Advances in mesenchymal stem cells therapy for tendinopathies.

Tendinopathies are chronic diseases of an unknown etiology and associated with inflammation. Mesenchymal stem cells (MSCs) have emerged as a viable therapeutic option to combat the pathological progression of tendinopathies, not only because of their potential for multidirectional differentiation and self-renewal, but also their excellent immunomodulatory properties. The immunomodulatory effects of MSCs are increasingly being recognized as playing a crucial role in the treatment of tendinopathies, with MSCs being pivotal in regulating the inflammatory microenvironment by modulating the immune response, ultimately contributing to improved tissue repair. This review will discuss the current knowledge regarding the application of MSCs in tendinopathy treatments through the modulation of the immune response.

Letrozole + Ziwu Liuzhu Transcutaneous Electrical Acupoint Stimulation Improves Ovulation-Induced Pregnancy Rate in Obese Women with Polycystic Ovary Syndrome: A Prospective Cohort Study.

Objective: This study investigated the effect of letrozole with/without meridian-infusion percutaneous electrical stimulation on the rates of ovulation-induced pregnancy in patients with obese polycystic ovary syndrome (obPCOS). Materials and Methods: Patients with obPCOS, ages 20-40, each with a body mass index (BMI) ≥24 kg/m2, and/or waist circumference ≥80 cm, and at least 1 side tubal patency were enrolled at the Guangdong Women and Children Hospital, Guangzhou, China. They were divided into 2 groups: ZLT [Ziwu Liuzhu + transcutaneous electrical acupoint stimulation] and control. Baseline conditions and pregnancy status were collected for all patients. Multivariate Cox regression analysis and sensitivity analysis of propensity score matching (PSM) were performed for the groups after multiple interpolations. Results: From July 2021 to September 2022, 345 patients with obPCOS were recruited: 53 cases/69 cycles in the ZLT group and 292 cases/396 cycles in the control group. The 2 sets of baselines were flush. The anovulatory cycle rates were: ZLT, 2.89% (2/69); and control, 1.77% (7/396); P > 0.05. Multifollicle growth-cycle rates were: ZLT, 0% (0/69); and control, 0.76% (3/396); P > 0.05. Multivariate COX regression analysis showed adjusted hazard ratio (aHR) 95% confidence interval (CI): 2.11 (1.19, 3.73); P = 0.011. Multivariate Cox regression analysis with multiple imputation showed aHR 95% CI: 2.11 (1.19, 3.73); P = 0.013. In the overweight group (24-28 kg/m2), the pregnancy rate of the control and ZLT groups were 20.2% and 32.3%, respectively, aHR 95% CI: 1.76 (0.87,3.55); P = 0.113. In the obese cohort (≥ 28 kg/m2), the control and ZLT groups, pregnancy rates were 10.7% and 27.3%, respectively, aHR 95% CI: 3.46 (1.21, 9.92); P = 0.021; (Pfor interaction = 0.369). The caliper value was set as 0.2 for BMI and antral-follicle count, and PSM was performed at 1:1, aHR 95%CI: 2.45 (1.01, 5.96); P = 0.048. Conclusions: Letrazole + ZLT had a positive effect on ovulation-induced pregnancy rates in patients with obPCOS.

In vitro maturation of human oocytes maintaining good development potential for rescue intracytoplasmic sperm injection with fresh sperm.

BACKGROUND: The outcomes of the use of commercial in vitro maturation (IVM) medium to culture immature oocytes obtained from conventional ovulation induction, followed by rescue intracytoplasmic sperm injection (RICSI), are not ideal. It is thus difficult to widely adopt this approach in clinical practice. Therefore, it is necessary to explore methods for improving the clinical outcome of IVM. AIM: To study the effect of sperm on the developmental potential of in vitro-matured oocytes in conventional culture. METHODS: This was a retrospective study of patients whose immature oocytes were harvested from conventional oocyte stimulation cycles and underwent ICSI at our hospital between June 2018 and August 2020. RICSI was performed using sperm collected on the day of oocyte harvest (old) and sperm collected on the day of RICSI (fresh) and oocytes matured in vitro after 24 h of culture in conventional medium. The rates of in vitro oocyte maturation, normal fertilization, normal cleavage, day-3 top-quality embryos, and useful blastocyst formation were compared between the two groups. RESULTS: In total, 102 germinal vesicle (GV)-stage immature oocytes were cultured in the old sperm group. In the fresh sperm group, 122 GV-stage immature oocytes were collected and cultured in vitro for 24 h. There were no significant differences in the general conditions of males and females between the two groups (P > 0.05). The oocyte maturation, normal fertilization, and normal cleavage rates of the old and fresh groups were 51.0% vs 55.7%, 61.5% vs 64.7%, and 93.8% vs 93.2%, respectively. None of the rates differed significantly (P > 0.05) between the two groups. However, the day-3 top-quality embryo and useful blastocyst rates of the old and fresh sperm groups were 16.6% vs 63.4%; 6.67% vs 34.6%, respectively. The day-3 top-quality embryos and useful blastocyst rates of the old sperm group were significantly lower than those of the fresh group (P < 0.05). CONCLUSION: In vitro maturation with conventional culture medium combined with the use of fresh sperm collected on the day of RICSI is an easy-to-implement strategy for patients whose oocytes are completely or mostly immature.

