Significant decrease of maternal mitochondria carryover using optimized spindle-chromosomal complex transfer.

Mutations in mitochondrial DNA (mtDNA) contribute to a variety of serious multi-organ human diseases, which are strictly inherited from the maternal germline. However, there is currently no curative treatment. Attention has been focused on preventing the transmission of mitochondrial diseases through mitochondrial replacement (MR) therapy, but levels of mutant mtDNA can often unexpectedly undergo significant changes known as mitochondrial genetic drift. Here, we proposed a novel strategy to perform spindle-chromosomal complex transfer (SCCT) with maximal residue removal (MRR) in metaphase II (MII) oocytes, thus hopefully eliminated the transmission of mtDNA diseases. With the MRR procedure, we initially investigated the proportions of mtDNA copy numbers in isolated karyoplasts to those of individual oocytes. Spindle-chromosomal morphology and copy number variation (CNV) analysis also confirmed the safety of this method. Then, we reconstructed oocytes by MRR-SCCT, which well developed to blastocysts with minimal mtDNA residue and normal chromosomal copy numbers. Meanwhile, we optimized the manipulation order between intracytoplasmic sperm injection (ICSI) and SCC transfer and concluded that ICSI-then-transfer was conducive to avoid premature activation of reconstructed oocytes in favor of normal fertilization. Offspring of mice generated by embryos transplantation in vivo and embryonic stem cells derivation further presented evidences for competitive development competence and stable mtDNA carryover without genetic drift. Importantly, we also successfully accomplished SCCT in human MII oocytes resulting in tiny mtDNA residue and excellent embryo development through MRR manipulation. Taken together, our preclinical mouse and human models of the MRR-SCCT strategy not only demonstrated efficient residue removal but also high compatibility with normal embryo development, thus could potentially be served as a feasible clinical treatment to prevent the transmission of inherited mtDNA diseases.

Phytochemical analysis and hepatotoxicity assessment of braised Polygoni Multiflori Radix (Wen-He-Shou-Wu).

Polygoni Multiflori Radix (PMR) is a medicinal herb commonly used in China and Eastern Asia. Recently, the discovery of hepatotoxicity in PMR has received considerable attention from scientists. Processing is a traditional Chinese medicine technique used for the effective reduction of toxicity. One uncommon technique is the braising method-also known as 'Wen-Fa' in Chinese-which is used to prepare tonics or poisonous medications. Braised PMR (BPMR)-also known as 'Wen-He-Shou-Wu'-is one of the processed products of the braising method. However, the non-volatile components of BPMR have not been identified and examined in detail, and therefore, the hepatotoxic advantage of BPMR remains unknown. In this study, we compared the microscopic characteristics of different samples in powder form using scanning electron microscopy (SEM), investigated the non-volatile components, assessed the effects of different processed PMR products on the liver, and compared the differences between BPMR and PMR Praeparata recorded in the Chinese Pharmacopoeia (2020 edition). We found that the hepatotoxicity of BPMR was dramatically decreased, which may be related to an increase in polysaccharide content and a decrease in toxic substances. The present study provides an important foundation for future investigations of the processing mechanisms of BPMR.

The volatile organic compounds generated from the Maillard reaction between l-ascorbic acid and l-cysteine in hot compressed water.

BACKGROUND: Hot compressed water (HCW), also known as subcritical water (SCW), refers to high-temperature compressed water in a special physical and chemical state. It is an emerging technology for natural product extraction. The volatile organic compounds (VOCs) generated from the Maillard reaction between l-ascorbic acid (ASA) and l-cysteine (Cys) have attracted significant interest in the flavor and fragrance industry. This study aimed to explore the formation mechanism of VOCs from ASA and Cys and examine the effects of reaction parameters such as temperature, time, and pH in HCW. RESULTS: The identified VOCs were predominantly thiophene derivatives, polysulfides, and pyrazine derivatives in HCW. The findings indicated that thiophene derivatives were formed under various pH conditions, with polysulfide formation favored under acidic conditions and pyrazine derivative formation preferred under weak alkaline conditions, specifically at pH 8.0. CONCLUSION: The Maillard reaction between ASA and Cys mainly produced thiophene derivatives, polysulfides, and pyrazine derivatives in HCW. The generation mechanism was significantly dependent on the surrounding pH conditions. © 2024 Society of Chemical Industry.

