Rhythm gene PER1 mediates ferroptosis and lipid metabolism through SREBF2/ALOX15 axis in polycystic ovary syndrome.

OBJECTIVE: This work aimed to investigate the role of rhythm gene PER1 in mediating granulosa cell ferroptosis and lipid metabolism of polycystic ovary syndrome (PCOS). METHODS: We injected dehydroepiandrosterone and Ferrostatin-1 (Fer-1) into mice to explore the mechanism of ferroptosis in PCOS. The effect of PER1 on ferroptosis-like changes in granulosa cells was explored by overexpression of PER1 plasmid transfection and Fer-1 treatment. RESULTS: We found that Fer-1 ameliorated the characteristic polycystic ovary morphology, suppressed ferroptosis in the PCOS mice. PER1 and ALOX15 were highly expressed in PCOS, whereas SREBF2 was lowly expressed. Overexpression of PER1 decreased granulosa cell viability and inhibited proliferation. Meanwhile, overexpression of PER1 increased lipid reactive oxygen species, 4-Hydroxynonenal (4-HNE), Malondialdehyde (MDA), total Fe, and Fe2+ levels in granulosa cells and decreased Glutathione (GSH) content. Fer-1, SREBF2 overexpression, or ALOX15 silencing treatment reversed the effects of PER1 overexpression on granulosa cells. PER1 binds to the SREBF2 promoter and represses SREBF2 transcription. SREBF2 binds to the ALOX15 promoter and represses ALOX15 transcription. Correlation analysis of clinical trials showed that PER1 was positively correlated with total cholesterol, low-density lipoprotein cholesterol, luteinizing hormone, testosterone, 4-HNE, MDA, total Fe, Fe2+, and ALOX15. In contrast, PER1 was negatively correlated with SREBF2, high-density lipoprotein cholesterol, follicle-stimulating hormone, progesterone, and GSH. CONCLUSION: This study demonstrates that the rhythm gene PER1 promotes ferroptosis and dysfunctional lipid metabolism in granulosa cells in PCOS by inhibiting SREBF2/ALOX15 signaling.

E. coli Nissle 1917 ameliorates mitochondrial injury of granulosa cells in polycystic ovary syndrome through promoting gut immune factor IL-22 via gut microbiota and microbial metabolism.

Objective: Gut microbiota and its metabolites have regulatory effects on PCOS related ovarian dysfunction and insulin resistance. Escherichia coli Nissle 1917 (EcN) is a genetically controlled probiotic with an excellent human safety record for improving gut microbiome metabolic disorders and immune system disorders. Here we focused to explore the application and effect of probiotic EcN on the gut microbiota-metabolism-IL-22-mitochondrial damage axis in PCOS. Methods: PCOS mice were constructed with dehydroepiandrosterone (DHEA) and treated with EcN, FMT or IL-22 inhibitors. Clinically control and PCOS subjects were included for further analysis. Serum and follicular fluid supernatant levels of sex hormones, insulin, glucose, cholesterol, and inflammatory factors were detected by ELISA and biochemical reagents. The pathological changes of ovarian tissues were observed by HE staining. The JC-1 level and COX4 gene expression in granulosa cells was detected by ELISA and RT-qPCR. The expressions of progesterone receptor A (PR-A), LC3II/I, Beclin1, p62 and CytC were detected by western blot. The number of autophagosomes in granulosa cells was observed by electron microscopy. 16S rRNA and LC-MS/MS were used to analyze the changes of gut microbiota and metabolism. Results: EcN promoted the recovery of sex hormone levels and ovarian tissue morphology, promoted the expression of IL-22, COX4 and PR-A in granulosa cells, and inhibited mitophagy in PCOS mice. EcN decreased the number of gut microbiota, and significantly increased the abundance of Adlercreutzia, Allobaculum, Escherichia-Shigella and Ileibacterium in PCOS mice. EcN improved metabolic disorders in PCOS mice by improving Amino sugar and nucleotide sugar metabolism pathways. IL-22 was positively associated with Ileibacterium, Adlercreutzia and Progesterone, negatively associated with RF39, Luteinizing hormone, Testosterone, N-Acetylglucosamin, L-Fucose and N-Acetylmannosamin. FMT reconfirmed that EcN ameliorated mitochondrial damage in granulosa cells of PCOS mice by gut microbiota, but this process was blocked by IL-22 inhibitor. Clinical trials have further demonstrated reduced IL-22 levels and mitochondrial damage in granulosa cells in PCOS patients. Conclusion: EcN improved IL-22 level and mitochondrial damage of granulosa cells in PCOS mice by promoting the recovery of sex hormone levels and ovarian tissue morphology, inhibiting the amount of gut microbiota, and promoting amino sugar and nucleotide sugar metabolism.

