Dehydroepiandrosterone promotes ovarian angiogenesis and improves ovarian function in a rat model of premature ovarian insufficiency by up-regulating HIF-1α/VEGF signalling.

RESEARCH QUESTION: What impact does dehydroepiandrosterone (DHEA) have on ovarian angiogenesis and function in a rat model of with premature ovarian insufficiency (POI), and what are the potential mechanisms of action? DESIGN: DHEA was added to a culture of human microvascular endothelial cells (HMEC-1) to investigate its effects on cell proliferation, migration and tube formation. A rat model of POI was established by intraperitoneal injection of cyclophosphamide, followed by continuous oral administration of DHEA or vehicle for 28 days. Ovarian angiogenesis, follicular growth and granulosa cell survival in ovarian tissues were assessed through haematoxylin and eosin staining, immunohistochemistry and TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL). The effect of DHEA on the fertility of rats with POI was evaluated in pregnant animals. The expression levels of characteristic genes and proteins in the hypoxia-inducible factor (HIF)-1α/vascular endothelial growth factor (VEGF) pathway was determined using quantitative reverse transcription PCR and western blotting. RESULTS: In-vitro experiments revealed that DHEA stimulated the proliferation, migration and tube formation of HMEC-1. In in-vivo studies, DHEA treatment improved the disruption of the oestrous cycle and hormone imbalances in POI rats. Key genes in the HIF-1α/VEGF pathway exhibited up-regulated expression, promoting ovarian angiogenesis in POI rats, and enhancing follicular development and granulosa cell survival, thereby restoring fertility in rats. CONCLUSIONS: DHEA can potentially restore ovarian function in rats with cyclophosphamide-induced POI by up-regulating HIF-1α/VEGF signalling, which promotes the growth of blood vessels in the ovaries.

Melatonin-pretreated human umbilical cord mesenchymal stem cells improved endometrium regeneration and fertility recovery through macrophage immunomodulation in rats with intrauterine adhesions†.

Intrauterine adhesions (IUA) are a common gynecological problem. Stem cell therapy has been widely used in the treatment of IUA. However, due to the complex and harsh microenvironment of the uterine cavity, the effectiveness of such therapy is greatly inhibited. This study aimed to investigate whether melatonin pretreatment enhances the efficacy of human umbilical cord mesenchymal stem cells (HucMSCs) in IUA treatment in rats. First, we explored the effect of melatonin on the biological activity of HucMSCs in vitro through a macrophage co-culture system, Cell Counting Kit 8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), flow cytometry, immunofluorescence staining, and qRT-PCR. Subsequently, we established the IUA rat model and tracked the distribution of HucMSCs in this model. In addition, we observed the number of M1 and M2 macrophages through immunofluorescence staining and detected the levels of inflammatory cytokines. Four weeks after cell transplantation, HE, Masson, and immunohistochemical staining were performed. In vitro experiments showed that melatonin pretreatment of HucMSCs promoted proliferation, reduced apoptosis, up-regulated the stemness gene, and regulated macrophage polarization. In vivo, melatonin pretreatment caused more HucMSCs to remain in the uterine cavity. Melatonin-pretreated HucMSCs recruited more macrophages, regulated macrophage polarization, and reduced inflammation. Melatonin-pretreated HucMSCs relieved fibrosis, increased endometrium thickness, and up-regulated CD34, vimentin, proliferating cell nuclear antigen (PCNA), and alpha small muscle antigen (α-SMA) expression. Fertility tests showed that melatonin-pretreated HucMSCs increased the number of embryos. In summary, pretreatment with melatonin was beneficial for HucMSC treatment because it enhanced the cell's ability to recruit macrophages and regulate macrophage polarization, which led to the regeneration of the endometrium and improved pregnancy outcomes.

Aquaporin 9 causes recurrent spontaneous abortion by inhibiting trophoblast cell epithelial-mesenchymal transformation and invasion through the PI3K/AKT pathway†.

