Enhancing CRISPR/Cas-mediated detection of nucleic acids using PNIPAM-based reporters.

Currently most conventional reporters in CRISPR/Cas system, including fluorophore-quencher (FQ) and magnetic bead (MB)-based reporters, encounter limitations in terms of sensitivity and compatibility. To overcome these challenges, we developed novel reporters for CRISPR/Cas systems based on thermo-responsive poly(N-isopropylacrylamide) (PNIPAM). Below the lower critical solution temperature (LCST), PNIPAM-based reporters exhibited a liquid state and can be cleaved by Cas proteins in a homogeneous reaction, preserving function and structure of Cas proteins while effectively accelerating the reaction kinetics.

Methodological establishment and diagnostic value of a multiplex fluorescent PCR assay for the detection of three fastidious respiratory pathogens.

Background: Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis are common fastidious bacteria responsible for respiratory tract infections, particularly in children and immunocompromised individuals. Due to their demanding growth requirements, traditional culture methods often yield low sensitivity and delayed results, posing challenges for early and accurate diagnosis.

Objective: To establish a TaqMan probe-based multiplex fluorescent PCR method for the simultaneous rapid detection and identification of three important respiratory fastidious pathogens: Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis.

Assessment of the effect of routine-dose vitamin D3 supplementation in Chinese women of childbearing age with vitamin D insufficiency.

Background: It was reported that vitamin D deficiency is common in Chinese women of childbearing age. Although most government dietary guidelines recommend routine-dose (400-800 IU/d), the evidence from intervention studies using routine-dose is quite limited.

Objective: In this study, the effects of daily supplementation with routine-dose of vitamin D3 in women of childbearing age with vitamin D insufficiency on vitamin D nutritional status and bone metabolism markers were assessed by dynamic tracking, and the thresholds for appropriate levels of vitamin D in this population was also explored.

Development and performance evaluation of a multiplex fluorescent PCR method for the detection of fastidious pathogens causing respiratory infections in children.

Introduction: This study was performed to develop a multiplex fluorescent PCR method for the concurrent detection of six fastidious bacteria associated with respiratory tract infections. These bacteria include Streptococcus pneumoniae（SPN）, Bordetella pertussis (BP), Neisseria meningitidis (NM), Legionella pneumophila (LP), Moraxella catarrhalis (MC), and Haemophilus influenzae (HI).

Methodology: A multiplex fluorescent PCR test using SYBR Green as a DNA dye was developed and optimised.

Epidemiological characteristics and antimicrobial resistance of extensively drug-resistant in ICU wards.

is a significant nosocomial pathogen, particularly problematic due to its extensive drug resistance. This study investigates 56 extensively drug-resistant (XDRAB) strains collected from various ICU wards at Jinhua Central Hospital, Zhejiang Province, China. Strains were isolated from diverse clinical samples, including sputum, blood, cerebrospinal fluid, and wound secretions.

[Effects of serum vitamin K levels in early pregnancy on pregnancy-related diseases and adverse pregnancy outcomes].

Objective: To explore the relationship between vitamin K levels and pregnancy-related diseases and adverse pregnancy outcomes in early pregnancy.

Methods: A convenience sampling method was used to select 367 women in early pregnancy who were enrolled in the Shunyi Women's and Children's Hospital of Beijing Children's Hospital from October 2018 to September 2019. Basic information such as age, height, and pre-pregnancy weight of the respondents was collected, blood samples were taken and the concentration of vitamin K(vitamin K1, MK-4 and MK-7) was measured by liquid chromatography tandem mass spectrometry(LC-MS/MS).

Highly Sensitive Digital and Analog Immunoassay Based on Submicron Magnetic Beads and Fluorescent Microspheres.

The detection of disease-related protein biomarkers plays a crucial role in the early diagnosis, treatment, and monitoring of diseases. The concentrations of protein biomarkers can vary significantly in different diseases or stages of the same disease. However, most of the existing analytical methods cannot simultaneously meet the requirements of high sensitivity and a wide dynamic range.

Quantum Dot Reporters Designed for CRISPR-Based Detection of Viral Nucleic Acids.