Living arrangements, activity participation and depression among older Chinese adults.

OBJECTIVES: We examined the relationship among living arrangements, activity participation and depression and the mediating effects of activity participation on the relationship between living arrangements and depression. STUDY DESIGN: This was a cross-sectional study based on nationally representative data. METHODS: 9647 older adults aged 60 years and over were recruited from the 2018 Chinese Longitudinal Healthy Longevity Survey. Three-step regression and mediation analysis methods were used. RESULTS: Compared with older adults living with family members, those living in institutions attended fewer activities and experienced higher depression rates, and those living alone attended more activities and experienced higher depression rates. In the group living with family members, living with a spouse was an important way to increase activity participation and reduce depression. Mediation analysis suggested that activity participation mediated the impact of living arrangements on depression. Attending more activities could alleviate depression for older adults who are living alone, living in institutions and living without a spouse. CONCLUSION: Our findings suggest an emphasis on the role of spouses and activity participation during depression interventions among older adults.

The health service use of aged rural-to-urban migrant workers in different types of cities in China.

BACKGROUND: The association between different types of cities and the use of health services by aged migrant workers in China has not been widely reported in previous studies. This article aims to focus on rural-to-urban migrant workers in China aged 50 years and older to examine the relationship between the region of these migrant workers' destination city (eastern, central or western) and migration city type (first-tier, second-tier, third-tier and smaller cities) and their use of health services (e.g., establishing health records, participating in health education, and seeking medical treatment when ill). METHODS: This study's data were obtained from China Migrants Dynamic Survey in 2017. A total of 14,732 rural-to-urban migrant workers aged 50 years and older were included in the analysis; 6,938 of the migrant workers were either ill or had recently experienced physical discomfort. A chi-square test and binary logistic regression were performed to explore the associations between these rural-urban migrants' destination cities and their use of health services. RESULTS: This study found that aged rural migrant workers who moved to the east or to first- or second-tier cities were less likely to establish health records, participate in health education programme, and seek medical care. CONCLUSIONS: Migrant destination cities are linked to the use of local health services by migrant workers aged 50 years and older in China. We found that aged migrant workers who migrated to relatively developed regions and cities accessed fewer health services. Such results signify that more attention should be paid to aged migrant workers' use of health services in economically developed regions and cities, to eliminate regional differences in healthcare inequality.

Study on the mechanism of treating COVID-19 with Shenqi Wan based on network pharmacology.

PURPOSE: Through the method of network pharmacology, the active components and targets of Shenqi Wan (SQW) were excavated, the relationship with novel Coronavirus pneumonia (COVID-19) was discussed, and the possible mechanism of SQW in the treatment of COVID-19 was revealed from the aspects of multicomponents, multitargets, and multipathways. METHODS: Firstly, the active components of SQW were screened from traditional Chinese medicine systems pharmacology database and analysis platform and the 2020 edition of Chinese Pharmacopeia, and the related targets of the components were obtained. Then the disease targets related to COVID-19 were screened from GeneCards and Online Mendelian Inheritance in Man. Venny was used to map the relationship between component-target and disease-target, and String was used to analyze the interaction of common targets. The network was constructed and analyzed by Cytoscape, the function of Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) genes was enriched by Metascape, and the molecular docking was verified by CB-Dock. RESULTS: Finally, 45 active components of SQW were obtained, and 72 potential targets were related to COVID-19, angiotensin-converting enzyme 2 (ACE2), interleukin (IL)-6, nitric oxide synthase (NOS3) and, C-reactive protein (CRP),may be the key targets. GO enrichment of 1715 projects, such as lipopolysaccharide stress response, active oxygen metabolism, positive regulation of cell migration, and other GO enrichment. About 136 KEGG pathways, tumor necrosis factor signaling pathway, IL-17 signaling pathway, hypoxia-inducible factor 1-α signaling pathway were obtained. Molecular docking showed that kaempferol, quercetin, luteolin, astragaloside, calyx isoflavone glucoside, matrine, and other COVID-19-related targets such as ACE2, chymotrypsin-like protease (3CLpro), papain-like protease (PLpro), prostaglandin-endoperoxide synthase 2 (PTGS2) have good binding ability. CONCLUSION: According to the above results, it is suggested that SQW may play a role in the treatment of COVID-19 by directly or indirectly combining kaempferol, quercetin, and luteolin with ACE2, 3CLpro, PLpro, and PTGS2 to regulate multiple biological functions and signaling pathways.

