Investigation of the Antibacterial Properties and Mode of Action of Compounds From Urtica dioica L.

In recent years, the issue of antimicrobial resistance has gained significant global attention. The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infection rates worldwide has seen a rapid rise, increasing from 1%-5% in the mid-1980s to 60%-70% at present. This alarming increase in MRSA infection poses a serious threat to public health globally. Consequently, it is crucial to explore and identify effective drug candidates for combating MRSA. We researched the antibacterial properties of Urtica dioica L. Modern techniques such as systematic solvent extraction, macroporous resin chromatography, and silica gel column chromatography were utilized to isolate and detect components of MRSA. The most potent antibacterial active components were screened using fungal staining (K-B staining) and 2,3,5-triphenyltetrazolium chloride staining (TTC staining). Based on pharmacological activity guidance, we isolated a total of nine compounds from this plant. They were vanillic acid (compound A), quercetin-3-O-glucopyranoside (compound B), ursolic acid (compound C), vanillin (compound D), salicyl alcohol (compound E), kaempferol (compound F), quercetin (compound G), quercetin-3-O-galactoside (compound H), and isorhamnetin (compound I). Isolated compounds A, D, and E have better anti-MRSA activity. It inhibits bacterial division and growth during the logarithmic growth period and acts as a bacterial inhibitor. Inhibition may be mediated by the disruption of the bacterial cell structure, leading to leakage of contents including sugars, nucleic acids, and proteins. It may also be mediated by regulating phosphorus metabolism and disrupting the bacterial cell membrane potential to affect cellular metabolism.

Characterization of vB_ValM_PVA8, a broad-host-range bacteriophage infecting Vibrio alginolyticus and Vibrio parahaemolyticus.

Phage therapy was taken as an alternative strategy to antibiotics in shrimp farming for the control of Vibrio species of Vibrio parahaemolyticus and Vibrio alginolyticus, which cause substantial mortality and significant economic losses. In this study, a new Vibrio phage vB_ValM_PVA8 (PVA8), which could efficiently infect pathogenic isolates of V. alginolyticus and V. parahaemolyticus, was isolated from sewage water and characterized by microbiological and in silico genomic analyses. The phage was characterized to be a member of the Straboviridae family with elongated head and contractile tail by transmission electron microscopy. Genome sequencing showed that PVA8 had a 246,348-bp double-stranded DNA genome with a G + C content of 42.6%. It harbored totally 388 putative open reading frames (ORFs), among them 92 (23.71%) assigned to functional genes. Up to 27 transfer RNA (tRNA) genes were found in the genome, and the genes for virulence, antibiotic resistance, and lysogeny were not detected. NCBI genomic blasting results and the phylogenetic analysis based on the sequences of the large terminase subunits and the DNA polymerase indicated that PVA8 shared considerable similarity with Vibrio phage V09 and bacteriophage KVP40. The phage had a latent period of 20 min and a burst size of 309 PFUs/infected cell with the host V. alginolyticus, and it was stable over a broad pH range (4.0-11.0) and a wide temperature span (-80°C to 60°C), respectively, which may benefit its feasibility for phage therapy. In addition, it had the minimum multiplicity of infection (MOI) of 0.0000001, which revealed its strong multiplication capacity. The shrimp cultivation lab trials demonstrated that PVA8 could be applied in treating pathogenic V. parahaemolyticus infection disease of shrimp with a survival rate of 88.89% comparing to that of 34.43% in the infected group, and the pond application trails confirmed that the implementation of PVA8 could rapidly yet effectively reduce the level of the Vibrio. Taken together, PVA8 may be potential to be explored as a promising biological agent for Vibrio control in aquaculture farming industry.

New developments in non-exosomal and exosomal ncRNAs in coronary artery disease.