BACKGROUND: Invasion of the endometrium by trophoblast cells is a key event during pregnancy, although the underlying mechanism remains unclear. Aquaporin 9 (AQP 9) is expressed in many eukaryotes and is associated with cell invasion. The objective of this study was to evaluate the significance of AQP9 in recurrent spontaneous abortion. METHODS: We screened the GSE22490 dataset and further differentiated aquaporin 9 expression in villi. AQP9 was evaluated as one of the key factors in abortion by injecting AQP9 overexpressed plasmid into the uterus of CD1 mice. Trophoblast cells were transfected with AQP9-overexpressing plasmid or siAQP9 to measure cell proliferation, migration, invasion, and apoptosis. Western blot was used to measure changes in the expression of invasion, epithelial-mesenchymal transformation process, and PI3K/AKT pathway. Finally, the role of AQP9 in PI3K/AKT signaling pathway was determined using the PI3K/AKT inhibitor, LY294002, and activator, 740Y-P. RESULTS: AQP9 is highly expressed in recurrent spontaneous abortion villus. Intrauterine injections of AQP9-overexpressing plasmid into CD1 mice resulted in atrophy and blackness of the gestational sac and increased the absorption rate, it is the causative factor of abortion. AQP9 upregulation inhibited the proliferation, invasion, migration, and epithelial-mesenchymal transformation process in vitro of trophoblast cells and increased cell apoptosis. The opposite result was observed after silencing AQP9. AQP9 overexpression also inhibited the PI3K/AKT pathway. LY294002 and 740Y-P partially recovered AQP9-induced trophoblast invasion and migration via the PI3K/AKT pathway. CONCLUSIONS: AQP9 reduces the invasive ability of trophoblast cells by regulating PI3K/AKT signaling pathway, participating in recurrent spontaneous abortion.

Progranulin promotes proliferation, migration and invasion via the PI3K/Akt signalling pathway in a model of endometriosis.

RESEARCH QUESTION: What are the levels of progranulin (PGRN) expression in primary endometrial stromal cells (ESC) and endometrial tissue in patients with endometriosis (EMS)? What is the role and mechanism of action of PGRN in EMS? DESIGN: Endometrial tissue was collected from 30 patients, 15 with EMS (EMS group) and 15 without EMS (non-EMS group). PGRN expression in endometrial tissue and ESC was analysed by immunohistochemistry, immunofluorescence, western blotting and quantitative reverse transcription polymerase chain reaction. PGRN overexpression and silencing ESC were established with lentivirus to detect the effect on proliferation, invasion and migration. The relationship between PGRN and the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) signalling pathway was verified by western blotting. A rescue assay was performed with PI3K inhibitor treatment. RESULTS: The PGRN expression was significantly higher in EMS samples. PGRN up-regulation promoted proliferation (P = 0.007), migration (P = 0.002) and invasion (P < 0.001) of eutopic endometrial stromal cells (EUESC). The ratio of p-AKT/AKT was higher in the overexpression PGRN (ovPGRN) group than in the overexpression-NC (ovNC) group (P = 0.004). Silencing PGRN produced the opposite results, and LY2940002 addition reversed the effect of PGRN up-regulation on the proliferation, invasion and migration of EUESC. CONCLUSIONS: PGRN might promote the proliferation, invasion and migration of EUESC via the PI3K/Akt signalling pathway. These preliminary in-vitro findings may present a new perspective and inspire further study of the mechanism of EMS.

Combined oral antibiotics and intrauterine perfusion can improve in vitro fertilization and embryo transfer pregnancy outcomes in patients with chronic endometritis and repeated embryo implantation failure.

BACKGROUND: The aim of this retrospective study was to investigate whether oral antibiotics (doxycycline and metronidazole) combined with intrauterine perfusion (gentamicin and dexamethasone) are beneficial for patients with repeated implantation failure (RIF) and chronic endometritis (CE) to improve clinical pregnancy outcomes. METHODS: Patients with RIF and CE were diagnosed using hysteroscopy and histology together. A total of 42 patients were enrolled in the study. All patients received oral antibiotics (doxycycline combined with metronidazole) and 22 patients underwent intrauterine perfusion (gentamicin combined with dexamethasone) immediately after the end of oral antibiotic therapy. Pregnancy outcomes were evaluated during the first in vitro fertilization (IVF) and embryo transfer (ET) cycle. RESULTS: For the first D3 ET after treatment with oral antibiotics (doxycycline and metronidazole) combined with intrauterine perfusion (gentamicin and dexamethasone), higher embryo implantation rate (30.95% vs. 26.67%, P = 0.0308), clinical pregnancy rate (30% vs. 50%, P < 0.001), live birth rate (33.33% vs. 45.45%, P < 0.0001). No fetal malformations or ectopic pregnancies were observed. CONCLUSION: We report oral antibiotics (doxycycline and metronidazole) combined with intrauterine perfusion (gentamicin and dexamethasone) as a novel treatment for CE to improve the outcomes of successful pregnancy compared with those of oral antibiotics alone.

The diagnostic value of serum YKL-40 in endometrial cancer: a meta-analysis.

