The present study aimed to explore whether hydrogen sulfide (H₂S) improved hypoxic pulmonary hypertension (HPH) in rats by inhibiting aerobic glycolysis-pyroptosis. Male Sprague-Dawley (SD) rats were randomly divided into normal group, normal+NaHS group, hypoxia group, and hypoxia+NaHS group, with 6 rats in each group. The control group rats were placed in a normoxic (21% O₂) environment and received daily intraperitoneal injections of an equal volume of normal saline. The normal+NaHS group rats were placed in a normoxic environment and intraperitoneally injected with 14 μmol/kg NaHS daily. The hypoxia group rats were placed in a hypoxia chamber, and the oxygen controller inside the chamber maintained the oxygen concentration at 9% to 10% by controlling the N₂ flow rate. An equal volume of normal saline was injected intraperitoneally every day. The hypoxia+NaHS group rats were also placed in an hypoxia chamber and intraperitoneally injected with 14 μmol/kg NaHS daily. After the completion of the four-week modeling, the mean pulmonary artery pressure (mPAP) of each group was measured using right heart catheterization technique, and the right ventricular hypertrophy index (RVHI) was weighed and calculated. HE staining was used to observe pathological changes in lung tissue, Masson staining was used to observe fibrosis of lung tissue, and Western blot was used to detect protein expression levels of hexokinase 2 (HK2), pyruvate dehydrogenase (PDH), pyruvate kinase isozyme type M2 (PKM2), nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), GSDMD-N-terminal domain (GSDMD-N), Caspase-1, interleukin-1β (IL-1β) and IL-18 in lung tissue. ELISA was used to detect contents of IL-1β and IL-18 in lung tissue. The results showed that, compared with the normal control group, there were no significant changes in all indexes in the normal+NaHS group, while the hypoxia group exhibited significantly increased mPAP and RVHI, thickened pulmonary vascular wall, narrowed lumen, increased collagen fibers, up-regulated expression levels of aerobic glycolysis-related proteins (HK2 and PKM2), up-regulated expression levels of pyroptosis-related proteins (NLRP3, GSDMD-N, Caspase-1, IL-1β, and IL-18), and increased contents of IL-1β and IL-18. These changes of the above indexes in the hypoxia group were significantly reversed by NaHS. These results suggest that H₂S can improve rat HPH by inhibiting aerobic glycolysis-pyroptosis.
Animals ; Rats, Sprague-Dawley ; Male ; Hypertension, Pulmonary/metabolism* ; Glycolysis/drug effects* ; Hydrogen Sulfide/therapeutic use* ; Hypoxia/complications* ; Rats ; Pyroptosis/drug effects*

The aim of this study was to investigate the contribution of lung zinc ions to pathogenesis of lung ischemia/reperfusion (I/R) injury in rats. Male Sprague Dawley (SD) rats were randomly divided into control group, lung I/R group (I/R group), lung I/R + low-dose zinc chloride group (LZnCl₂+I/R group), lung I/R + high-dose ZnCl₂ group (HZnCl₂+I/R group), lung I/R + medium-dose ZnCl₂ group (MZnCl₂+I/R group) and TPEN+MZnCl₂+I/R group (n = 8 in each group). Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure the concentration of zinc ions in lung tissue. The degree of lung tissue injury was analyzed by observing HE staining, alveolar damage index, lung wet/dry weight ratio and lung tissue gross changes. TUNEL staining was used to detect cellular apoptosis in lung tissue. Western blot and RT-qPCR were used to determine the protein expression levels of caspase-3 and ZIP8, as well as the mRNA expression levels of zinc transporters (ZIP, ZNT) in lung tissue. The mitochondrial membrane potential (MMP) of lung tissue was detected by JC-1 MMP detection kit. The results showed that, compared with the control group, the lung tissue damage, lung wet/dry weight ratio and alveolar damage index were significantly increased in the I/R group. And in the lung tissue, the concentration of Zn²⁺ was markedly decreased, while the cleaved caspase-3/caspase-3 ratio and apoptotic levels were significantly increased. The expression levels of ZIP8 mRNA and protein were down-regulated significantly, while the mRNA expression of other zinc transporters remained unchanged. There was also a significant decrease in MMP. Compared with the I/R group, both MZnCl₂+I/R group and HZnCl₂+I/R group exhibited significantly reduced lung tissue injury, lung wet/dry weight ratio and alveolar damage index, increased Zn²⁺ concentration, decreased ratio of cleaved caspase-3/caspase-3 and apoptosis, and up-regulated expression levels of ZIP8 mRNA and protein. In addition, the MMP was significantly increased in the lung tissue. Zn²⁺ chelating agent TPEN reversed the above-mentioned protective effects of medium-dose ZnCl₂ on the lung tissue in the I/R group. The aforementioned results suggest that exogenous administration of ZnCl₂ can improve lung I/R injury in rats.
Animals ; Reperfusion Injury/pathology* ; Male ; Rats, Sprague-Dawley ; Rats ; Chlorides/administration & dosage* ; Lung/pathology* ; Zinc Compounds/administration & dosage* ; Apoptosis/drug effects* ; Caspase 3/metabolism* ; Cation Transport Proteins/metabolism*

