Standardization and application of ARMS TaqMan real-time PCR for screening of folate metabolism genes in Han Chinese.

Folate has antioxidant properties, and low concentration in seminal plasma may be associated with increased DNA damage in sperm. Mutations of the methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR) genes, including MTHFR C677T (rs1801133), MTHFR A1298C (rs1801131), and MTRR A66G (rs1801394), can lead to decreased activity of the encoded folate metabolic enzymes, thereby affecting male reproduction. The current SNP detection methods commonly used in clinical practice have some shortcomings, such as long time-consuming, complex detection steps, or high cost. The purpose of this study was to establish a simple, time-saving, sensitive, accurate, and easy to clinical popularization method for folate metabolism gene detection. We combined ARMS-PCR with TaqMan fluorescent probe to establish an ARMS TaqMan real-time PCR detection method. According to the variation of rs1801131, rs1801133, and rs1801394, two specific primers (one wild type and one mutant) were designed. Mismatched nucleotides were introduced at the penultimate or third position to improve the specificity of the primer. Specific TaqMan probe was introduced to detect PCR products to improve the sensitivity of the method. The results showed that the sensitivity of ARMS TaqMan real-time PCR in SNP genotyping was 1 ng, and the accuracy was 100%. A total of 249 clinical samples were detected by the established method, and the correlation between three SNPs and semen quality was analyzed. We found that individuals carrying the AG + GG genotype of rs1801394 had a lower risk of abnormal semen quality. In conclusion, we developed a highly sensitive, accurate, rapid, and easy to be popularized method for detecting SNPs of rs1801394, rs1801131, and rs1801133. ARMS TaqMan real-time PCR is a reliable SNP genotyping method in folate metabolism genes.

Association of SNPs in zinc transporter genes on seminal plasma zinc levels in humans.

This study is to examine the effects of single nucleotide polymorphisms (SNPs) of SLC30A and SLC39A on seminal plasma zinc concentration. Blood and seminal plasma samples were collected from outpatients. SNPs of zinc transporters were analyzed by next Generation sequencing technology, and seminal plasma zinc concentration were determined by inductively coupled plasma optical emission spectrometry. Our date showed nine SNPs (SLC30A8 rs2466295, rs2466294, SLC30A10 c.-160 C>G, SLC39A8 rs9331, rs9705, rs151392, rs151393, SLC39A11 rs9912126, SLC39A14 rs1051708) were significantly associated with seminal plasma zinc concentration, and 14 SNPs (SLC30A8 rs2466295, rs2466294, SLC30A10 c.-160 C>G, SLC39A6 rs148550301, SLC39A8 rs9331, rs9705, rs151392, rs151393, SLC39A11 rs9912126, rs61736066, rs36041371 and SLC39A14 rs1051708, rs76963096, rs17060854) were found to be significantly associated with total zinc per ejaculate. The seminal plasma zinc concentrations and total zinc per ejaculate were associated with the number of SNPs, and decreased significantly when five SNPs (SLC39A8 rs9331, rs9705, rs151392, rs151393 and SLC39A14 rs1051708) were a combination of homozygous genotype. Our findings suggest that different zinc transporter SNPs may significantly affect seminal plasma zinc levels.

Differentially expressed microRNA in testicular tissues of hyperuricaemia rats.

This study is to identify the differentially expressed miRNAs in testicular tissues of rats with hyperuricaemia-induced male infertility. We found that the hyperuricaemia model group had significantly increased serum uric acid, while significantly decreased sperm concentration and motile sperm percentage than normal group (p < .05). A total of 39 differentially expressed miRNAs were identified in the testicular tissues of hyperuricaemia rats compared with the control rats, ten of which were validated by real-time PCR. The target mRNAs of 7 differentially expressed miRNAs (miR-10b-5p, miR-26a-5p, miR-136-5p, miR-151-3p, miR-183-5p, miR-362-3p and miR-509-5p) from 3'-untranslated region binding perspective were enriched in signalling pathways of Wnt, Jak-STAT, mTOR and MAPK. The target mRNAs of 6 differentially expressed miRNAs (miR-136-5p, miR-144-3p, miR-99a-5p, miR-509-5p, miR-451-5p and miR-362-3p) from coding sequence binding perspective were enriched in signalling pathways of Calcium, Notch and MAPK. The functions of miRNAs in testicular tissues of rats with hyperuricaemia were revealed by the differentially expressed miRNAs (miR-183-5p, miR-99a-5p, miR-10b-5p, miR-151-3p, miR-26a-5p, miR-451-5p, miR-362-3p, miR-136-5p, miR-144-3p and miR-509-5p)-mRNAs interaction network. The differentially expressed miRNAs in the testicular tissues of hyperuricaemia rats might shed light on the mechanism of hyperuricaemia-induced male infertility.