[Correlation of sperm HDS with sperm DFI and morphology and its influence on in vitro fertilization in infertile males].

OBJECTIVE: To invest the correlation of sperm high DNA stainability (HDS) with sperm DNA fragmentation index (DFI) and sperm abnormalities and its influence on in vitro fertilization (IVF) in male infertility patients, and assess the clinical value of HDS. METHODS: Using flow cytometry-assisted sperm chromatin structure assay (SCSA), we examined sperm HDS and sperm DFI in 322 male infertility patients undergoing IVF due to female fallopian tube factors only. Based on sperm HDS, we divided the patients into five groups and compared the semen routine parameters, percentage of morphologically abnormal sperm (MAS), sperm DFI, rates of fertilization, cleavage and high-quality embryos, and pregnancy outcomes among different HDS groups. RESULTS: Among the 322 male infertility patients, 119 (36.96%) were found with a sperm HDS of 0 － <5%, 117 (36.34%) of 5% － <10%, 50 (15.53%) of 10% － <15%, 23 (7.14%) of 15% － <20%, and 13 (4.03%) of ≥20%. Sperm concentration, motility and progressive motility were decreased with the increase of sperm HDS, but with no statistically significant difference (P > 0.05), so were the rates of fertilization, high-quality embryos and pregnancy (P > 0.05). Sperm DFI and sperm abnormality were correlated positively with sperm HDS (r = 0.236, r = 0.203). The rate of early abortion was remarkably increased in those with sperm HDS greater than 10%. CONCLUSION: Sperm HDS may be a risk indicator of sperm DFI and sperm abnormality, and can be used as a predictive indicator of early abortion in IVF.

[Correlation of sperm DNA fragmentation index and high DNA stainability with teratospermia severity and the patient's age: A study of 1 393 infertile males].

OBJECTIVE: To investigate the correlation of the severity of teratospermia and the age of the patient with sperm DNA fragmentation index (DFI) and high DNA stainability (HDS) in male infertility patients. METHODS: We collected semen samples from 1 393 infertile males from July to December 2021. Based on the WHO Laboratory Manual for the Examination and Processing of Human Semen (5th Ed), we performed sperm morphology analysis, examined perm DFI and HDS by flow cytometry, and analyzed the impacts of the severity of teratospermia and the age of the patients on sperm DFI and HDS. RESULTS: Among the 1 393 male infertility patients, 124 (8.90%) were found with extremely severe, 214 (15.36%) with severe, 235 (16.87%) with moderate, 163 (11.70%) with mild teratospermia, and 657 (47.16%) with morphologically normal sperm (MNS), with statistically significant differences in sperm DFI and HDS among the five groups, and 822 (59.00%) were aged <35 years, 306 (21.97%) 35－<40 years, 223 (16.01%) 40－<45 years and 42 (3.02%) ≥45 years, with statistically significant differences in sperm DFI and HDS among different age groups (P < 0.05). Sperm DFI and HDS were correlated negatively with the percentage of MNS (P > 0.05), but positively with the age of the patients (P < 0.05). CONCLUSION: Increased severity of teratospermia and age of the patient can increase sperm DFI and HDS, and sperm nuclear chromatin integrity and maturity are important indicators of male fertility.

Mechanism of Chinese Medicine in Induction of Apoptosis of Lung Cancer Cells： A Review / 中国实验方剂学杂志

Lung cancer， a malignancy with high incidence rate and mortality rate， is a major threat to human life and health. At present， the common methods for the treatment of lung cancer include surgical resection， radiotherapy， chemotherapy， targeted therapy， and immunotherapy， but these methods generally have the problems of severe toxic/side effect and high treatment cost. Traditional Chinese medicine（TCM） has a history of more than 2 000 years of application in China and has its unique advantages in the treatment of tumors. Modern pharmacological experiments have found that TCM can inhibit tumor growth， prolong patients' survival， and improve clinical symptoms and patients' quality of life by inducing tumor cell apoptosis， inhibiting tumor angiogenesis， and reducing tumor cell drug resistance. Apoptosis is a process of spontaneous programmed cell death， which is closely related to the occurrence and development of the tumor. Studies have shown that many Chinese medicines can inhibit the development of lung cancer by inducing apoptosis. This study searched， analyzed， and summarized the available papers on the mechanism of TCM in the treatment of lung cancer by inducing apoptosis. It is found that Chinese medicine induces lung cancer cell apoptosis mainly by regulating apoptosis-related factors and apoptosis-related signaling pathways ［inhibitor of apoptosis proteins （IAPs）， B cell lymphoma-2 （Bcl-2）， p53 protein， the second mitochondria-derived activator of caspase （SMAC）/direct IAP-binding protein with low isoelectric point （DIABLO）， extrinsic apoptotic pathway， endogenous mitochondrial pathway， Janus kinase （JAK）/signal transducer and activator of transcription （STAT） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， and phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. In addition， the Wnt/β-catenin/survivin signaling pathway and the Notch signaling pathway also play an important role in inducing apoptosis.