Based on whole-genome identification of the TPS gene family in Perilla frutescens and screening, cloning, bioinformatics, and expression analysis of the synthetic enzyme for the insect-resistant component germacrene D, this study lays the foundation for understanding the biological function of the TPS gene family and the insect resistance mechanism in P. frutescens. This study used bioinformatics tools to identify the TPS gene family of P. frutescens based on its whole genome and predicted the physicochemical properties, systematic classification, and promoter cis-elements of the proteins. The relative content of germacrene D was detected in both normal and insect-infested leaves of P. frutescens, and the germacrene D synthase was screened and isolated. Gene cloning, bioinformatics analysis, and expression profiling were then performed. The results showed that a total of 99 TPS genes were identified in the genome, which were classified into the TPS-a, TPS-b, TPS-c, TPS-e/f, and TPS-g subfamilies. Conserved motif analysis showed that the TPS in P. frutescens has conserved structural characteristics within the same subfamily. Promoter cis-element analysis predicted the presence of light-responsive elements, multiple hormone-responsive elements, and stress-responsive elements in the TPS family of P. frutescens. Transcriptome data revealed that most of the TPS genes in P. frutescens were highly expressed in the leaves. GC-MS analysis showed that the relative content of germacrene D significantly increased in insect-damaged leaves, suggesting that it may act as an insect-resistant component. The germacrene D synthase gene was screened through homologous protein binding gene expression and was found to belong to the TPS-a subfamily, encoding a 64.89 kDa protein. This protein was hydrophilic, lacked a transmembrane structure and signal peptide, and was predominantly expressed in leaves, with significantly higher expression in insect-damaged leaves compared to normal leaves. In vitro expression results showed that germacrene D synthase tended to form inclusion bodies. Molecular docking showed that farnesyl pyrophosphate(FPP) fell into the active pocket of the protein and interacted strongly with six active sites. This study provides a foundation for further research on the biological functions of the TPS gene family in P. frutescens and the molecular mechanisms underlying its insect resistance.
Perilla frutescens/chemistry* ; Plant Proteins/chemistry* ; Multigene Family ; Sesquiterpenes, Germacrane/metabolism* ; Alkyl and Aryl Transferases/chemistry* ; Phylogeny ; Gene Expression Regulation, Plant

Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

[This corrects the article DOI: 10.1016/j.apsb.2019.01.013.].