Aim & methods: Non-exosomal and exosomal ncRNAs have been reported to be involved in the regulation of coronary artery disease (CAD). Therefore, to explore the biological effects of non-exosomal/exosomal ncRNAs in CAD, the authors searched for studies published in the last 3 years on these ncRNAs in CAD and summarized their functions and mechanisms. Results: The authors summarized 120 non-exosomal ncRNAs capable of regulating CAD progression. In clinical studies, 47 non-exosomal and nine exosomal ncRNAs were able to serve as biomarkers for the diagnosis of CAD. Conclusion: Non-exosomal/exosomal ncRNAs are not only able to serve as biomarkers for CAD diagnosis but can also regulate CAD progression through ceRNA mechanisms and are a potential target for early clinical intervention in CAD.

ncRNAs are increasingly found to play regulatory roles in coronary artery disease (CAD), and transcriptome studies offer greater advantages for controlling CAD at its source. Therefore, the authors conducted an accurate search and summary of studies on CAD and ncRNAs published in the past 3 years to analyze the main pathological mechanisms in CAD progression, aiming to provide a research basis for clinical treatment of CAD.

Integrated bioinformatics analysis for novel miRNAs markers and ceRNA network in diabetic retinopathy.

In order to seek a more outstanding diagnosis and treatment of diabetic retinopathy (DR), we predicted the miRNA biomarkers of DR and explored the pathological mechanism of DR through bioinformatics analysis. Method: Based on public omics data and databases, we investigated ncRNA (non-coding RNA) functions based on the ceRNA hypothesis. Result: Among differentially expressed miRNAs (DE-miRNAs), hsa-miR-1179, -4797-3p and -665 may be diagnosis biomarkers of DR. Functional enrichment analysis revealed differentially expressed mRNAs (DE-mRNAs) enriched in mitochondrial transport, cellular respiration and energy derivation. 18 tissue/organ-specific expressed genes, 10 hub genes and gene cluster modules were identified. The ceRNA networks lncRNA FBXL19-AS1/miR-378f/MRPL39 and lncRNA UBL7-AS1/miR-378f/MRPL39 might be potential RNA regulatory pathways in DR. Conclusion: Differentially expressed hsa-miR-1179, -4797-3p and -665 can be used as powerful markers for DR diagnosis, and the ceRNA network: lncRNA FBXL19-AS1/UBL7-AS1-miR-378f-MRPL39 may represent an important regulatory role in DR progression.

X-linked recessive Kallmann syndrome: A case report.

BACKGROUND: Kallmann syndrome (KS), also known as hypogonadotropic hypogonadism (HH) or olfactory-gonadal dysplasia, is a genetic condition in which the primary symptom is a failure to begin puberty or a failure to fully complete it. It occurs in both males and females and has the additional symptoms of hypogonadism and almost invariably infertility. The condition has a low prevalence that is estimated to be 1 in 4000 for male HH cases overall and 1:50000 for KS. It is three to five times more common in males than females. Whether this is a true sex imbalance or a reflection of how difficult KS/HH is to diagnose correctly in males vs females has yet to be fully established. CASE SUMMARY: This article reports a 26-year-old male presenting with delayed puberty. The synthetic decapeptide luteinizing hormone-releasing hormone stimulation test showed that the secretion levels of follicle-stimulating hormone and luteinizing hormone were delayed. The eigengenes commonly associated with idiopathic HH (IHH) were screened, and an X-linked recessive (KAL-1) mutation was found. His gonadotropin and testosterone levels increased significantly after pulsatile gonadotropin-releasing hormone (GnRH) subcutaneous therapy by pump. A relevant literature review on the recent advances in the diagnosis and treatment of KS and genetic counseling was conducted. CONCLUSION: KS is caused by a KAL-1 mutation that follows an X-linked recessive inheritance pattern. Pulsatile GnRH subcutaneous therapy by pump was effective in this patient.

Subclinical Hypothyroidism and Sperm DNA Fragmentation: A Cross-sectional Study of 5401 Men Seeking Infertility Care.