[Study on mechanism of combination of Platycodonis Radix and Lilii Bulbus with homology of medicine and food in treating pneumonia].

To investigate the potential molecular mechanism of the combination of Platycodonis Radix and Lilii Bulbus with the homology of medicine and food in the treatment of pneumonia by means of network pharmacology and in vitro verification experiment. Under the condition of bioavailability(OB)≥30% and drug-like(DL)≥0.18, the active components of Platycodonis Radix and Lilii Bulbus were screened in TCMSP database; the prediction targets of active components were searched from TCMSP, DrugBank and other databases, and the potential targets of pneumonia were obtained through GeneCards and OMIM database. The common targets were obtained by the intersection of drug and disease targets. The PPI network of common targets was constructed by STRING 11.0, and the core targets were obtained by topological analysis. Then the core targets received GO and KEGG analysis with use of WebGestalt and Metascape. The "component-target-pathway" network was constructed with the help of Cytoscape 3.7.1 software, and the component-target molecular docking verification was carried out with Discovery Studio 2016 software. Finally, the core targets and pathways were preliminarily verified in vitro. In this study, 12 active components were screened, 225 drug prediction targets and 420 potential diseases targets were obtained based on data mining method, and 14 core targets were obtained by topological analysis, including TNF, MMP9, AKT1, IL4 and IL2. The enrichment results of GO and KEGG showed that "Platycodonis Radix and Lilii Bulbus" drug pair may regulate inflammation, cell growth and metabolism by acting on 20 key signaling pathways such as TNF and IL-17, thereby exerting anti-pneumonia effects. The results of molecular docking showed that 12 active components had good binding ability with 14 core targets. In vitro experiment results showed that the core components of "Platycodonis Radix and Lilii Bulbus" drug pair could inhibit the expression of MMP9 and TNF-α by regulating TNF signal pathway. This study confirmed the scientificity and reliability of the prediction results of network pharmacology, and preliminarily revealed the potential molecular mechanism of the compatibility of Platycodonis Radix and Lilii Bulbus in the treatment of pneumonia. It provides a novel insight on systematically exploring the mechanism of the compatible use of Platycodonis Radix and Lilii Bulbus, and has a certain reference value for the research, development and application of new drugs.

The ovulation trigger-OPU time interval of different ovarian protocols in ART: a retrospective study.