BACKGROUND: Serum YKL-40 is a promising non-invasive biomarker for the early diagnosis of endometrial cancer (EC), but its value is disputed. OBJECTIVE: To investigate the serum YKL-40 in the early diagnostic value of EC. METHODS: Databases were systematically searched again before April 2021 and 14 studies were finally included in this meta-analysis. Pooled sensitivity, specificity, negative likelihood ratio (NLR), positive likelihood ratio (PLR), diagnostic odds ratio (DOR), and summary receiver operating characteristic (SROC) curve analyses were assessed. This meta-analysis investigated the source of heterogeneity using sensitivity analysis, subgroup analysis, and meta-regression. RESULTS: Databases were systematically searched again before April 2021 and 14 studies were finally included in this meta-analysis. First, the SROC curve presented an area under the curve (AUC) of 0.853 (SE = 0.0213) for YKL-40 alone and an AUC of 0.946 (SE = 0.0268) for YKL-40 combined with other biomarkers. Second, diagnostic types might be related to the diagnostic accuracy and is a significant source of heterogeneity (P = 0.035). CONCLUSION: Serum YKL-40 helped diagnose EC, and its combination with other biomarkers was better than itself alone.

Oral immunosuppressants improve pregnancy outcomes in women with idiopathic recurrent miscarriage: A meta-analysis.

WHAT IS KNOWN AND OBJECTIVE: Reports said immunotherapy is effective for the treatment of idiopathic recurrent miscarriage (RM). Immunotherapy is invasive, and lymphocyte therapy carries some risk of infection. Oral immunosuppressants have the advantages of simple administration and convenience; however, there is no statistical analysis of whether they can improve pregnancy outcomes in patients with idiopathic RM. METHODS: Six databases were searched for studies on oral immunosuppressants and RM; 374 articles were identified. There were two oral immunosuppressants, cyclosporine A and prednisone; two studies were on cyclosporine A and three studies were on prednisone for RM. RESULTS AND DISCUSSION: In total, 554 RM patients were included in this analysis, including 357 patients who received oral immunosuppressive agents and 197 patients who received basic treatment, placebo, or no treatment. Oral administration of cyclosporine A or prednisolone increases live birth rate (OR = 3.6, 95% CI: 2.1-6.15, p < 0.00001) and ongoing pregnancy rate (OR = 8.82, 95% CI: 2.91-26.75, p = 0.0001) in patients with idiopathic RM. Drug use reduced miscarriage rate (OR = 0.21, 95% CI: 0.08-0.52, p = 0.0007); however, there was significant heterogeneity (I2  = 73%) and a moderate-to-severe risk of bias. There was no effect on premature birth rate (OR = 2.26, 95% CI: 0.96-5.31, p = 0.06). This meta-analysis cannot provide a reference for the duration of medication treatment because the selected studies had inconsistent durations. WHAT IS NEW AND CONCLUSION: We did a statistical analysis and found that oral immunosuppressants (including cyclosporine A or prednisolone) can improve pregnancy outcomes in patients with idiopathic RM, increase live birth rate and ongoing pregnancy rate, and reduce miscarriage rate.

Injectable collagen hydrogel combines human umbilical cord mesenchymal stem cells to promote endometrial regeneration in rats with thin endometrium.

The regeneration of thin endometrium still remains as a great challenge in the field of reproductive medicine. Stem cells-based therapy has been considered as a promising strategy for the restoration of thin endometrium. However, the low transplantation and retention rate of stem cells and loss of stemness due to in vitro expansion limits the therapeutic efficacy. In our study, we combined collagen hydrogel and human umbilical cord mesenchymal stem cells (uMSCs) for improving the regeneration of thin endometrium, by using the potent pluripotency and low immunogenicity of uMSCs and collagen hydrogel that promotes the anchorage and proliferation of stem cells. Results showed that collagen hydrogel has favorable biocompatibility and the capacity to enhance the cell viability and expression of stemness-associated genes (including organic cation/carnitine transporter4 (Oct-4), Nanog homeobox (Nanog) and SRY-box transcription factor 2 (SOX2)) of uMSCs. The combination of collagen hydrogel and uMSCs prolonged the retention time of the constructs in the uterine cavity and improved endometrial thickness compared with uMSCs alone, leading to increase the fertility of the rats with thin endometrium. These highlighted therapeutic prospects of collagen hydrogel combined with uMSCs for the minimally invasive therapy of thin endometrium in the clinic.

Fever glove hand-shake method safe blood collection from children's fingertips in COVID-19 fever clinic.

