This study primarily investigates the effect of Liuwei Dihuang Pills on the activation and proliferation of female germline stem cells(FGSCs) in the ovaries and cortex of mice with premature ovarian failure(POF), and how it improves ovarian function. ICR mice were randomly divided into the control group, model group, Liuwei Dihuang Pills group, Liuwei Dihuang Pills double-dose group, and estradiol valerate group. A mouse model of POF was established by intraperitoneal injection of cyclophosphamide. After successful modeling, the mice were treated with Liuwei Dihuang Pills or estradiol valerate for 28 days. Vaginal smears were prepared to observe the estrous cycle and body weight. After the last administration, mice were sacrificed and sampled. Serum levels of estradiol(E_2), follicle-stimulating hormone(FSH), luteinizing hormone(LH), and anti-Müllerian hormone(AMH) were measured by enzyme-linked immunosorbent assay(ELISA). Hematoxylin-eosin(HE) staining was used to observe ovarian morphology and to count follicles at all stages to evaluate ovarian function. Immunohistochemistry was used to detect the expression of mouse vasa homolog(MVH), a marker of ovarian FGSCs. Immunofluorescence staining, using co-labeling of MVH and proliferating cell nuclear antigen(PCNA), was used to detect the expression and localization of specific markers of FGSCs. Western blot was employed to assess the protein expression of MVH, octamer-binding transcription factor 4(Oct4), and PCNA in the ovaries. The results showed that compared with the control group, the model group exhibited disordered estrous cycles, decreased ovarian index, increased atretic follicles, and a reduced number of follicles at all stages. FSH and LH levels were significantly elevated, while AMH and E_2 levels were significantly reduced, indicating the success of the model. After treatment with Liuwei Dihuang Pills or estradiol valerate, hormone levels improved, the number of atretic follicles decreased, and the number of follicles at all stages increased. MVH marker protein and PCNA proliferative protein expression in ovarian tissue also increased. These results suggest that Liuwei Dihuang Pills regulate estrous cycles and hormone disorders in POF mice, promote the proliferation of FGSCs, improve follicular development in POF mice, and enhance ovarian function.
Animals ; Female ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Cell Proliferation/drug effects* ; Mice, Inbred ICR ; Ovary/cytology* ; Primary Ovarian Insufficiency/genetics* ; Follicle Stimulating Hormone/metabolism* ; Humans ; Anti-Mullerian Hormone/blood* ; Antineoplastic Agents/adverse effects* ; Luteinizing Hormone/metabolism* ; Cyclophosphamide/adverse effects*

Bombesin receptor subtype-3 (BRS3) is an orphan G protein-coupled receptor (GPCR) that plays critical roles in energy homeostasis, glucose metabolism, and insulin secretion. Recent structural studies have elucidated BRS3 signaling mechanisms using synthetic ligands, including BA1 and MK-5046. However, the molecular basis of BRS3 activation by bioactive natural compounds and their derivatives, particularly those derived from traditional Chinese medicine, remains unclear. Here, we present high-resolution cryogenic electron microscopy (cryo-EM) structures of the human BRS3-G_q complex in both unliganded and active states bound by two herb-derived compounds (DSO-5a and oridonin), at resolutions of 2.9, 2.8, and 2.9 Å, respectively. These structures display distinct ligand recognition patterns between DSO-5a and oridonin. Although both compounds bind to the orthosteric pocket, they differentially engage the interaction network of BRS3, as demonstrated by mutagenesis studies assessing calcium mobilization and inositol phosphate 1 (IP1) accumulation. These findings enhance our understanding of BRS3 activation and provide valuable insights into the development of small-molecule BRS3 modulators with therapeutic potential.

Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.

Objective:Discipline construction is the key and effective method for the high-quality development of hospitals. This study summarized the practical and valuable experience of discipline construction with the support of the ″Sanming Project of Medicine″ in Shenzhen, to provide a worthy reference for similar projects and hospital specialty development.Methods:Taking the introduction of the hematology-oncology team of the Canadian Children′s Hospital as an example, we analyzed the background, the process, and the effectiveness of discipline construction of hematology and oncology of Shenzhen Children′s Hospital under the support of the ″Sanming Project of Medicine″, and explored the common rules for rapid specialty development.Results:Through the implementation of two levels supporting policies from cities and hospitals and quantifiable and verifiable annual progressive program, the adoption of the international advanced medical management mode and concepts, and the transformation and application of cutting-edge clinical technologies, we successfully built the largest pediatric hematologic oncology diagnosis and treatment center in South China and the largest pediatric hematopoietic stem cell transplant center in China. By executing a multi-level talent developing program, the number of specialists with MD and PhD degrees increased by 6.5 times, and 23 doctors and nurses were trained abroad, leading to the development of provincial key clinical specialties and municipal key scientific research platforms.Conclusions:This project concluded the beneficial experience of introducing high-level overseas medical teams to boost discipline construction, including internationally advanced medical concepts and management modes, tiered talent cultivation schemes, the construction of clinical and basic research platforms, etc., providing references for the rapid improvement of discipline construction in terms of health care for other hospitals in Shenzhen and similar cities.

