Effects of vitrification on mitochondrial ultrastructure and membrane potential and its distribution in mouse oocytes.

BACKGROUND: Vitrification is commonly used for in vitro fertilization and has significant impact on gametes. OBJECTIVE: To investigate changes in ultrastructure, membrane potential and distribution of mitochondria in mouse oocytes after vitrification. MATERIALS AND METHODS: Mouse oocytes were divided into three groups: one group as fresh control, one group for the toxicity test (treated with cryoprotectant but without vitrification), and the other for vitrification. RESULTS: Most mitochondria in oocytes were damaged after cooling and warming, being rough and fuzzy in appearance, even swollen and broken. The membrane potential of the toxicity test group and the vitrification group was 0.320 +/-0.030 and 0.244 +/- 0.038, respectively, in comparison to the fresh group (0.398 +/- 0.043). The membrane potential of the vitrified oocytes was significantly lower than fresh oocytes and the toxicity test oocytes (P % 0.05), but there was no significant difference between fresh oocytes and the toxicity test oocytes (P > 0.05). Mitochondria in fresh oocytes were denser and strained stronger, with 59.5> distributed homogeneously and 36.4> polarized. The majority of mitochondria in the toxicity-tested oocytes were clustered (69.3>) and only a small portion were distributed homogeneously (19.6>), while mitochondria in vitrified oocytes were clustered (56.3>) and deficient (24.4>), and their fluorescent staining was weak and blurred. There was a significant disruption in mitochondrial function after vitrification. CONCLUSION: Vitrification alters the ultrastructure, membrane potential and distribution of mitochondria in oocytes, most likely caused by toxicity and mechanical injury. Doi.org/10.54680/fr24510110212.

[A quantitative MRI comparative study of imaging markers for cerebral small vessel disease in the middle-aged and elderly patients with and without hypertension].

Objective: To explore the difference of MRI markers of small cerebral vascular disease in middle-aged and elderly patients with hypertension and non-hypertension. Methods: A cross-sectional study. The clinical data of 316 patients who underwent head MRI with susceptibility weighted imaging scans at the Affiliated Zhongda Hospital of Southeast University from November 2013 to August 2022 were retrospectively analyzed, including 190 males and 126 females, with the age of (71.6±8.9)years. According to the history of hypertension, the patients were divided into hypertension group(n=259) and the non-hypertension group(n=57). The patients in the non-hypertension group were further divided into abnormal blood pressure group on admission (n=19) and normal blood pressure group on admission (n=38). The imaging features of different CSVD dimensions in the patient's images were quantified or graded and compared between hypertensive and non-hypertensive patient groups. Deep learning methods were employed to segment white matter lesions, and voxel-wise analysis was conducted to investigate the differences in whole-brain white matter lesion probability between patients in both groups. Spearman correlation analysis was used to analyze the correlation between hypertension and small cerebral vascular disease. Results: Compared with the non-hypertensive group, the cerebral microhemorrhage count, deep microhemorrhage count, basal ganglia level lacunae count, perivascular space (EPVS) grade of hemioval center level and EPVS grade of basal ganglia level were higher in the hypertensive group (all P<0.05). The cerebral microhemorrhage count [3.0(1.0, 15.0) vs 1.0 (0, 4.2)], deep microhemorrhage count [1.0 (0, 7.0) vs 1.0 (0, 4.2)] and EPVS classification at basal ganglium level [2.0(1.0, 3.0) vs 1.0(1.0, 2.0)] in the group with history of hypertension were higher than those in the group with normal blood pressure at admission (all P<0.05). The EPVS grade at the central level of the semiovale in the hypertension group was higher than that of the group with normal blood pressure at admission [2.0(1.0, 2.0) vs 1.0 (1.0, 2.0)], and also higher than that of the group with abnormal blood pressure at admission [2.0(1.0, 2.0) vs 1.0(1.0, 2.0)](both P<0.05). Voxel-by-voxel analysis showed no significant difference in the probability of white matter lesions in the whole brain between patients with and without a history of hypertension, but patients with a history of hypertension showed more extensive para-ventricular white matter hypersignaling than those without a history of hypertension. Spearman correlation analysis showed that hypertension grade was correlated with the number of microbleeding lesions in depth (r=0.149), the number of lacunae lesions in the center of the hemioval (r=0.209), and the number of lacunae lesions in the basal ganglia (r=0.204) (all P<0.05). Conclusions: Chronic hypertension can affect different dimensions of small vessel disease imaging, primarily manifested in the increases of deep microbleed counts and the EPVS grade.