PURPOSE: To explore the trends of oocyte and pregnancy outcomes over the ovulation trigger-OPU (oocyte pickup) time interval in four mainly used COH protocols. METHODS: This retrospective study was conducted between January 2013 and July 2018. The IVF/ICSI cycles of the patients with normal ovarian reserve were included. The number of total patients was 4673, which consisted of long agonist protocol (n = 819), short agonist protocol (n = 1703), mild stimulation protocol (n = 1627), and GnRH antagonist protocol (n = 524). The primary outcome was mature oocyte rate. RESULTS: The ovulation trigger-OPU time interval and COH protocol were related to cycles with > 80% MII oocytes. Four protocols showed apparently different trends of retrieved oocyte rate and mature oocyte rate over the ovulation trigger-OPU time interval, and the long agonist protocol had the most delayed time interval than other three COH protocols in retrieving more than 60% oocytes (35.4-39.6 h vs. 34.6-38.6 h vs. 32.5-37.5 h vs. 33.8-37.7 h) and getting more than 80% mature oocytes (35.0-39.7 h vs. 36.0-37.7 h vs. 34.1-35.5 h vs. 34.5-36.3 h). And the adjusted odds ratio (OR) of the cumulative live birth rate (CLBR) (OR 1.360, 95% Confidence Interval (CI) 1.156-1.549, P < 0.05) significantly increased with the trigger-OPU time interval in the long agonist protocol. CONCLUSIONS: For getting more and mature oocytes, the ovulation trigger-OPU time intervals should be gradually prolonged from the mild stimulation protocol, the GnRH antagonist protocol, and the short protocol to the long agonist protocol. And the prolonged ovulation trigger-OPU time interval in the long agonist protocol brings higher live birth rate (LBR) and CLBR.

The cumulative live birth rates of 18 593 women with progestin-primed ovarian stimulation-related protocols and frozen-thawed transfer cycles.

STUDY QUESTION: What are the odds of achieving pregnancy when adopting progestin-primed ovarian stimulation (PPOS)-related protocols combined with repetitive frozen-thawed transfer (FET) cycles in patients with different clinical characteristics? SUMMARY ANSWER: The cumulative live birth rates (CLBRs) of women undergoing different PPOS-related protocols can be significantly and consistently enhanced within six FET cycles when the female age is <40 years (or even <45 years) and when >5 oocytes are retrieved, regardless of antral follicle count (AFC). WHAT IS KNOWN ALREADY: There have been numerous studies on the live birth rate of the first FET cycle in patients with PPOS-related protocols. These studies have focused mainly on comparing pregnancy outcomes with those of other stimulation protocols. However, owing to the unique features of the PPOS-related strategy, such as its flexible timing of oocyte retrieval and repeated transfer of frozen embryos, studies using the CLBR as an overall indicator of success and investigating which types of patients would benefit from this protocol are lacking. STUDY DESIGN SIZE DURATION: This retrospective cohort study included 18 593 women who underwent PPOS-related protocols (dydrogesterone + hMG, medroxyprogesterone acetate + hMG, micronized progesterone + hMG treatment, and luteal-phase ovarian stimulation protocol) from 1 March 2011 to 31 September 2022 in our centre. PARTICIPANTS/MATERIALS SETTING METHODS: The population was categorized by female age, number of oocytes retrieved, and AFC in the analysis of CLBR within six FET cycles. The age groups (Groups 1-5, respectively) were <30, 30-34, 35-39, 40-44, and ≥45 years. The number of oocytes retrieved was grouped as 1-5, 6-10, 11-15, 16-20, and >20. AFC was grouped as <5, 5-10, 11-15, and >15. The Kaplan-Meier analysis (optimistic method), which hypothesized that patients who did not continue treatment had the same chance of achieving a live birth as those who continued, and the competing risk method (conservative method) which hypothesized they had no chance of achieving a live birth, were applied. In further analyses, the Cox model and Fine-Gray model were adopted: the former corresponds to the optimistic scenario, and the latter corresponds to the pessimistic scenario. MAIN RESULTS AND THE ROLE OF CHANCE: CLBR had a declining trend with female age over six FET cycles (Groups 1-5, respectively: optimistic: 96.9%, 96.6%, 91.4%, 67.3%, and 11.7%; conservative: 87.3%, 85.0%, 74.0%, 41.3%, and 7.5%), requiring more FET cycles to achieve a success rate of at least 50% (Groups 1-5, respectively: optimistic: 2, 2, 2, 4, and >6 cycles; conservative: 2, 2, 2, >,6 and >6 cycles). CLBR showed an increasing trend with the number of oocytes retrieved (Groups 1-5, respectively: optimistic: 93.8%, 94.3%, 95.8%, 96.0%, and 95.6%; conservative: 66.2%, 78.3%, 85.6%, 88.9%, and 91.0%). All groups needed the same number of FET cycles to achieve a success rate of at least 50% (Groups 1-5, respectively: optimistic: 2, 2, 2, 2, and 2 cycles; conservative: 2, 2, 2, 2, and 2 cycles). Furthermore, the CLBR within six FET cycles had an increasing trend with AFC number (Groups 1-4, respectively: optimistic: 89.2%, 94.8%, 95.9%, and 96.3%; conservative: 67.4%, 78.2%, 83.9%, and 88.1%), with all four groups achieving a success rate of at least 50% by the second FET cycle. LIMITATIONS REASONS FOR CAUTION: The current research is limited by its retrospective design and single-centre nature, which may restrict the generalizability of our findings. WIDER IMPLICATIONS OF THE FINDINGS: This work describes two models (the Kaplan-Meier analysis and the competing risk method) to evaluate the clinical outcome of patients using PPOS-related protocols, which are especially useful for patients of advanced age or those with diminished ovarian reserve. Our findings encourage patients below 45 years old, especially younger than 40 years, and patients with lower AFCs and fewer retrieved oocytes to try this new protocol. Moreover, this study demonstrates the degree of improvement in the CLBR within six FET cycles for patients with different clinical characteristics, providing a valuable point of reference to determine whether to continue ART after a transfer failure. STUDY FUNDING/COMPETING INTERESTS: The study was supported by grants from the National Natural Science Foundation of China (82071603 to L.W., 82001502 to Y.L.). There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Water-Responsive Self-Contractive Silk-Based Skin Anti-Aging Tensioners with Customizable Biofunctions.