Objective:To investigate the safety and efficacy of low negative pressure fat suction combined with bipolar radiofrequency in repairing facial fat overfilling.Methods:A total of 29 patients with facial fat overfilling underwent low negative pressure fat suction combined with bipolar radiofrequency between February 2022 and October 2023 in Changsha My Like Medical Cosmetology Hospital. All patients were female. The mean age was 35.7 years (range, 25-51 years) and the mean body mass index was 20.3 (range, 16.8-24.3 kg/cm 2). The mean time after autologous fat transplantation was 53.4 months (range, 19-96 months). Postoperative patient satisfaction surveys were conducted, and 2 independent doctors evaluated clinical effect preoperative and postoperative photographs at 3-6 months postoperatively. Results:There were 29 cases (100%) underwent liposuction in the pre-zygomatic region, and the amount of fat was 4.1-12.6 (7.95±1.85) ml. There were 4 cases (13.8%) underwent liposuction in the forehead, and the amount of fat was 1.8-5.2 (2.75±0.50) ml. There were 7 cases (24.1%) underwent liposuction in the temporal region, and the amount of fat was 2.8-6.5 (3.86±1.07) ml. There were 8 cases (27.6%) underwent liposuction in the cheek, and the amount of fat was 5.2-10.5 (7.25±2.12) ml. There were 18 cases (62.1%) underwent liposuction in the perioral region, and the amount of fat was 3.7-9.5 (6.33±1.28) ml. The energy delivered by bipolar radiofrequency was 3.3-10.2 (5.71±2.27) KJ. 82.8% of patients were satisfied with their postoperative effect (24/29 patients). 86.2% of doctors were satisfied with the postoperative effect (25/29 patients). Two out of 29 patients (6.9%) developed irregularity by liposuction.Conclusions:Low negative pressure liposuction combined with bipolar radiofrequency can effectively reduce the fat, narrow the tissue gap and improve facial sagging, which is an effective method for repairing facial fat overfilling.

Objective:To explore the effect and safety of mesenchymal stem cell exosome microneedle introduction in the treatment of melasma.Methods:Thirty cases of female patients with stable melasma in the Department of Dermatology, Changsha Meilai Medical Beauty Hospital, aged (36±5) years and with a disease duration of (42.4±20.7) months, from July 2021 to July 2022, were retrospectively included. According to Fitzpatrick skin typing, 23 cases of type Ⅲ and 7 cases of type Ⅳ were included. All patients were locally anesthetized with lidocaine cream for 30 min, and rolled with a 0.5 mm needle in a zigzag pattern with even force, in the order of the right cheek, the left cheek, the forehead, the nose, the mandible, and the upper lip. During the rolling process, 3 ml of MSC exosome medical liquid wound dressing was applied to the facial skin, and after it was fully absorbed, exosome was locally readministered in the area of melasma. Treatment ended with a slight redness at the site of application. 1 MSC exosome wound dressing was appllied as a cold compress for 15 min after treatment. Treatment was given once every 2 weeks for 6 consecutive sessions. All the patients were followed up at 4 and 12 weeks after the last session, and the area and severity index of melasma (MASI) were scored before and after the treatment, the clinical efficiency and patient satisfaction rate and the incidence of adverse reactions were also counted.Results:At 4 and 12 weeks after the end of treatment, the skin color of all 30 patients was brighter than that before treatment, and no recurrence of melasma symptoms seen. At 12 weeks after the end of treatment, the decrease rate of MASI score was 66.1%, among which the decrease rate of MASI score in patients with type Ⅲ melasma was 63.9%, and the decrease rate of MASI score in patients with type Ⅳ melasma was 63.9%. Among the 30 patients, 1 case was cured, 25 cases showed obvious improved, 4 cases were improved, and no cases were ineffective, with an effective rate of 86.7% (26/30). Five patients were very satisfied, 18 patients were satisfied, 6 patients were generally satisfied, and 1 patient was dissatisfied; the patient satisfaction rate was 76.7% (23/30). No serious adverse reactions occurred in all patients.Conclusions:MSC exosome microneedle introduction is safe and effective in the treatment of melasma without serious adverse reactions.

Innate and adaptive immune responses initiated by infection depend on recognition and control of pathogens by macrophages,dendritic cells,T cells and so on.Notch signaling pathway is a highly conserved pathway,which is activated by interac-tion of receptors and ligands,thus coordinating important life processes of cells.At present,it has been confirmed that Notch signaling pathway is involved in development,differentiation,maturation and activation of many kinds of immune cells,and plays an important role in infectious diseases.In this review,we mainly focus on the role of Notch signaling pathway in immune regulation during different pathogens infection and its interaction with other signaling pathways.Additionally,therapeutic methods and challenges of developing Notch signaling as a target in infectious diseases are also discussed.

