[Effects of urogenital mycoplasma and chlamydial infections on male fertility].

Objective： This study aimed to assess the impact of urogenital ureaplasma urealyticum (Uu), Mycoplasma hominis (Mh), and Chlamydia trachomatis (Ct) infections on semen quality in men.Methods： In this study, 1022 males were enrolled at the Department of Reproductive Medicine, Rizhao People's Hospital, Shandong Province from October 2014 to January 2023. The participants included 393 in the infertility group, 139 in the recurrent miscarriage group, and 490 in the control group. Based on age, 852 cases were < 36 years old, and 170 cases were ≥ 36 years old. All patients underwent routine semen analysis and tests for Uu, Mh, and Ct, with results statistically analyzed for their impact on semen quality and compared among different age groups. Results: Among the 1022 patients, 344 (33.6%) were Uu-positive, 49 (4.7%) were Mh-positive, and 31 (3.0%) were Ct positive. The sperm concentration, total sperm count, forward sperm motility rate (PR), sperm motility rate (PR+NP) and normal sperm morphology rate of Uu Mh and/or Ct-positive patients were significantly lower than those of the negative group, and the overall difference between the two groups was statistically significant (P<0.05). The positive rate of Uu infection was 41.7% in the infertility group, 30.2% in the recurrent miscarriage group and 28.2% in the control group, and the overall positive rate of the three groups was statistically significant(P<0.05). The positive rate of Mh infection was 6.9% in the infertility group, 8.6% in the recurrent miscarriage group and 2.0% in the control group, and the overall positive rate of the three groups was statistically significant (P<0.001). The positive rate of Ct infection was 6.1% in the infertility group, 2.9% in the recurrent miscarriage group and 0.6% in the control group, and the overall positive rate of the three groups was statistically significant (P<0.001). The positivity rate of Uu infection was 35.8% at the age of <36 years and 22.9% at the age ≥ 36 years, and there was a statistically significant difference in the positivity rate between the two groups (P<0.05). Conclusion: The prevalence of Uu infection in the male urogenital tract is significantly higher than that of Mh and Ct, which is the main pathogen of urogenital tract infection in men. Uu, Mh and Ct infections have adverse effects on sperm concentration, total sperm count, sperm forward motility rate (PR), sperm motility rate (PR+NP) and normal sperm morphology rate, which will lead to a decrease in semen quality and affect male fertility. Genital tract infections are closely related to age, and the prevalence of Uu infection is higher in the younger age group.

Efficient production of isoflavonoids by Astragalus membranaceus hairy root cultures and evaluation of antioxidant activities of extracts.

In this study, Astragalus membranaceus hairy root cultures (AMHRCs) were established as an attractive alternative source for the efficient production of isoflavonoids (IF). A. membranaceus hairy root line II was screened as the most efficient line and was confirmed by PCR amplification of rolB, rolC and aux1 genes. Culture parameters of AMHRCs were systematically optimized, and five main IF constituents were quali-quantitatively determined by LC-MS/MS. Under optimal conditions, the total IF accumulation of 34 day old AMHRCs was 234.77 µg/g dry weight (DW). This yield was significantly higher compared to that of 3 year old field grown roots (187.38 µg/g DW). Additionally, in vitro antioxidant assays demonstrated that AMHRC extracts exhibited antioxidant activities with lower IC50 values (1.40 and 1.73 mg/mL) as compared to those of field grown roots (1.96 and 2.17 mg/mL). Overall, AMHRCs may offer a promising and continuous product platform for naturally derived, high quality and valuable nutraceuticals.

PA-1, a novel synthesized pyrrolizidine alkaloid, inhibits the growth of Escherichia coli and Staphylococcus aureus by damaging the cell membrane.

In the present study, antimicrobial activity and mode of a novel synthesized pyrrolizidine alkaloid (PA-1) were investigated. PA-1 exhibited predominantly strong antibacterial activity toward six bacteria tested with minimal inhibitory concentration (MIC) values ranging from 0.0039 to 0.025 mg ml(-1). The time-kill assay indicated that PA-1 killed Escherichia coli and Staphylococcus aureus completely at 2MIC (minimum bactericidal concentration) within 8 h. Besides, PA-1-induced death rates of most sensitive strains (E. coli, 97.80% and S. aureus, 96.24%) were analyzed by flow cytometry. A combination of approaches was used to verify the membrane damage of E. coli and S. aureus. Results showed that release of 260 nm absorbing materials quickly increased after PA-1 treatment. PA-1 also rapidly promoted the uptake of crystal violet from 24.52 to 97.12% for E. coli and from 19.68 to 97.63% for S. aureus when the concentrations were changed from MIC to 4MIC. Furthermore, the cellular membrane damages were testified by the significant increase of fluorescence intensity and decrease of membrane potential. Finally, lecithin and phosphate groups were applied to search the possibly targets on the cytoplasmic membrane. Results showed that PA-1 acted on cytoplasmic membrane phospholipids and phosphate groups of S. aureus but not of E. coli. In conclusion, the novel synthesized PA-1 exerted its antibacterial activity by acting on membrane phospholipids and phosphate groups and then damaging the structures of cellular membrane, which finally led to cell death.