CONTEXT: Our previous study showed that paternal subclinical hypothyroidism (SCH) had a detrimental effect on the clinical outcomes of assisted reproductive technologies. However, it remains to be determined whether paternal SCH affects sperm DNA integrity. OBJECTIVE: To investigate the association between SCH and sperm DNA fragmentation in men seeking infertility care. METHODS: This cross-sectional study included 4983 men with euthyroidism and 418 men with SCH seeking infertility treatment in a tertiary care academic medical center between January 2017 and December 2021. The outcome measures were the absolute DNA fragmentation index (DFI) and the risk of abnormal DFI (defined as DFI ≥ 25% or ≥ 30%). RESULTS: The mean (SD) age of men with euthyroidism and men with SCH was 34.20 (5.97) and 35.35 (6.48) years, respectively (P < 0.001). The difference in DFI was not statistically significant (adjusted mean: 19.7% vs 18.9% in the SCH and euthyroidism groups, respectively; P = 0.07) after confounder adjustment. A DFI ≥25% was significantly more frequent in men with SCH (20.57%) than in men with euthyroidism (14.49%) after confounder adjustment [odds ratio (OR) 1.43 (95% CI 1.09-1.88)]. DFI ≥ 30% was also significantly more common in men with SCH (11.72%) than in men with euthyroidism [6.74%; OR 1.84 (95% CI 1.34-2.52)]. In addition, thyroid-stimulating hormone concentration was significantly associated with an increased risk of having a DFI ≥25% (P < 0.001) or ≥30% (P = 0.011). CONCLUSION: SCH was significantly associated with an increased risk of an abnormal DFI.

A Novel Bi-Functional Fibrinolytic Enzyme with Anticoagulant and Thrombolytic Activities from a Marine-Derived Fungus Aspergillus versicolor ZLH-1.

Fibrinolytic enzymes are important components in the treatment of thrombosis-associated disorders. A new bi-functional fibrinolytic enzyme, versiase, was identified from a marine-derived fungus Aspergillus versicolor ZLH-1. The enzyme was isolated from the fungal culture through precipitation with ammonium sulfate at 90% saturation. Additionally, it was further purified by DEAE-based ion-exchange chromatography, with a recovery of 20.4%. The fibrinolytic enzyme presented as one band on both SDS-PAGE and fibrin-zymogram, with a molecular mass of 37.3 kDa. It was elucidated as a member of metalloprotease in M35 family by proteomic approaches. The homology-modeling analysis revealed that versiase shares significant structural homology wuth the zinc metalloendopeptidase. The enzyme displayed maximum activity at 40 °C and pH 5.0. The activity of versiase was strongly inhibited by the metalloprotease inhibitors EDTA and BGTA. Furthermore, versiase hydrolyzed fibrin directly and indirectly via the activation of plasminogen, and it was able to hydrolyze the three chains (α, ß, γ) of fibrin(ogen). Additionally, versiase demonstrated promising thrombolytic and anticoagulant activities, without many side-effects noticed. In conclusion, versiase appears to be a potent fibrinolytic enzyme deserving further investigation.

Characterization and Genomic Analysis of a Bacteriophage with Potential in Lysing Vibrio alginolyticus

Vibrio alginolyticus is one of the major pathogens causing vibriosis to a variety of aquatic animals as well as bringing about severe food safety concerns. Nowadays, phage therapy has received increasing attention as an alternative to the antibiotics that have being limited for use in aquaculture industries. In this work, a potent bacteriophage, vB_ValM_PVA23 (PVA23), which efficiently infects pathogenic strains of V. alginolyticus, was isolated from sewage water and characterized by microbiological and genomic analyses. Based on the transmission electronic observation, the phage was characterized to be the Myoviridae family. It has a latent period of 10 min and a burst size of 203 PFUs/infected bacterium, and was stable over a broad pH range (5.0−11.0) and a wide temperature span (−80 °C to 60 °C), respectively. Genome sequencing results show that PVA23 has a 246,962-bp double-stranded DNA with a G + C content of 41.25%. The lab and plant shrimp farming trials demonstrated that phage preparation derived from PVA23 out-performed the chemical disinfectant iodine treatment in the prevention of V. alginolyticus propagation, and the phage application could rapidly yet significantly reduce the level of V. alginolyticus in the pond within 12 h, with negligible rebound observed. These results suggests that phage PVA23 has the potential to be used as an anti-V. alginolyticus agent in aquaculture industries.