Skin anti-aging treatments have become increasingly popular. Currently, the prevalent treatment method involves implanting skin tension regulation threads (skin lifting threads) under the skin, and radiofrequency treatments. In this study, inspired by the natural supercontraction of spider silk, the molecular structure of silk fibroin fibers is modulated into an oriented configuration. This modification endows silk proteins with water-responsive self-contraction capabilities, leading to the development of innovative self-contracting silk-based skin tensioners (SSSTs). To align with clinical requirements, skin tension regulation materials are functionalized by testing for their self-contraction, near-infrared laser heating function, and bacteriostatic properties. The SSSTs exhibited remarkable self-contraction properties, drug-loading and sustained-release capabilities, notable antibacterial effects, controllable degradation, and good biocompatibility. Moreover, the near-infrared light heating function effectively increased subcutaneous temperature, demonstrating its potential for enhancing and prolonging skin lifting effects. Therefore, SSSTs can be applied for skin tension regulation to improve and delay skin aging. The results may pave the way for novel strategies in skin rejuvenation, with broad implications for the field of skin anti-aging.

Tissue/Organ Adaptable Bioelectronic Silk-Based Implants.

Implantable bioelectronic devices, designed for both monitoring and modulating living organisms, require functional and biological adaptability. Pure silk is innovatively employed, which is known for its excellent biocompatibility, to engineer water-triggered, geometrically reconfigurable membranes, on which functions can be integrated by Micro Electro Mechanical System (MEMS) techniques and specially functionalized silk. These devices can undergo programmed shape deformations within 10 min once triggered by water, and thus establishing stable bioelectronic interfaces with natively fitted geometries. As a testament to the applicability of this approach, a twining peripheral nerve electrode is designed, fabricated, and rigorously tested, demonstrating its efficacy in nerve modulation while ensuring biocompatibility for successful implantation.

The impact of online office on social anxiety among primary and secondary school teachers-Considering online social support and work intensity.

