To determine the role of outer dense fiber (ODF) in multiple morphological abnormalities of the sperm flagella in Akap4 gene defect mice.
 Methods: Akap4 knock-out (KO) mouse model was established by using gene editing technology. Akap4-KO male mice were identified by genotype. Seven sexually mature male Akap4-KO mice served as an experimental group, and 7 sexually mature wild-type (WT) male mice served as a control group. The changes in body weight and testicular weight were measured. Computer aided sperm analysis (CASA) was used to detect sperm motility. Sperm morphology was detected by modified Periodic Acid-Schif (PAS) staining. The ultra-structure of sperm was observed under the scanning and transmission electron microscope. Sperm flagella associated protein expression and localization were detected by immunofluorescence. Spermatogenesis function of testis was evaluated by HE and PAS staining. Ultra-structure of seminiferous tubules was observed under the transmission electron microscope.
 Results: Akap4-KO mice had no natural fertility. The sperm motility of Akap4-KO male mice was lower than that of WT male mice (8.81% vs 46.02%, P<0.01). In Akap4-KO male mice the percentage of sperm, with shortened tail and coiled tail was 91.18% which was higher than that of WT male mice (P<0.01). There was no statistically significance in the testicular weight, spermatogensis function, and sperm count between the 2 groups (P>0.01). The longitudinal column of fibrous sheath in Akap4-KO male mice was absent, and the residues of transverse rib remained, which was consistent with the immunofluorescence localization of AKAP3 protein. No. 3 and No. 8 ODF in the principal piece were disordered, which was in consistent with ectopic localization of ODF2 protein.
 Conclusion: Multiple morphological abnormalities of the sperm flagella in mice are resulted from disorder of "9+2" microtubules and the abnormally expanded lumen at the proximal of the principal piece via causing dysplasia of the transverse rib due to Akap4 gene defect, and separation of the ODF of No. 3 and No. 8 via loss of longitudinal column.
A Kinase Anchor Proteins ; Animals ; Fluorescent Antibody Technique ; Infertility, Male ; Male ; Mice ; Sperm Motility ; Sperm Tail ; Spermatozoa

Background: High-dose dual therapy (HDDT) is a novel regimen for the eradication of Helicobacter pylori (Hp) infection, however, its efficacy and safety remain unclear. Aims: To evaluate the efficacy, safety and compliance of HDDT for Hp eradication. Methods: Randomized controlled trials (RCTs) on HDDT for eradication of Hp infection were retrieved from PubMed, Embase, The Cochrane Library, Web of Science from the date of database establishment to October 2020. Literatures were enrolled according to the inclusion and exclusion criteria, and the data were extracted. RevMan 5.2 software was used for performing meta-analysis. Results: Nine RCTs including 2 627 patients were included. Meta-analysis results showed that no significant differences in ITT eradication (85.4% vs. 79.8%, RR=1.03, 95% CI: 0.96-1.10, P=0.40), PP eradication (88.7% vs. 83.4%, RR=1.01, 95% CI: 0.95-1.08, P=0.68), and compliance (96.5% vs. 95.9%, RR=1.01, 95% CI: 0.99-1.02, P=0.37) were found between HDDT and the guideline-recommended regimens, however, the incidence of adverse events was significantly decreased in HDDT (15.3% vs. 27.0%, RR=0.57, 95% CI: 0.42-0.76, P=0.000 2). Conclusions: There are no significant differences in eradication rates and compliance between HDDT and the guideline-recommended regimens, however, HDDT is much safer.

Danshen, the dried roots and rhizomes of Salvia miltiorrhiza Bunge (S. miltiorrhiza), is widely used in the treatment of cardiovascular and cerebrovascular diseases. Tanshinones, the bioactive compounds from Danshen, exhibit a wide spectrum of pharmacological properties, suggesting their potential for future therapeutic applications. Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized, most of which belong to the CYP76 and CYP71 families. In this study, CYP81C16, a member of the CYP71 clan, was identified in S. miltiorrhiza. An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones, namely neocryptotanshinone, deoxyneocryptotanshinone, and danshenxinkuns A and B. SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%. Kinetic evaluations andin vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates. The overexpression of SmCYP81C16 promoted the accumulation of (iso)tanshinone in hairy root lines. The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones, either through microbial or plant metabolic engineering.
Humans ; Salvia miltiorrhiza/metabolism* ; Biosynthetic Pathways ; Quinones/metabolism* ; Plant Roots/metabolism* ; Gene Expression Regulation, Plant