BACKGROUND:At present,the traditional powder sintering method is easy to introduce impurities in the process of preparing porous titanium,and the manufacturing of porous titanium still faces two major problems:impurity pollution and difficult control of the material forming process. OBJECTIVE:To prepare pure porous titanium with certain porosity,and analyze the microstructure evolution and properties of the porous titanium. METHODS:Porous titanium was prepared at a low energy density by selective laser melting technology.The parameter range of porous titanium with large porosity was obtained by measuring the porosity of the formed specimen,and the evolution of the microstructure and mechanical properties of the specimen in the range were analyzed. RESULTS AND CONCLUSION:(1)With the increase in energy density,the porosity of the porous titanium specimen decreased gradually.When the energy density was between 10.61 and 27.78 J/mm3,porous titanium with a porosity of 11.23%-33.67%could be formed.When the energy density was between 27.78-37.88 J/mm3,the forming parts were relatively dense.(2)The phase composition of porous titanium formed was mainly α titanium.With the increase in energy density,the porosity gradually decreased,and the pore morphology changed from irregularly connected pores to closed nearly spherical pores.The powder particles changed from a slightly sintered neck to a continuous fuse.The CT scan results revealed that there were a large number of connected pores in the forming specimen with a large specific surface area and the pore radius was roughly distributed between 2-6 μm at the energy density of 10.61 J/mm3.Simultaneously,porous titanium with compressive strength of 188-1 000 MPa could be obtained at the energy density of 10.61-27.78 J/mm3,which could meet the requirements of biomedical applications.(3)These results have confirmed that the selective laser melting technology can overcome the problems of impurity pollution and long manufacturing cycle caused by the traditional preparation process,and provide an effective solution for the preparation of porous titanium with excellent mechanical properties.

ObjectiveTo investigate the contamination status of Staphylococcus aureus in food and the presence of enterotoxin genes in Jiading District， Shanghai， and to provide a basis for the prevention and treatment of foodborne Staphylococcus aureus disease. MethodsFrom 2021 to 2023， 15 types of food were sampled for S. aureus testing， and the presence of five enterotoxin genes， including sea⁃see， was tested in the strains. ResultsOut of 705 food samples， 88 （12.48%） were positive for S. aureus. S. aureus was detected in 12 of the 15 food types， with the three food types with the highest positive rates being cold noodles （45.00%）， raw poultry （26.25%）， and vegetable salads （20.00%）. The enterotoxin gene carriage rate was 32.95% in food strains. The carriage rates for sea， seb， and sec were 7.95%， 12.50%， and 14.77%， respectively. Neither sed nor see was detected. The detection rate of strains carrying two types of enterotoxin genes was 2.27%. The enterotoxin carriage rates in strains from vegetables， beverages， and raw meat were 57.14%， 40.00%， and 30.00%， respectively. ConclusionThe S. aureus detection rate in food in Jiading District is much higher than the national average. The enterotoxin gene carriage rates are high， with food strains carrying sea， seb， and sec， with sec being the most prevalent. There is a need to enhance monitoring of S. aureus and enterotoxins， especially in high-risk foods such as noodles， vegetables， and non-packaged beverages.

ObjectiveTo investigate the prevalence of Staphylococcus aureus in patients with diarrhea, and to analyze the genes carriage of enterotoxin in the strains of these patients in Jiading District, Shanghai. MethodsFrom 2021 to 2023, anal swabs of diarrhea outpatients from one sentinel hospital and nine community health service centers in different townships in Jiading District, Shanghai, were tested for Staphylococcus aureus, from which five enterotoxin virulence genes such as SEA, SEB, SEC, SED, and SEE were tested simultaneously. ResultsA total of 1 080 anal swabs were collected, 81 of which were tested positive for S. aureus, with a detection rate of 7.50%, and the detection rate of S. aureus was similar in patients with diarrhea from 2021‒2023. There was no statistically significant difference in detection rates between males and females (χ²=0.821, P=0.365). S. aureus detection rate was highest in infants and young children with diarrhea (29.51%), followed by 14.06% in the people aged between 4‒<31 years, and 2.99% in those aged ≥31 years. Significant differences were observed in the detection rate of S. aureus in the diarrhoeal patients from different townships of Jiading District(χ²=66.134,P<0.05). The carriage rates of the 5 enterotoxin genes, namely SEA, SEB, SEC, SED, SEE, were 13.58%, 14.81%, 11.11%, 7.41%, and 0, respectively. ConclusionThe prevalence of S. aureus among the patients with diarrhea in Jiading District is relatively stable but with distinct geographical patterns. Children and adolescents are high-risk groups. SEB were the dominant gene, followed by SEA.