The COVID-19 has had a major impact on the global education system. In order to ensure the normal implementation of education courses, governments and education departments around the world have taken corresponding emergency measures. Based on data from 384 validated questionnaires, this study explored the effects of teleworking practices, work intensity, and online social support on social anxiety among primary and secondary school teachers. The results found that teleworking was more likely to cause social anxiety among teachers, while work intensity could promote social anxiety and online social support could reduce the probability of social anxiety. Work intensity can weaken the influence of partner support on social anxiety. Moreover, the model path coefficients differed across work styles. Based on the results, this study proposes some policy recommendations in order to provide theoretical guidance for improving social anxiety among primary and secondary school teachers and promoting the quality of educational work.

Internet use and depression among Chinese older adults: The mediating effect of interpersonal relationship.

The number of elderly Internet users has increased significantly in the past few years, and the Internet has greatly changed the way that older adults access information and communicate. Generally, those who regularly use the Internet may expand their range of interpersonal interactions, which has positive effects on their health. Depression is the leading cause of ill health, and is closely related to people's mental health. We sought to investigate whether internet use help reduce depression level among older adults. Using data from the 2020 China Family Panel Studies (CFPS), a total of 4,714 respondents were included to explore the effects of Internet use on the elderly's depression levels in China, along with the mediating role of interpersonal relationship in the above relationship. Regression results indicated that Internet use significantly reduced depression levels among the elderly. Further analysis showed that different Internet usage had different effects on depression among the elderly. Wechat chatting, video browsing, and online shopping were positively correlated with lower levels of depression. However, playing online games and online learning did not predict reduced levels of depression. Moreover, interpersonal relationship mediated the relationship between Internet use and depression levels. Internet use was associated with a higher level of interpersonal relationship, which in turn reduced depression levels in older adults. Regarding gender and regional differences, the coefficient of Internet use for urban older adults was significantly negative at 0.001 level, while it was not significant for rural older adults. A mediating effect of interpersonal relationship between Internet use and depression levels was only found for male elderly. To reduce the level of depression and promote mental health in the course of aging, Internet use and the improvement of interpersonal relationships merit special attention.

Drying kinetics and attributes of fructus aurantii processed by hot air thin-layer drying at different temperatures.

The primary objectives of this study were to evaluate the drying kinetics of Fructus Aurantii (FA), and to investigate how hot air drying at various temperatures affected the surface texture and sensory quality of the volatile fragrance components. The results were best simulated by the Overhults model, and use of scanning electron microscopy (SEM) and Heracles Neo ultra-fast gas phase electronic nose technology allowed for detection of changes in surface roughness and aromatic odors. The limonene content varied from 74.1% to 84.2% depending on the drying temperature, which ranged from 35°C to 75 °C. Furthermore, principal component analysis (PCA) revealed that the aromatic compound profile underwent considerable changes during the drying process. Overall, the present findings demonstrate that hot air thin-layer drying at 55 °C can significantly enhance the final quality of FA while preserving the taste properties and providing optimum medicinal and culinary characteristics.

Gut microbiota from essential tremor patients aggravates tremors in mice.

Background and objective: Essential tremor (ET) lacks effective treatments because its underlying mechanism is largely unknown, but may involve gut microbiota via the microbiome-gut-brain axis. We explored the effects of gut microbiota on ET in mice. Methods: Specific pathogen-free C57BL/6J mice were gavaged with stools from ET patients or matched healthy individuals. After 3 weeks of gavaging, behavioral tests were performed on all mice. Next, each mouse was injected with harmaline to induce tremors. The tremor duration was recorded; the tremor score was estimated every 30 min. Behavioral tests were repeated after modeling. Intestinal tissues and fecal samples of the mice were examined using histology and 16Sr DNA sequencing, respectively. Results: Compared with mice receiving microbiota from healthy controls, mice receiving fecal suspensions from ET patients showed worse performance in the pre-modeling behavioral tests. After modeling, ET-group mice showed significantly greater tremor scores, longer tremor duration, and worse motor performance. They also had significantly lower body weight and lower fecal pellet count. Pathological scoring revealed more severe intestinal lesions in ET-group mice. The 16S rDNA sequencing data revealed significant differences in microbiota indices, and a correlation between these indices and tremors in mice. Functional predictions indicated that the abundance of GABA-related enzymes was altered in ET-group mice. Conclusion: Mice transplanted with gut microbiota from ET patients showed worse performance in behavioral tests. After modeling, ET-group mice presented longer tremor duration, higher tremor score, and worse motor performance. This study provides evidence for gut microbiota dysbiosis that may affect the pathogenesis of ET.