Diagnostic methods based on CRISPR technology have shown great potential due to their highly specific, efficient, and sensitive detection capabilities. Although the majority of the current studies rely on fluorescent dye-quencher reporters, the limitations of fluorescent dyes, such as poor photostability and small Stokes shifts, urgently necessitate the optimization of reporters. In this study, we developed innovative quantum dot (QD) reporters for the CRISPR/Cas systems, which not only leveraged the advantages of high photoluminescence quantum yield and large Stokes shifts of QDs but were also easily synthesized through a simple one-step hydrothermal method.

Ultra-sensitive detection of SARS-CoV-2 S1 protein by coupling rolling circle amplification with poly(N-isopropylacrylamide)-based sandwich-type assay.

In the past few years, the COVID-19 pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) seriously threatens global public health security due to its high contagiousness. It remains of vital importance to develop a rapid and sensitive assay for SARS-CoV-2. In this work, we proposed a sandwich-type assay based on poly(N-isopropylacrylamide) (PNIPAM), allowing efficient detection of the SARS-CoV-2 S1 protein in the homogeneous solution.

Study on the Dose-Response Relationship between Magnesium and Type 2 Diabetes of Childbearing Women in the China Adult Chronic Disease and Nutrition Surveillance 2015.

: Magnesium (Mg) is an essential element and participates in many metabolic pathways. Many studies have found a certain negative correlation between magnesium and blood glucose parameters, but the dose-response relationship between them is still a relatively narrow research field. We aim to explore the dose-response relationship between plasma and dietary Mg and type 2 diabetes (T2DM) among childbearing women in a nationally representative sample.

Rare case of septic shock combined with meningitis caused by Pasteurella multocida without a history of cat and dog bites.

Background: Pasteurella multocida is a zoonotic pathogen that mainly causes local skin and soft tissue infections in the human body through cat and dog bites. It rarely causes bacteraemia (or sepsis) and meningitis. We reported a case of septic shock and meningitis caused by P.

A Facile Strategy for Multiplex Protein Detection by a Fluorescent Microsphere-Based Digital Immunoassay.

The digital immunoassay is a highly sensitive detection technique based on single-molecule counting and is widely used in the ultrasensitive detection of biomarkers. Herein, we developed a fluorescent microsphere-based digital immunoassay (FMDIA) by employing fluorescent microspheres as both the carriers for immunoreaction and fluorescent reports for imaging. In this approach, the target protein in the sample was captured by fluorescent microspheres to form a biotin-labeled sandwich immunocomplex, and then, the fluorescent microspheres containing the target protein molecules were captured by adding streptavidin-coated magnetic beads (SA-MBs).

Amplification-free detection of Mpox virus DNA using Cas12a and multiple crRNAs.

Mpox virus (MPXV) is a zoonotic DNA virus that caused human Mpox, leading to the 2022 global outbreak. MPXV infections can cause a number of clinical syndromes, which increases public health threats. Therefore, it is necessary to develop an effective and reliable method for infection prevention and control of epidemic.

[Relationship between vitamin D and parathyroid hormone in Chinese elderly people].

Objective: To explore the relationship between serum 25-hydroxyvitamin D(25(OH)D) and serum parathyroid hormone(PTH) level in Chinese people aged 50 years and above, and to probe the optimum threshold for vitamin D sufficiency preliminarily, and apply this threshold to predict the risk of metabolic syndrome(Mets) in this population.

Methods: A total of 750 serum samples were selected from the biological samples' bank of Chinese Chronic Diseases and Nutritional Survey(CCDNS, 2015-2017) by stratified sampling, basic information(including age, gender, season, etc. ) were collected from questionnaire and physical measurement of the subjects were conducted unified.

Nucleic Acid Amplification-Free Digital Detection Method for SARS-CoV-2 RNA Based on Droplet Microfluidics and CRISPR-Cas13a.

Most of the methods currently developed for RNA detection based on CRISPR were combined with nucleic acid amplification. As a result, such methods inevitably led to certain disadvantages such as multiple operations, expensive reagents, and amplification bias. To solve the above problems, we developed a highly sensitive and specific nucleic acid amplification-free digital detection method for SARS-CoV-2 RNA based on droplet microfluidics and CRISPR-Cas13a.

Three-stage pattern of rapid increase, plateau, and subsequent decline in vitamin D concentration during pregnancy among Chinese women: a large-scale survey.

