Cox proportional hazards model revealed that palliative chemotherapy was a significant independent prognostic factor in both the LM and non-LM groups. Age ≥55 years, N stage, and Lauren classification were also independent predictors of OS in the LM group (P < 0.05). Palliative chemotherapy and POCT were associated with improved OS compared with PECT in the LM group (26.3% vs. 36.4% vs. 25.0%, P < 0.001). Conclusion: LMGC patients had a worse prognosis than non- LMGC. Number of metastatic sites more than 1, liver and other metastatic sites, no CT treatment and HER2-negative had a poor prognosis. LMGC patient may benefit more from palliative chemotherapy and POCT than from PECT. Further well-designed, prospective studies are needed to validate these findings.

A rapid and quantitative detection method for plasma soluble growth stimulating gene protein 2 based on time resolved fluorescence immunochromatography.

Background: plasma soluble growth stimulating gene protein 2 (sST2) is a new generation biomarker in heart failure (HF), which is an independent predictor of adverse outcomes of heart failure. Thus, the establishment of a rapid and sensitive method for detecting sST2 is urgently needed. Methods: lanthanide element Eu3+ coated fluorescent nanometer microspheres (Eu3+@FMN) can be used as markers to label monoclonal mouse anti-human sST2 antibody ST-01 (ST-01-Eu3+@FMN). When the immune sandwich complex formed between the monoclonal mouse anti-human sST2 antibody ST-02 and ST-01-Eu3+@FMN on the test band with the appearance of target object sST2, we can detect the fluorescence intensity of Eu3+ on the test band and the quality control band using a dry fluorescence analyzer. We calculated the T/C value (T/C = fluorescence intensity of the test band/fluorescence intensity of the quality control band), fitted to the calibration curve, and measured the concentration value of sST2 in the corresponding sample. Results: the best reaction time was 15 min after condition exploration, and the optimal sample volume was 80 µL. The detection sensitivity of the scheme was 2.14 ng mL-1. The calibration curve of the assay was y = 0.0113x + 0.0033, and the linear range was 5-200 ng mL-1. No cross reaction was found when the samples contained BNP, NT-proBNP, and galectin-3, indicating a good specificity. The precision was good with a relative deviation < 15%. The coefficient of variation of detection results of low-concentration samples and high-concentration samples was 4.20% and 3.30% respectively in the same batch of strip tests, so the intra-assay CV was set as <10%; when different batches of strips were used for testing, the coefficient of variation of detection results of low-concentration samples and high-concentration samples was 10.06% and 8.38% respectively, so the inter-assay CV was set as <15%. Stability test results showed that the relative deviation of test results at each time node was <15%, indicating good stability of the assay strips. The correlation coefficient between the ST-01-Eu3+@FMN based time-resolved fluorescence immunochromatography analysis and sST2 ELISA kit was 0.98. To confirm the usage of our proposed TRF-ICA for clinical samples, it was used to determine the concentration of sST2 in samples obtained from 34 patients with heart insufficiency, acute and chronic heart failure. As a result, we successfully detected a minimal concentration of 5.21 ng mL-1 and a maximum concentration of 184.26 ng mL-1 for sST2. Conclusion: this technique provides a rapid, simple and quantitative detection method for sST2 in clinics. It can help clinicians to predict the incidence of adverse events in patients with HF.

Simultaneous Quantification of Anticardiolipin IgG and IgM by Time Resolved Fluoroimmunoassay.

The autoimmune disease antiphospholipid syndrome (APS) is characterized by the presence of anticardiolipin antibodies (aCL), along with anti-ß2-glycoprotein I (ß2GPI) antibodies and lupus anticoagulant (LA). In this study, we developed a time-resolved fluoroimmunoassay (TRFIA) system for simultaneous quantification of aCL IgG and IgM. A 96-well microtiter plate precoated with the complex of cardiolipin from bovine heart and bovine ß2GPI was incubated with the anticardiolipin IgG and IgM standard substance or serum, and the conjugate of Eu3+-labeled anti-human IgG and Sm3+-labeled anti-human IgM was pipetted to the wells to form a tipical double-antibody-sandwich immunoreactions; finally the fluorescent intensity of Eu3+ and Sm3+ was detected to reflect the quantity of anticardiolipin IgG and IgM. This assay showed a good relationship between fluorescence intensities and the concentration of anticardiolipin antibody(aCL) IgG and IgM, with a low-end sensitivity of 0.1 U/ml for IgG and 0.1 U/ml for IgM, respectively. The intra- and inter-assay coefficients of variation (CV) of the calibrators was 3.0% and 4.51% for IgG, and 2.76% and 4.45% for IgM. The average recovery was 100.38% for aCL IgG and 100.45% for aCL IgM. For serum samples, the results of our method showed a good correlation with those obtained with ELISA kit. Simultaneous detection of aCL-IgG and aCL-IgM in the same reaction well can optimize assay performance by avoiding potential influence of different reaction conditions-timing, and well-to-well difference in concentration and characteristics of cardiolipin antigen. The results of a combo aCL-IgG and aCL-IgM assay for the same sample are more consistent and more reliable. This dual-label time-resolved fluoroimmunoassay is sensitive for detecting aCL IgG and IgM across a wide concentration range with stable reagents and may assist in the clinical diagnosis of antiphospholipid syndrome.

Ultraspecific electrochemical DNA biosensor by coupling spontaneous cascade DNA branch migration and dual-signaling sensing strategy.