A delayed ovulation of progestin-primed ovarian stimulation (PPOS) by downregulating the LHCGR/PGR pathway.

Progestin-primed ovarian stimulation (PPOS) is a new ovulation stimulation protocol, and its role in ovulation and regulatory mechanism is unclear. The clinical PPOS protocol was simulated in mice. The ovulated oocytes, estradiol, progesterone, and luteinizing hormone (LH) levels were analyzed at different hours after trigger. mRNA extraction and real-time PCR, hematoxylin and eosin staining, and immunofluorescence of ovaries were used to explore the involved signaling pathways. The PPOS group had a delayed ovulation at 12.5 h after trigger. Its suppressed LH level reduced the expression of luteinizing hormone/choriogonadotropin receptor (LHCGR) on the preovulatory follicles before trigger and significantly decreased the following progesterone synthesis, blood progesterone level, and progesterone receptor (PGR) expression within 4-6 h after trigger. Furthermore, the important ovulatory genes regulated by PGR including ADAMTS-1, VEGF-A, and EDN2 were downregulated, ultimately delaying the ovulation. PPOS suppresses the LH level before trigger and decreases the synthesis of progesterone after trigger, thus delaying the ovulation by downregulating the LHCGR-PGR pathway.

Correlations between flavor and fermentation days and changes in quality-related physiochemical characteristics of fermented Aurantii Fructus.

The effects of different fermentation times (0, 1, 2, 3, 4, and 5 days) on the physicochemical properties and flavor components of fermented Aurantii Fructus (FAF) were evaluated. Component analysis identified 66 compounds in positive ion mode and 32 compounds in negative ion mode. Flash GC e-nose results showed that propanal, (+)-limonene and n-nonanal may be the flavor characteristic components that distinguish FAF with different fermentation days. Furthermore, we found that the change of total flavonoid content was closely related to colony growth vitality. The total flavonoid content of FAF gradually decreased from 3rd day and then increased from 5th day (3rd day: 0.766 ± 0.123 mg/100 g; 4th day: 0.464 ± 0.001 mg/100 g; 5th day: 0.850 ± 0.192 mg/100 g). Finally, according to antioxidant activity correlation analysis, meranzin, (+)-limonene and total flavonoids were found to be the key substances affecting the fermentation days of FAF. Overall, the optimal fermentation time for FAF was 4 days.

Earlier second polar body transfer and further mitochondrial carryover removal for potential mitochondrial replacement therapy.

The second polar body (PB2) transfer in assisted reproductive technology is regarded as the most promising mitochondrial replacement scheme for preventing the mitochondrial disease inheritance owing to its less mitochondrial carryover and stronger operability. However, the mitochondrial carryover was still detectable in the reconstructed oocyte in conventional second polar body transfer scheme. Moreover, the delayed operating time would increase the second polar body DNA damage. In this study, we established a spindle-protrusion-retained second polar body separation technique, which allowed us to perform earlier second polar body transfer to avoid DNA damage accumulation. We could also locate the fusion site after the transfer through the spindle protrusion. Then, we further eliminated the mitochondrial carryover in the reconstructed oocytes through a physically based residue removal method. The results showed that our scheme could produce a nearly normal proportion of normal-karyotype blastocysts with further reduced mitochondrial carryover, both in mice and humans. Additionally, we also obtained mouse embryonic stem cells and healthy live-born mice with almost undetectable mitochondrial carryover. These findings indicate that our improvement in the second polar body transfer is conducive to the development and further mitochondria carryover elimination of reconstructed embryos, which provides a valuable choice for future clinical applications of mitochondrial replacement.