Updates and advancements in the management of hepatocellular carcinoma patients after hepatectomy.

Introduction: The 5-year recurrence rate of hepatocellular carcinoma (HCC) after hepatic resection or local ablation is up to 70%. Adjuvant therapies to prevent HCC recurrence have been reported but are not currently recommended by EASL or AASLD guidelines. This review examined evidence from randomized controlled trials, meta-analyses and systematic reviews on the safety and efficacy of adjuvant therapies and chemotherapies in HCC patients after resection or local ablation.Areas covered: PubMed was searched through 15 June 2019. Available evidence was assessed based on the GRADE system.Expert commentary: Transarterial chemoembolization is the best adjuvant therapy for HCC patients at high risk of recurrence, antiviral therapy with nucleoside analogs is effective for preventing recurrence of HBV-related HCC, and interferon-α is effective for preventing recurrence of HCV-related HCC. Further studies are needed to clarify the efficacy of adjuvant immune checkpoint inhibitors. Adjuvant sorafenib appears to offer negligible clinical benefit and high risk of adverse effects.

[Optimization of sperm alkaline single-cell gel electrophoresis].

OBJECTIVE: To investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures. METHODS: Using alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method. RESULTS: Three agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05). CONCLUSION: The optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.

Multifunctional homopolymer vesicles for facile immobilization of gold nanoparticles and effective water remediation.

Homopolymers have been considered as a nonideal building block for creating well-defined nanostructures due to their fuzzy boundary between hydrophobic and hydrophilic moieties. However, this unique fuzzy boundary may provide some opportunities for fabricating functional nanomaterials. Presented in this paper is a pH-responsive multifunctional homopolymer vesicle based on poly[2-hydroxy-3-(naphthalen-1-ylamino)propyl methacrylate] (PHNA). This vesicle is confirmed to be an excellent supporter for gold nanoparticles (AuNPs) to facilitate the reduction reaction of 4-nitrophenol (4-NP). The pH-responsive vesicle membrane favors the effective embedding and full immobilization of AuNPs because it is kinetically frozen under neutral and basic environments, preventing AuNPs from aggregation. Meanwhile, there is a synergistic effect between the AuNPs and the supporter (PHNA vesicle). Due to the π-π interaction between the naphthalene pendants in every repeat unit of PHNA and the extra aromatic compounds, a substrate-rich (high concentration of 4-NP) microenvironment can be created around AuNPs, which can dramatically accelerate the AuNPs-catalyzed reactions. In addition, we proposed a method for more accurately determining the membrane thickness of rigid polymer vesicles from TEM images based on "stack-up" vesicles, which may overturn the measuring method commonly used by far. Moreover, proof-of-concept studies showed that those homopolymer vesicles may be used as a powerful adsorbent for effective water remediation to remove trace carcinogenic organic pollutants such as polycyclic aromatic hydrocarbons to below parts per billion (ppb) level at a very fast rate based on the π-π interaction between the naphthalene pendants in PHNA vesicle and polycyclic aromatic hydrocarbons. Overall, this multifunctional homopolymer vesicle provides an alternative insight on preparing effective recyclable AuNPs-decorated nanoreactor and powerful water remediation adsorbent.

Homopolymer vesicles with a gradient bilayer membrane as drug carriers.

We report an unusual homopolymer vesicle, which has a soft bilayer membrane composed of oligo(ethyleneoxy) side chains (OEs) with a gradually decreased packing density from the centre of the membrane to both margins, exhibiting thermo-responsive zeta potential and dispersibility and showing potential applications for anti-cancer drug delivery.

Water-dispersible and biodegradable polymer micelles with good antibacterial efficacy.

New amphiphilic ABC triblock copolymers have been designed and self-assembled into water-dispersible and biodegradable polymer micelles, which exhibit good antibacterial activities without quaternary ammonium moieties or the loading of any external antibiotics due to the increased local concentration of cationic charge in the polymer micelles compared to the un-self-assembled individual polymer chains.

Pulmonary valve endocarditis in a pregnant woman with a ventricular septal defect.

Bacterial endocarditis in pregnancy causes maternal and fetal mortality rates of 22.1% and 14.7%, respectively. The mortality rates differ according to the involved valves, and the size of vegetation has a prognostic correlation. This report is of a pregnant woman with an unrepaired ventricular septal defect and pulmonary valve endocarditis with a vegetation size of 3.29 cm. She and her baby were treated successfully. An emergency surgical plan would be appropriate for pregnant women in the third trimester with a large vegetation in the right side of the heart, and dental disease should be treated aggressively with appropriate prophylactic antibiotics.