[The long-term efficacy of metformin in megestrol acetate-based fertility-sparing treatment for patients with endometrial atypical hyperplasia and endometrioid endometrial cancer].

Objective: To assess the long-term efficacy of metformin in megestrol acetate (MA)-based fertility-sparing treatment for patients with endometrial atypical hyperplasia (EAH) and endometrioid endometrial cancer (EEC). Methods: The randomized controlled trail study was conducted from October 2013 to October 2017 in the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China. Patients with EAH or EEC were firstly stratified according to pathology, and randomized to receive MA (160 mg orally, daily) plus metformin (500 mg orally, three times a day) or MA (160 mg orally, daily). Baseline data between two groups of patients were compared. Estimates of time to complete remission (CR) and recurrence-free survival (RFS) were calculated using the Kaplan-Meier method. Cox proportional-hazards regression model was used to estimate hazard ratios (HR) of related factors for recurrence-free survival. Quantitative data were represented by M (Q1, Q3). Results: A total of 150 patients were included, and 76 patients were allocated to receive MA plus metformin with the age of 32.5 (28.0, 36.0), while 74 patients received MA alone with the age of 32.0 (28.0, 36.0). By the end of follow-up period, 96.7% (n=145) of patients achieved complete remission, with a median follow-up time of 57.7 (26.7, 70.5) months. The median CR time for the MA plus metformin group and the MA alone group were 6.3 (3.5, 8.3) months and 6.8 (4.0, 9.3) months, respectively (P=0.193), with 2-year cumulative CR rate of 98.6% and 98.5%, respectively (P=0.879). The median time of RFS was 28.1 (12.5, 57.3) months for the MA plus metformin group and 33.3 (14.1, 62.5) months for the MA alone group (P=0.213), with a cumulative RFS rate of 61.9% and 65.8%, respectively (P=0.560). In the subgroup of non-obese (body mass index<28 kg/m2) patients with EAH, the median RFS times were 25.7 (7.6, 60.3) months and 47.3 (17.5, 64.8) months for the MA plus metformin group and the MA alone group, respectively (P=0.033), with a cumulative RFS rate of 57.5% and 80.6%, respectively (P=0.029). According to Cox proportional hazards regression analysis, undergoing assisted reproductive treatment (HR=2.358, 95%CI: 1.069-5.204, P=0.034) was identified as an independent risk factor for recurrence-free survival after complete remission of endometrial lesions. Conclusion: The long-term follow-up outcome indicates that there is no significant difference in CR time and RFS time between MA plus metformin therapy and MA alone therapy for patients with EAH or EEC.

[Comparison of the clinical efficacy of all-inside arthroscopic lateral ligament augmentation procedure and Broström procedure for the treatment of chronic lateral rotational ankle instability].