Comparative Analysis of the Chalcone-Flavanone Isomerase Genes in Six Citrus Species and Their Expression Analysis in Sweet Orange (Citrus sinensis).

Citrus fruit contains rich nutrients which is edible and of officinal value. Citrus flavanones are widely used in the treatment of cardiovascular and other diseases, and they are a foundational material of Chinese medicine. The chalcone-flavanone isomerase (CHI) plays a key role in flavanone synthesis. Therefore, we comprehensively analyzed CHI genes in Citrus species. Here, thirty CHI genes were identified for the first time in six Citrus species, which were divided into CHI and FAP groups. Evolutionary analysis showed that CHI gene members were highly conserved and were an ancient family. All CsCHI genes showed the highest expression level after the second physiological fruit-falling period in C. sinensis. CsCHI1 and CsCHI3 were highly expressed at 50 days after the flowering (DAF) stage in albedo. The expression of CsFAP2 and CsCHI3 genes at the 50 DAF stage was 16.5 and 24.3 times higher than that at the 220 DAF stage, respectively. The expression of CsCHI1, CsCHI3, and CsFAP2 genes in the peel was higher than that in the pulp, especially in common sweet orange. The CsCHI3 gene maintained a high expression level in the epicarp and juice sac at all periods. The members of CHIs interacted with chalcone synthase (CHS), flavonol synthase/flavanone 3-hydroxylase (FLS) and naringenin, and 2-oxoglutarate 3-dioxygenase (F3H) to form heterodimers, which might together play a regulatory role and participate in the flavonoid pathway. This study will provide the basis for the selection of flavonoids in plant tissues and periods and fundamental information for further functional studies.

Long-term efficacy and safety of cap-assisted endoscopic sclerotherapy with long injection needle for internal hemorrhoids.

BACKGROUND: Hemorrhoids are a common anal condition and can afflict an individual at any age. Epidemiological survey results in China show that the prevalence of anorectal diseases is as high as 50.1% among which 98.08% of patients have hemorrhoid symptoms. AIM: To assess long-term efficacy and safety of cap-assisted endoscopic sclerotherapy (CAES) with long injection needle for internal hemorrhoids. METHODS: This study was retrospective. Data from patients with symptomatic internal hemorrhoids treated with CAES using endoscopic long injection needle from April 2016 to December 2019 were collected. Patients were telephoned and followed at two time points, December 2020 and 2021, to evaluate the improvements in symptoms, complications, recurrence, and satisfaction. RESULTS: Two hundreds and one patients with internal hemorrhoids underwent CAES with the long needle. The first median follow-up was performed 33 mo post-operatively. Symptoms improved in 87.5% of patients after the first CAES. Efficacy did not decrease with treatment time extension. Fifty-four patients underwent colonoscopy after the first CAES treatment of which 21 underwent CAES again, and 4 underwent hemorrhoidectomy. At the first follow-up, 62.7% of patients had both improved hemorrhoid grades and symptoms, and 27.4% had a significant improvement in both parameters. At the second follow-up, 61.7% of the patients showed satisfactory improvement in their hemorrhoid grade and symptoms when compared with pre-surgery values. 90% of patients reported CAES was painless, and 85% were satisfied/very satisfied with CAES treatment outcomes. CONCLUSION: The present study based on the largest sample size reported the long-term follow-up of the treatment for internal hemorrhoid with the CAES using endoscopic long injection needle. Our findings demonstrate that CAES should be a micro-invasive endoscopic technology yields satisfactory long-term efficacy and safety.