Identification of compound heterozygous variants in MSH4 as a novel genetic cause of diminished ovarian reserve.

BACKGROUND: Diminished ovarian reserve (DOR) is a common cause of female infertility, with genetic factors being a significant contributor. However, due to high genetic heterogeneity, the etiology of DOR in many cases remains unknown. In this study, we analyzed the phenotype of a young woman with primary infertility and performed molecular genetic analysis to identify the genetic cause of her condition, thus providing important insights for genetic counseling and reproductive guidance. METHODS: We collected the patient's basic information, clinical data, as well as diagnostic and therapeutic history and performed whole-exome sequencing on her peripheral blood. Candidate pathogenic variants were validated by Sanger sequencing in family members, and the pathogenicity of variants was analyzed using ACMG guidelines. We used bioinformatics tools to predict variant effects on splicing and protein function, and performed in vitro experiments including minigene assay and expression analysis to evaluate their functional effects on HEK293T. RESULTS: We identified biallelic MSH4 variants, c.2374 A > G (p.Thr792Ala) and c.2222_2225delAAGA (p.Lys741Argfs*2) in the DOR patient. According to ACMG guidelines, the former was classified as likely pathogenic, while the latter was classified as pathogenic. The patient presented with poor oocyte quantity and quality, resulting in unsuccessful in vitro fertilization cycles. Bioinformatics and in vitro functional analysis showed that the c.2374 A > G variant altered the local conformation of the MutS_V domain without decreasing MSH4 protein expression, while the c.2222_2225delAAGA variant led to a reduction in MSH4 protein expression without impacting splicing. CONCLUSIONS: In this study, we present evidence of biallelic variants in MSH4 as a potential cause of DOR. Our findings indicate a correlation between MSH4 variants and reduced oocyte quality, as well as abnormal morphology of the first polar body, thereby expanding the phenotypic spectrum associated with MSH4 variants. Furthermore, Our study emphasizes the importance of utilizing whole-exome sequencing and functional analysis in diagnosing genetic causes, as well as providing effective genetic counseling and reproductive guidance for DOR patients.

Effect of peer videorecording feedback CPR training on students' practical CPR skills: a randomized controlled manikin study.

BACKGROUND: The aim of this study was to compare one-month acquisition and half-a-year quality retention of cardiopulmonary resuscitation (CPR) skills after initial training of medical students between peer videorecording feedback training (PVF) and traditional peer verbal feedback training (TVF). METHODS: Participants were randomly assigned to the PVF group (n = 62) and the TVF group (n = 65). All participants received a 45-min CPR training program performed by an American Heart Association basic life support-certified instructor, and then they began two hours of practice in groups. During interactive peer learning, students cooperated in couples of a doer and a helper to realize maximization of each other's learning. In the PVF group, training performance feedback came from peers based on practice videorecording. In the TVF group, feedback came from peers verbally without videorecording. CPR quality was tested at 1 and 6 months after training. RESULTS: After 1 month of initial training, the PVF group had a better presentation of CPR skills acquisition than the TVF group. Compared to the TVF group, the PVF group had significantly higher total scores, compression depth, appropriate compression depth, and complete chest recoil (p < 0.05). Moreover, compression interruption was a significantly positive change in the PVF group compared to the TVF group (p < 0.05). However, after 6 months, proportions of appropriate compression depth in the PVF group were better than those in the TVF group (p < 0.05). The differences in total scores, compression depth, appropriate compression depth, complete chest recoil and compression interruption were non-significant (all p > 0.05). CONCLUSIONS: Compared to TVF, PVF is more effective in enhancing CPR skill acquisition at 1 month. After half a year, CPR skill quality was obviously reduced in both groups, and no difference in CPR quality was found between the two groups.

Effect of Different Influent Conditions on Biomass Production and Nutrient Removal by Aeration Microalgae Membrane Bioreactor (ICFB-MMBR) System for Mariculture Wastewater Treatment.