Objective: To compare the clinical efficacy of patients with chronic lateral rotational ankle instability(CLRAI) after all-inside arthroscopic lateral ligament augmentation procedure and Broström procedure. Methods: This is a retrospective cohort study. The clinical and imaging data of 106 CLRAI patients were collected at the Xuzhou Central Hospital from January 2021 to December 2022. The patients included 55 males and 51 females with an age of (32.6±8.2) years (range: 16 to 50 years). All patients were treated under all-inside arthroscopic, and were divided into Broström-Gould surgery group (n=54) and Broström surgery group (n=52) according to different ligament repair methods. At 3, 6, and 12 months after surgery, ankle inversion stress tests and anterior drawer tests were used to examine the stability of the ankle joint and observe gait. The American Orthopedic Foot and Ankle Society ankle hindfoot scale (AOFAS-AH) and Karlsson ankle function score (KAFS) were used to assess ankle function; Tegner score was used to assess the patient's level of exercise; the foot and ankle outcome score(FAOS)(including score of symptoms,pain,function, daily living,function, sports and recreational activities (sport); quality of life (QOL) was used to assess the patient's daily activity ability. Comparisons of data were made using independent sample t test, repeated measures analysis of variance, LSD-t multiple comparison method, χ2 test or Mann-Whitney U test. Results: All operations were successfully accomplished. All incisions healed by first intention, without evidence of postoperative complications of implant rejection, ligation reaction, and nerve and vessel injury. All patients were followed up at 3, 6, and 12 months after surgery. Ankle varus stress test and anterior drawer test were negative. No evidence supporting lateral ankle instability was obtained. All patients eventually regained normal gait. No patients underwent revision surgery. Repeated measurement analysis of variance showed that AOFAS-AH, Tegner, KAFS and FAOS scores in the Brostrom-Gould group and the Brostrom group were significantly higher than those before surgery (P<0.01). The change trends of Tegner score and FAOS-sport score were significantly different between the two groups (F=18.839, P<0.01; F=8.169, P=0.005). Multiple comparisons revealed that at 3-, 6-and 12-month follow-up, the Tegner scores (3 months: 3.7±0.5 vs. 3.3±0.5, t=-3.980, P<0.01; 6 months: 4.4±0.6 vs. 3.8±0.7, t=-4.792,P<0.01; 12 months: 5.8±0.9 vs. 5.1±1.0, t=-3.889,P<0.01), sport scores (3 months: 82.5±3.7 vs. 79.3±3.8, LSD-t=-4.316, P<0.01; 6 months: 88.5±4.9 vs. 85.7±3.8, LSD-t=-3.312,P=0.001;12 months: 90.1±4.3 vs. 88.2±5.1, LSD-t=-2.112,P=0.037) in the Broström-Gould surgery group were higher than those in the Broström surgery group, with statistical significances. Conclusions: Both Broström-Gould and Broström procedures under all-inside arthroscopic can make ankle stability and improve ankle function in the treatment of CLRAI. However, the former maybe shorten the time to return to exercise and achieve higher motor function.

[Discussion and evaluation of several issues concerning the classification of impacted third molars].

The classification of impacted third molars is important in clinical diagnosis and surgical selection. At present, the common clinical impacted third molar classification is different in the maxillary and mandibular classification criteria. Through the review and analysis of various classification methods of impacted third molar, this paper proposes a more comprehensive and unified classification proposal for the classification of impacted third molar, in order to form a widely accepted application classification method.

[Deciphering the fate of granulation tissue in the human inflammatory alveolar microenvironment using single-cell RNA sequencing].

Objective: To investigate the cellular composition and heterogeneity of granulation tissue in human alveolar sockets and construct single-cell transcriptomic maps to elucidate the potential outcomes of natural resolution in the inflammatory microenvironment. Methods: Granulation tissues from 12 alveolar sockets undergoing tooth extraction due to periodontitis and scheduled for delayed site preservation or autologous tooth transplantation were collected in the Department of Oral and Maxillofacial Surgery, School of stomatology Fourth Military Medical University from September 2022 to August 2023. This study used HE staining and single-cell RNA sequencing (scRNA-seq) to observe the cellular composition and morphological changes of different types of granulation tissues. This approach enabled us to identify cellular sequence variations in the inflammatory dental alveolar granulation tissue within specific microenvironments, construct single-cell atlases for different types of human dental alveolar granulation tissues, and explore the spatiotemporal patterns of cell type distribution and key gene changes during the resolution process of inflammatory granulation tissue. Results: HE staining revealed extensive infiltration of inflammatory cells in the dental alveolar inflammatory granulation tissue. After allowing the inflammatory granulation tissue to naturally resolve for three weeks, its histological morphology was essentially consistent with that of reparative granulation tissue. ScRNA-seq captured a total of 20 448 cells from granulation tissues, and the gene expression quantification analysis revealed total gene counts of 33 835 for inflammatory granulation tissue, 36 058 for naturally resolved granulation tissue, and 34 719 for reparative granulation tissue. At the single-cell level, granulation tissue was annotated into ten cell subgroups, including vascular endothelial cells, mesenchymal stem cells, and fibroblasts. Differences were observed in the proportion of cell compositions between inflammatory and naturally resolved granulation tissues. Pseudo-temporal analysis illustrated two main outcomes in tissue resolution and healing, involving immune responses and angiogenesis. Among these, genes associated with inflammation regulation and immune response, such as Igbp5, Zfp36, and Hspa1b, as well as genes involved in extracellular matrix secretion and the formation of vessels and fibers such as Timp3, Postn, and Rgs5, showed significant differences in expression between the two types of granulation tissues. Conclusions: Inflammatory granulation tissue exhibits heterogeneity in cell composition, gene expression, and biological functions compared to naturally resolved granulation tissue. When the inflammatory granulation tissue in the alveolar socket is left undisturbed to undergo natural resolution, it displays a cellular composition similar to that of reparative granulation tissue at both the histological and single-cell levels. Moreover, it modulates the inflammatory response and the healing process through immune reactions and tissue remodeling.