BACKGROUND: During the coronavirus disease 2019 (COVID-19) epidemic, the fever clinic is an important link for screening and diagnosing whether a patient is infected with the novel coronavirus. Blood collection from children's fingertips is a commonly used detection method; however, in children, the blood collection process may cause discomfort and resistance. To address this problem, the use of heating gloves combined with hand swinging can be considered for fingertip blood collection in children. AIM: To explore the application of fever gloves with the handshaking method for fingertip blood collection from children in fever clinics during the COVID-19 epidemic. METHODS: A total of 100 children were selected for fingertip blood collection at the fever clinic of our hospital from June 2022 to June 2023 and were divided into two groups using a randomized numerical table method, with 50 cases in each group, including the control and observation groups. The patients in the control group followed the doctor's instructions to cooperate with the routine fingertip blood collection method, and the patients in the observation group followed the doctor's instructions to cooperate with the static fever gloves with the shaking hands method of children's fingertip blood collection. The level of the six blood routine and collection indexes, and the satisfaction of the examination of the patients in the peripheral blood group and the fever gloves with the shaking hands method of the children's fingertip blood collection group were compared. RESULTS: The red and white blood cell count, hemoglobin, and red blood cell pressure volume in the observation group were higher than those in the control group (P < 0.05); the platelet count in the control group was lower than that in the observation group (P < 0.05); the number of times of squeezing the fingertip, the average time of blood collection, and the score of puncture pain in the observation group were significantly better than those in the control group (P < 0.05); and satisfaction with the routine blood examination in the observation group was greater than that in the control group. CONCLUSION: The application value of the fever gloves with shaking hands method for children's fingertip blood collection was better, the accuracy of examination indexes was higher, and patient satisfaction with the examination was greater.

Construction and In Vitro Digestibility of Recrystallized Starch Encapsulated in Calcium Alginate Beads.

In order to reduce the digestion rate of starch in human body and improve the content of slowly digestible starch (SDS) and resistant starch (RS), millimeter calcium alginate beads encapsulated with different proportions of recrystallized starch were constructed in this study. First, we prepared recrystallized starch (RS3) by debranching waxy corn starch and retrogradation, and then encapsulated RS3 in calcium alginate beads by the ionic gel method. The microstructure of the beads was observed by scanning electron microscope, and the gel texture properties, swelling properties, and in vitro digestibility of the beads were studied. The results showed that the beads after cooking still maintained high hardness and chewiness, and the swelling power and solubility of the beads were lower than that of native starch. Compared with native starch, the content of rapidly digestible starch (RDS) in beads decreased, while the content of SDS and RS increased. The sample with the highest content of RS is RS31@Alginate1, whose content of RS is 70.10%, 52.11 times higher than that of waxy corn starch and 1.75 times higher than that of RS3. RS3 encapsulated in calcium alginate beads has a good encapsulation effect, and the content of SDS and RS is greatly increased. This study has important implications for reducing the digestion rate of starch and regulating the health of people with diabetes and obesity.

Activated Human Umbilical Cord Blood Platelet-Rich Plasma Enhances the Beneficial Effects of Human Umbilical Cord Mesenchymal Stem Cells in Chemotherapy-Induced POF Rats.

Saving the ovarian function of premature ovarian failure (POF) patients undergoing chemotherapy is an important problem in the field of reproductive medicine. At present, human umbilical cord mesenchymal stem cells (HucMSCs) have been used in the treatment of POF, but the effect is still not optimal. The purpose of this study was to determine whether human umbilical cord blood platelet-rich plasma (ucPRP) enhances the beneficial effects of HucMSCs in the treatment of POF. First, we observed the effects of changes in the biological activity of ucPRP on HucMSCs in vitro. Subsequently, we tracked the distribution and function of the HucMSCs in POF rats, and the rats' estrus cycle and serum sex hormones, follicular development, ovarian angiogenesis, ovarian granulosa cell proliferation, and apoptosis were assessed. The results of the study showed that the addition of ucPRP in vitro accelerates proliferation and reduces apoptosis of the HucMSCs while upregulating the stemness gene of the HucMSCs. The combined transplantation of HucMSCs and ucPRP resulted in more stem cells being retained in the ovaries of POF rats, the estrus cycle of the POF rats being restored, the levels of serum E2, AMH, and FSH improving, and damaged follicles beginning to grow. Finally, we confirmed that the potential mechanism of the combination of HucMSCs and ucPRP to rescue the ovarian function of POF rats is to promote ovarian angiogenesis and to promote the proliferation and reduce the apoptosis of ovarian granulosa cells. The upregulation of AMH and FHSR expression and the downregulation of caspase-3 expression in granulosa cells are potential mechanisms for the recovery of ovarian function. Our research results suggest that the combined application of HucMSCs and ucPRP is a safe and efficient transplantation program for the treatment of POF, thus providing a reliable experimental basis for the clinical application of stem cell therapy in POF.