Background: There is an incomplete understanding of fluctuations in vitamin D (VitD) concentration during pregnancy among Chinese women. Furthermore, previous research has yielded conflicting results in this area. This study aims to investigate the changes in VitD status and deficiency in Chinese pregnant women across various age groups, gestational weeks, and as well as seasonal variations through conducting a large-scale survey.

Free and bioavailable 25-hydroxyvitamin D thresholds for bone metabolism and their associations with metabolic syndrome in Chinese women of childbearing age.

Objective: The free hormone hypothesis suggests that free and bioavailable 25-hydroxyvitamin D [25(OH)D] may better reflect vitamin D bioactivity. This study aimed to determine the free and bioavailable 25(OH)D characteristics, estimate their thresholds based on parathyroid hormone (PTH) and bone turnover markers (BTMs), assess their associations with the risk of metabolic syndrome (MetS), and evaluate their potential advantages.

Methods: A cross-sectional study was conducted using a nationally representative database ( = 1,505, female, 18-45 years).

MicroRNA-32-3p facilitates cerebral ischemia/reperfusion injury through inhibiting Cab39/AMPK.

Oxidative stress is a key pathogenic factor of cerebral ischemia/reperfusion (I/R) injury. MicroRNA-32-3p (miR-32-3p) plays critical roles in regulating ischemic diseases; however, its role in oxidative stress and cerebral I/R injury remains elusive. Primary cortical neurons and rats were treated with the agomir, antagomir and matched controls of miR-32-3p, and then received oxygen glucose deprivation/reperfusion (OGD/R) or I/R stimulation.

Association of Body Iron Metabolism with Type 2 Diabetes Mellitus in Chinese Women of Childbearing Age: Results from the China Adult Chronic Disease and Nutrition Surveillance (2015).

High iron stores have been reported to be associated with type 2 diabetes mellitus (T2DM). However, evidence for the associations of iron metabolism with T2DM is inconsistent, and whether there is a threshold effect remains controversial. In the present study, we aimed to examine the associations between various iron biomarkers and the risk of T2DM as well as impaired glucose metabolism (IGM) and hyperglycemia in Chinese women of childbearing age.

The Magnesium Status and Suggested Reference Ranges of Plasma Magnesium, Calcium, and Calcium/Magnesium Ratio in Chinese Adults over 45 Years Old.

Magnesium (Mg) is an essential nutrient that participates in various enzymatic reactions and regulates important biological functions. The distribution and reference ranges in China have not been reported in populations more than 45 years old. This study aimed to assess the magnesium status and determine the reference values of plasma Mg, Ca, and Ca/Mg ratios for China's population more than 45 years old.

CRISPR/Cas13a Trans-Cleavage-Triggered Catalytic Hairpin Assembly Assay for Specific and Ultrasensitive SARS-CoV-2 RNA Detection.

New coronavirus (SARS-CoV-2), which has caused the coronavirus disease 2019 (COVID-19) pandemic, has brought about a huge burden on global healthcare systems. Rapid and early detection is important to prevent the spread of the pandemic. Here, an assay based on CRISPR/Cas13a and catalytic hairpin assembly (CHA), termed as Cas-CHA, was developed for ultrasensitive and specific detection of SARS-CoV-2 RNA.

[Association of vitamin D gene polymorphisms and serum 25-hydroxyvitamin D in Chinese women of childbearing age].

Objective: To analyze the relationship between vitamin D(VitD)-related single nucleotide polymorphism(SNP) and 25-hydroxyvitamin D(25(OH) D) levels and VitD nutritional status.

Methods: A total of 1507 women of childbearing age aged 18-45 were selected from the sample bank of "2015 Chinese adult chronic disease and nutrition monitoring". Basic information(including region, season, age, height, weight, etc.

The Association of Dietary Magnesium and Prediabetes in Childbearing Chinese Women: Results from China Nutrition and Health Surveillance (2015-2017).

Although several studies have shown the relationship between magnesium and diabetes, there are few studies regarding the association between magnesium status and prediabetes. This study aimed to examine the association between dietary magnesium intake and the risk of prediabetes in childbearing Chinese women (18−44 years). A total of 1981 participants without diabetes were randomly selected from China Nutrition and Health Surveillance (CNHS) in 2015 by considering the regional types and monitoring points, including 1420 normal fasting glucose (NFG) people and 561 prediabetes cases.