[Strolling through the glorious years of Alveolar Surgery, bravely stepping onto the path of practice and innovation].

This article summarizes and organizes relevant publications in journals, along with a review of medical history, systematically summarizing the development process of dental alveolar surgery in China. The initial establishment phase (1935-1952) marked the starting point of Chinese Alveolar Surgery. Despite the impact of wars, it laid the foundation for subsequent research and practice. During the early development phase (1953-1966), the "Chinese Journal of Stomatology" was founded, which promoted the development of Alveolar Surgery. Research focused on tooth extraction methods and complications. Tooth Transplantation and Preprosthetic Surgery gradually began to take off. The stagnant phase (1967-1977) occurred due to the interruption of international exchanges, leading to an almost complete halt in the development of Alveolar Surgery. Entering the rapid catch-up phase (1978-1985), Alveolar Surgery scholars in China began striving to overcome the stagnation of the previous decade. While some progress was made, no significant innovative achievements emerged. In the scientific development phase (1986-2010), clinical research, basic experiments, and paper writing in modern Chinese Alveolar Surgery began to adhere to scientific standards with the rise of experimental medicine. The exploration and innovation stage (2011-2023) is the current development phase, during which Chinese Aveolar Surgery has reached its peak, making substantial progress in technology, clinical practices, and basic research, gradually reaching or even surpassing international advanced levels. Looking back at the development history in China, we can find the wisdom and hard work of the older generation of Alveolar Surgery scholars. However, contemporary challenges and issues, such as standardizing technology, promoting clinical practices, and talent cultivation, need to be addressed by present-day Alveolar Surgery professionals as they forge ahead.

Involvement of Src tyrosine kinase and protein kinase C in the expression of macrophage migration inhibitory factor induced by H2O2 in HL-1 mouse cardiac muscle cells

Macrophage migration inhibitory factor (MIF), a pleiotropic cytokine, plays an important role in the pathogenesis of atrial fibrillation; however, the upstream regulation of MIF in atrial myocytes remains unclear. In the present study, we investigated whether and how MIF is regulated in response to the renin-angiotensin system and oxidative stress in atrium myocytes (HL-1 cells). MIF protein and mRNA levels in HL-1 cells were assayed using immunofluorescence, real-time PCR, and Western blot. The result indicated that MIF was expressed in the cytoplasm of HL-1 cells. Hydrogen peroxide (H2O2), but not angiotensin II, stimulated MIF expression in HL-1 cells. H2O2-induced MIF protein and gene levels increased in a dose-dependent manner and were completely abolished in the presence of catalase. H2O2-induced MIF production was completely inhibited by tyrosine kinase inhibitors genistein and PP1, as well as by protein kinase C (PKC) inhibitor GF109203X, suggesting that redox-sensitive MIF production is mediated through tyrosine kinase and PKC-dependent mechanisms in HL-1 cells. These results suggest that MIF is upregulated by HL-1 cells in response to redox stress, probably by the activation of Src and PKC.