The nutrient removal and biomass production of the internal circulating fluidized bed microalgae membrane bioreactor (ICFB-MMBR) was studied under different cultivation modes, influent TOC, influent pH, and influent N/P. Platymonas helgolandica tsingtaoensis was used as the biological source. The growth of P. helgolandica tsingtaoensis and the removal efficiency of pollutants in the mixotrophy culture mode were improved compared with other culture modes. With the increased influent TOC, the average growth rate of P. helgolandica tsingtaoensis increased, and ammonia nitrogen and total phosphorus removal rate were improved. The P. helgolandica tsingtaoensis growth rate and nutrient removal efficiencies at the influent pH of 8 were the best among the different influent pH values. As the influent N/P ratio increased from 5 to 20, the P. helgolandica tsingtaoensis growth rate and pollutant removal rate increased gradually. When the influent N/P ratio was higher than 20, the P. helgolandica tsingtaoensis growth rate and pollutant removal rate tended to be stable and did not significantly change with the increase of influent N/P ratio. At the proper influent conditions, the high P. helgolandica tsingtaoensis biomass and nutrient removal efficiency could be obtained in the microalgae membrane bioreactor, which could provide a theoretical basis for the application of the system for wastewater treatment.

The Influence of Different Operation Conditions on the Treatment of Mariculture Wastewater by the Combined System of Anoxic Filter and Membrane Bioreactor.

The mariculture wastewater treatment performance for the combined system of anoxic filter and membrane bioreactor (AF-MBR) was investigated under different hydraulic retention times (HRTs), influent alkalinity, and influent ammonia nitrogen load. The results showed that the removal efficiencies of TOC and total nitrogen were slightly better at the HRT of 8 h than at other HRTs, and the phosphate removal efficiency decreased with the increase of HRT. With the increase of influent alkalinity, the removal of TOC and phosphate did not change significantly. With the increase of influent alkalinity from 300 mg/L to 500 mg/L, the total nitrogen removal efficiency of AF-MBR was improved, but the change of the removal efficiency was not obvious when the alkalinity increased from 500 mg/L to 600 mg/L. When the influent concentration of ammonia nitrogen varied from 20 mg/L to 50 mg/L, the removal efficiencies of TOC, phosphate, and total nitrogen by AF-MBR were stable. An interesting finding was that in all the different operation conditions examined, the treatment efficiency of AF-MBR was always better than that of the control MBR. The concentrations of NO3--N in AF-MBR were relatively low, whereas NO3--N accumulated in the control MBR. The reason was that the microorganisms attached to the carrier and remained fixed in the aerobic and anoxic spaces, so that there was a gradual enrichment of bacteria characterized by slow growth in a high-salt environment. In addition, the microorganisms could gather and grow on the carrier forming a biofilm with higher activity, a richer and more stable population, and enhanced ability to resist a load impact.

Impacts of bio-carriers on the characteristics of soluble microbial products in a hybrid membrane bioreactor for treating mariculture wastewater.

To gain greater insights into impacts of bio-carriers on the fate and characteristics of soluble microbial products (SMPs) for mariculture wastewater treatment, the hybrid membrane bioreactor (HMBR) and conventional membrane bioreactor (CMBR) were investigated. Both protein and polysaccharide exhibited lower level in HMBR (8.95 ± 0.28 mg/L and 20.49 ± 1.3 mg/L for anoxic stage, 5.16 ± 0.22 mg/L and 17.85 ± 0.92 mg/L for aerobic stage) than CMBR (14.6 ± 0.68 mg/L and 28.3 ± 2.99 mg/L for anoxic stage, 10.53 ± 0.68 and 26.04 ± 3.15 mg/L for aerobic stage). Three-dimensional fluorescence excitation emission matrix (EEM) revealed bio-carriers reduced the production of aromatic protein-like components in anoxic and aerobic supernatant and caused a blue-shift of soluble microbial product in aerobic stage. Molecular weight (Mw) distribution indicated that bio-carriers ameliorated the excretion of biopolymer (Mw > 500 kDa) in anoxic supernatant and intermediate Mw fractions (20-500 kDa) in aerobic supernatant. Moreover, little changes were observed in SMPs with Mw < 3 kDa down the whole treatment process of both systems. Gas chromatography-mass spectrometry (GC-MS) demonstrated that the major SMPs were long-chain alkanes and aromatics in all units of both systems and fewer aromatics were detected in HMBR. For anoxic stage, more peaks were identified in the HMBR (138) than CMBR (115), while for aerobic stage, more compounds were observed in the CMBR (94) than HMBR (70). Over 50% of the compounds in the anoxic supernatant for the HMBR were the same as in the CMBR. And 27 compounds were the same in aerobic supernatant for the HMBR and CMBR. Fewer compounds in the HMBR effluent (52) was observed, compared to CMBR effluent (80). Approximately 25.7% of compounds in the aerobic stage of the HMBR were rejected by membrane, while this value decreased to 14.9% in the CMBR.