Objective To investigate the role of bone marrow mesenchymal stem cells derived from diabetic mice and their paracrine roles in inducing insulin resistance(IR).Methods The mouse model of diabetes mellitus was established,bone marrow mesenchymal stem cells(BMSC)were extracted and cultured,and the culture supernatant(M-BMSC-CS)was collected.(1)Cell experiment:HepG2 hepatocytes were divided into normal low-glycemic culture group[cultured with low-glycemic DMEM(5.55 mmol·L-1)],M-BMSC-CS experimental group(M-BMSC-CS 75 μL),and high-glycemic and high-lipid control group(given 25 mmol·L-1 high-glycemic DMEM+0.25 mmol·L-1 palmitic acid);(2)Animal experiments:Mice were divided into normal mice group(0.9％NaCl by intraperitoneal injection)and M-BMSC-CS-m group(M-BMSC-CS by intraperitoneal injection of normal mice(injection dose 0.2 mL/10 g)].Glucose intake was measured by glucose oxidase method.The fluorescence intensity of Glut2 protein was detected by immunofluorescence.The expression of insulin signaling pathway protein was detected by Western blot.Test oral glucose tolerance(OGTT)and insulin tolerance(ITT).Results The glucose intakes of the normal low-glucose culture group,the M-BMSC-CS experimental group and the high-glucose and high-lipid control group were(2.96±0.05),(1.64±0.28)and(1.42±0.32)mmol·L-1,respectively;the fluorescence expressions of glucose transporter 2(Glut2)were 53.21±2.70,30.95±3.39 and 34.96±7.60,respectively;the protein expression levels of phosphorylated insulin receptor substrate 1-ser307(p-IRS-1ser307)were 0.46±0.21,1.09±0.24 and 0.91±0.16,respectively;phosphorylated protein kinase(p-AKT)protein expression levels were 0.94±0.05,0.59±0.06 and 0.53±0.05;Glut2 protein expression levels were 1.08±0.14,0.58±0.14 and 0.62±0.09,respectively.The above indexes in M-BMSC-CS experimental group were statistically significant compared with those in normal low-glycemic culture group(all P＜0.05).Fasting blood glucose levels in the normal group and M-BMSC-CS-m group were(5.23±0.57)and(9.30±1.14)mmol·L-1;p-AKT protein expression level were 1.27±0.21 and 0.51±0.19;Glut2 protein expression level were 1.17±0.17 and 0.79±0.09,respectively.The above indexes in M-BMSC-CS-m group were significantly different from those in normal mouse group(P＜0.05).Conclusion BMSC culture supernatant from diabetic mice induced insulin resistance of normal HepG2 hepatocytes in vitro and normal mice in vivo.

Objective To investigate the relationship between the injury and ferroptosis of bone marrow mesenchymal stem cells(BMSCs)induced by high glucose and high fat.Methods BMSCs were divided into normal group(5.50 mmol·L-1 glucose)and high glucose and high fat(HGHF)group(25.00 mmol·L-1 glucose+0.25 mmol·L-1 palmitic acid).Assessment of cellular aging via β-galactosidase staining;enzyme linked immunosorbent assay(ELISA)were used to detect tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)release levels;glutathione(GSH),malondialdehyde(MDA)and ferrous ion(Fe2+)detection kits were used to detect ferroptosis related indicators;Western blotting was used to detect the expression of ferroptosis related signaling pathway protein acyl-CoA synthetase long chain family member 4(ACS14)/arachidonate 15-lipoxygenase(ALOX15)/glutathione peroxidase 4(GPX4).Results The senescence rates of normal group HGHF group were(6.80±1.60)％and(13.00±1.58)％;the levels of TNF-α were(122.54±3.94)and(169.77±2.89)pg·mL-1;the levels of IL-10 were(155.16±3.97)and(105.15±7.30)pg·mL-1;GSH levels were 4.30±0.33 and 1.55±0.14;MDA levels were 2.94±0.10 and 5.84±0.10;Fe2+levels were 6.22±0.35 and 16.13±0.36;the relative expression levels of ACSL4 protein were 0.42±0.05 and 0.84±0.10;the relative ALOX15 protein were 0.61±0.25 and 1.06±0.11;the relative expression levels of GPX4 protein were 1.13±0.17 and 0.33±0.08,respectively.The above indexes in the HGHF group were significantly different from those in the normal group(all P＜0.05).Conclusion 25 mmol·L-1 glucose combined with 0.25 mmol·L-1 palmitic acid for 24 h can be used as a suitable condition to induce BMSCs injury.ferroptosis plays an important role in BMSCs injury induced by high glucose and high fat.