Using spontaneous cascade DNA branch migration and dual-signaling sensing strategy, we developed a novel universal electrochemical biosensor for the highly specific and sensitive detection of nucleic acids. A target strand (Ts) competitively hybridized with a ferrocene (Fc)-labeled signal probe (Fc-S1), which was blocked by a protector strand (Ps), after strand displacement to form the Ts/Fc-S1 duplex. A methylene blue (MB)-modified signal probe (MB-S2) was immobilized on the Au electrode surface by hybridizing with a thiolated capture probe (Cp). Then, the obtained reactants (Ts/Fc-S1 and MB-S2/Cp) suffered spontaneous DNA branch migration and produced two hybridization products (Fc-S1/Cp and MB-S2/Ts). These reactions led to the dissociation of MB molecules and the collection of Fc molecules. The detection mechanism of this DNA biosensor involved distance variation between the redox tags and the Au electrode, which was associated with target-induced cascade DNA branch migration. Moreover, we rationally designed the cascade DNA branch migration to occur spontaneously with ΔG° ≈ 0, at which slight thermodynamic changes caused by base mismatch exerted a disproportionately large effect on the hybridization yield. This "signal-on/off" sensing system exhibited a remarkable analytical performance and an ultrahigh discrimination capability even against a single-base mismatch. The maximum discrimination factor (DF) of base mutations or alterations can reach 17.9. Therefore, our electrochemical biosensor might hold a great potential for further applications in biomedical research and early clinical diagnosis.

DNA-AuNPs based signal amplification for highly sensitive detection of DNA methylation, methyltransferase activity and inhibitor screening.

A sensitive and selective electrochemical method was developed for the detection of DNA methylation, determination of DNA methyltransferase (MTase) activity and screening of MTase inhibitor. Methylene blue (MB) was employed as electrochemical indicator and DNA-modified gold nanoparticles (AuNPs) were used as signal amplification unit because the DNA strands in this composite have strong adsorption ability for MB. First, the thiolated single-stranded DNA S1 was self-assembled on gold electrode, hybridization between the lower portion of DNA S1 and its complementary DNA S2 formed an identical double-stranded tetranucleotide target sequence for both DNA adenine methylation (Dam) MTase and methylation-resistant endonuclease Mbo I, then the upper portion of DNA S1 was hybridized with its complementary DNA S3 modified on AuNPs to bring the DNA S3-AuNPs amplification units onto the electrode. The DNA S1/S2/S3-AuNPs bioconjugate has lots of DNA strands, and they can adsorb abundant MB. Mbo I endounuclease could not cleave the identical target sequence after it was methylated by Dam MTase. On the contrary, the sequence without methylation could be cleaved, which would decrease the amount of adsorbed MB. The presence of redox-active MB was detected electrochemically by differential pulse voltammetry (DPV). Thus, the activity of Dam MTase and methylation status were sensitively converted to the DNA S3-AuNPs amplified DPV signals. The DPV signal demonstrated a linear relationship with logarithm of Dam concentration ranging from 0.075 to 30U/mL, achieving a detection limit of 0.02U/mL (S/N=3). Also, screening of Dam MTase inhibitor 5-fluorouracil was successfully investigated using this fabricated sensor.

One-step fabrication of integrated disposable biosensor based on ADH/NAD+/meldola's blue/graphitized mesoporous carbons/chitosan nanobiocomposite for ethanol detection.

A novel strategy to simplify the dehydrogenase-based electrochemical biosensor fabrication through one-step drop-coating nanobiocomposite on a screen printed electrode (SPE) was developed. The nanobiocomposite was prepared by successively adding graphitized mesoporous carbons (GMCs), meldola's blue (MDB), alcohol dehydrogenase (ADH) and cofactor nicotinamide adenine dinucleotide (NAD(+)) in chitosan (CS) solution. MDB/GMCs/CS film was prepared. Cyclic voltammetry measurements demonstrated that MDB was strongly adsorbed on GMCs. After optimizing the concentration of MDB and the working potential, the MDB/GMCs/CS film presented a fast amperometric response (5s), excellent sensitivity (10.36 nA µM(-1)), wide linear range (10-410 µM) toward NADH and without any other interference signals (such as AA, UA, DA, H2O2 and metal ions). Furthermore, concentrations of ADH and NAD(+) in nanobiocomposite and the detection conditions (temperature and pH) were also optimized. The constructed disposable ethanol biosensor showed an excellent linear response ranged from 0.5 to 15 mM with high sensitivity (67.28 nA mM(-1)) and a low limit of detection (80 µM) and a remarkable long-term stability (40 days). The intra-batch and inter-batch variation coefficients were both less than 5% (n=5). The ethanol recovery test demonstrated that the proposed biosensor offered a remarkable and accurate method for ethanol detection in the real blood samples.

Three-dimensional cultures of human endometrial cells on Matrigel mimic in vivo morphology.

BACKGROUND: The regulation of endometrial physiology and morphogenesis by the paracrine effectors has been well established using in vivo studies. A more complete understanding of the endometrial function has been delayed due, in part, to a lack of appropriate culture models. In this study, we aimed to simulate the in vivo three-dimensional (3-D) growth pattern of endometrial cells using a 3-D in vitro culture system. METHODS: Isolated endometrial epithelial cells, stromal cells and RL95-2 cells were seeded into culture chambers coated with the extracellular matrix Matrigel and observed using light microscopy. Fluorescence staining and immunohistochemistry were used to assess the morphology. RESULTS: Depending on the culture conditions, epithelial cells and RL95-2 cells formed multicellular structures on Matrigel; stromal cells remained individually distinguishable or grew together to form 3-D lattice-like structures. CONCLUSIONS: Matrigel provided a good microenvironment for culturing endometrial cells. The cells cultured in the Matrigel-coated chambers closely resembled those seen in vivo.