Prototype development of a three-stage, high-precision, low-jitter, wide-range digital delay generator fully utilizing on-chip resources.

This paper introduces the design and implementation of a prototype Digital Delay Generator (DDG) characterized by high precision, low jitter, and a wide delay range, fully realized within a Field Programmable Gate Array (FPGA). The DDG's architecture is based on an innovative combination of an embedded time-to-digital converter (TDC) and Multi-stage Time Interpolation (MTI) delay logic. The paper explores the factors influencing delay jitter during external trigger mode and carefully selects the optimal design approach for each element. The embedded TDC, which undergoes automatic calibration, accurately measures the time difference between the arrival of an external trigger and the FPGA's internal clock signal. When paired with the MTI delay logic, this allows for highly precise control over delay durations. A key aspect of this design is its sole dependence on the FPGA's built-in resources, ensuring simplicity in implementation and adaptability to various applications. Evaluation of the prototype has shown promising results, demonstrating a delay resolution as fine as 20 ps and maintaining a low jitter of 105 ps peak-to-peak (20 ps rms) when operated in the externally triggered mode.

Specific discrimination and efficient elimination of gram-positive bacteria by an aggregation-induced emission-active ruthenium (II) photosensitizer.

The infections caused by Gram-positive bacteria (G+) have seriously endangered public heath due to their high morbidity and mortality. Therefore, it is urgent to develop a multifunctional system for selective recognition, imaging and efficient eradication of G+. Aggregation-induced emission materials have shown great promise for microbial detection and antimicrobial therapy. In this paper, a multifunctional ruthenium (II) polypyridine complex Ru2 with aggregation-induced emission (AIE) characteristic, was developed and used for selective discrimination and efficient extermination of G+ from other bacteria with unique selectivity. The selective G+ recognition benefited from the interaction between lipoteichoic acids (LTA) and Ru2. Accumulation of Ru2 on the G+ membrane turned on its AIE luminescence and allowed specific G+ staining. Meanwhile, Ru2 under light irradiation also possessed robust antibacterial activity for G+in vitro and in vivo antibacterial experiments. To the best of our knowledge, Ru2 is the first Ru-based AIEgen photosensitizer for simultaneous dual applications of G+ detection and treatment, and inspires the development of promising antibacterial agents in the future.

Two tales of platform regimes in China's food-delivery platform economy.

This article brings the often-overlooked concept of the labor regime back to the study of China's food-delivery platform workers. Two tales of platform regimes emerge: individualized platform despotism and bureaucratized platform despotism, which apply to crowdsourcing couriers and dedicated delivery couriers, respectively. This study compares these two types of platform regimes in terms of their institutional foundation and labor organization. Despite different institutional arrangements and labor organization, both types of food-delivery couriers belong to a despotic platform regime revealing workers' subordination to the platform. In conclusion, it discusses the implications and limitations of this study.

A rapid "on-off-on" mitochondria-targeted phosphorescent probe for selective and consecutive detection of Cu2+ and cysteine in live cells and zebrafish.

The detection of mitochondrial Cu2+ and cysteine is very important for investigating cellular functions or dysfunctions. In this study, we designed a novel cyclometalated iridium(iii) luminescence chemosensor Ir bearing a bidentate chelating pyrazolyl-pyridine ligand as a copper-specific receptor. The biocompatible and photostable Ir complex exhibited not only mitochondria-targeting properties but also an "on-off-on" type phosphorescence change for the reversible dual detection of Cu2+ and cysteine. Ir had a highly sensitive (detection limit = 20 nM) and selective sensor performance for Cu2+ in aqueous solution due to the formation of a non-phosphorescent Ir-Cu(ii) ensemble through 1 : 1 binding. According to the displacement approach, Ir was released from the Ir-Cu(ii) ensemble accompanied with "turn-on" phosphorescence in the presence of 0-10 µM cysteine, with a low detection limit of 54 nM. This "on-off-on" process could be accomplished within 30 s and repeated at least five times without significant loss of signal strength. Moreover, benefiting from its good permeability, low cytotoxicity, high efficiency, and anti-interference properties, Ir was found to be suitable for imaging and detecting mitochondrial Cu2+ and cysteine in living cells and zebrafish.