Purification, characterization of Chondroitinase ABC from Sphingomonas paucimobilis and in vitro cardiocytoprotection of the enzymatically degraded CS-A.

An extracellular chondroitinase ABC (ChSase ABC) produced by Sphingomonas paucimobilis was purified to homogeneity through ammonium sulfate precipitation, DEAE-Sepharose Fast Flow and Sephadex G-100 chromatography. The molecular weight was 82.3 kDa. It showed specific lyase activity toward chondroitin sulfate A (CS-A), CS-B, CS-C and hyaluronan (HA). Using CS-A as substrate, the specific activity was 98.04 U/mg, the maximal reaction rate (Vmax) and Michaelis-Menten constant (Km) were 0.49 µmol/min/ml and 0.79 mg/ml, respectively. Highest activity was obtained at pH 6.5 and 40 °C, and Hg2+ could strongly inhibit the enzyme activity. Mass spectrometry analysis indicated CS-A was degraded to unsaturated disaccharides by ChSase ABC. In vitro cytotoxic tests showed that CS-A oligosaccharide at the concentration of 50 and 100 µg/ml could promote the proliferation of normal H9c2 myocardial cells, decrease the damage induced by isoproterenol (ISO) and accelerate the recovery of cells injured by ISO. These findings suggested that ChSase ABC from Sphingomonas paucimobilis could be a promising tool for the structural analysis and bioactive oligosaccharide preparation of glucosaminoglycans.

[Preliminary study on the aerobes distribution of nasal cavity from the healthy children and adults].

OBJECTIVE: To study whether there are differences in the bacteria distribution from the nasal cavity of the healthy children, teenagers and adults METHOD: The cotton swab specimens were taken from the nasal cavity of the healthy children, teenagers and adults for aerobic culture training. RESULT: Corynebacterium, coagulase-negative staphylococci, Staphylococcus aureus, Gram-negative Neisseria, alpha-hemolytic streptococcus and gram-negative bacillus with, six species of bacteria in total, were cultivated from the nasal cavity of 40 healthy children. The positive rate of bacterial culture was 80.0%; Corynebacterium, coagulase-negative staphylococci and Staphylococcus aureus were cultivated from the nasal cavity of 40 healthy teenagers and 56 healthy adults with the positive rate of bacterial culture was 90.0% from the healthy teenagers group and was 92.9% from the healthy adults group. CONCLUSION: There were obvious difference between the nasal cavity of the healthy children and adults with no obvious difference between the bacteria distribution from the nasal cavity of healthy teenagers and adults.

[Percutaneous dilational tracheostomy].

OBJECTIVE: To describe and introduce a new technique for percutaneous dilational tracheostomy without the aid of a bronchoscope. METHODS: Ten patients who underwent rotating dilation tracheostomy with percutwist set were prospectively studied. The time needed for the procedure, the grading of the difficulty, the amount of bleeding, and the complications for the procedure were evaluated. All procedures were performed without the aid of a bronchoscope. RESULTS: During the operations, blood pressure, ECG parameters were stable. One patient had short period of intraoperative oxygen desaturation. The mean operating time was 6.2 minutes with a range of 3 to 10 minutes. Eight procedures were performed without any difficulty, 2 procedures were performed with some difficulties which could be managed by the surgeon. Six patients had been examined under fibrobronchoscopy within one week postoperatively, no posterior tracheal wall injuries were found. One patient had peristomal bleeding after the operation, one patient had mild infection of the tracheostoma. There were no life threatening complications attributable to this technique. CONCLUSIONS: Controlled rotating dilation is a simple, rapid and safe bedside procedure. In the absence of bronchoscopic guidance, the procedure can be safely performed with precautions.