Association between baseline LH/FSH and live-birth rate after fresh-embryo transfer in polycystic ovary syndrome women.

This study aimed to retrospectively analyse the effect of the baseline luteinising hormone/follicle-stimulating hormone ratio (bLH/FSH) on the live-birth rate per fresh-embryo transfer cycle (LBR/ET) in infertile women with polycystic ovary syndrome (PCOS) who received a fresh-embryo transfer. A total of 424 patients with PCOS who underwent the first cycle of in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI) fresh-embryo transfer at our hospital was enrolled. Univariate and multivariate logistic regression analyses, along with curve fitting and a threshold effect analysis, were performed. Baseline LH/FSH levels were a significant (P < 0.05) independent risk factor affecting live birth. In the first IVF/ICSI antagonist treatment cycles, LBR/ET after fresh-embryo transfer was relatively flat, until bLH/FSH was 1.0; thereafter, it started to decrease by 17% for every 0.1-unit bLH/FSH increase. Considering the decline in LBR/ET, it is recommended that PCOS women with bLH/FSH > 1.0 carefully consider fresh-embryo transfer during their first IVF/ICSI.

The Clinical Efficacy of Three Different Follicle-Stimulating Hormones for Follicle Growth and Development in Long-Protocol Controlled Ovarian Hyperstimulation Treatment.

OBJECTIVE: To compare the use and clinical efficacy of three different follicle-stimulating hormones (FSHs) for follicle growth and development in long-protocol controlled ovarian hyperstimulation (COH). METHODS: A total of 540 gonadotropin-releasing hormone (GnRH) agonists' long protocol treatment cycles at our hospital between January 2015 and May 2020 and met the inclusion criteria were retrospectively analyzed. The cycles were divided into three groups based on their indexes (groups A, B, and C). Each of the groups received a different type of FSH during treatment. A cross-group comparison was then undertaken to evaluate the growth and development of the three largest follicles and the patients' pregnancy-related indexes between the normal-response and high-response populations. RESULTS: In the normal-response populations, the number of high-quality embryos obtained in groups A and B was significantly higher than in group C, and the FSH dosage was significantly lower than in group C (P < 0.05). There were more follicles with a diameter of 16-18 mm found in group A than in group C on the day of hCG injection (hCG day) (P < 0.05), but there were no significant differences in the groups in other indicators. In the high-response populations, the number of oocytes retrieved and high-quality embryos obtained in group A were significantly higher than in group C (P < 0.05), and the total dosage and duration of FSH stimulation in group C were significantly higher than groups A and B (P < 0.05). CONCLUSION: Three different types of FSH led to comparable growth rates of the three largest follicles and clinical pregnancy rates per fresh cycle in long-protocol COH treatment.

Comparison of vaginal progesterone gel combined with oral dydrogesterone versus intramuscular progesterone for luteal support in hormone replacement therapy-frozen embryo transfer cycle.

BACKGROUND: It remains under subject of debate regarding the optimal route of luteal support for hormone replacement therapy- frozen embryo transfer (HRT-FET) cycles. We compared efficacy of vaginal progesterone gel combined with oral dydrogesterone and intramuscular progesterone for HRT-FET lutein support. METHODS: This is a retrospective observational study. After matching for propensity score of getting vaginal + oral treatment, a total of 208 FET cycles in the vaginal progesterone combined with oral dydrogesterone and 624 cycles in the intramuscular progesterone group were enrolled. Pregnancy outcomes and neonatal outcomes including chemical pregnancy rate, clinical pregnancy rate, implantation rate, spontaneous abortion rate, live birth rate, gestational weeks, pre-term delivery, birth weight, and congenital anomalies rate were compared. RESULTS: No significant differences were observed in patient characteristics such as age, duration of infertility, type of infertility, or hormone level after matching. Chemical pregnancy rate (68.3 % versus 70.5 %), clinical pregnancy rate (64.9 % versus 64.4 %), implantation rate (52.3 % versus 50.2 %), spontaneous abortion rate (21.5 % versus 18.4 %), and live birth rate (49.0 % versus 51.3 %) were similar in both group without statistically significant difference. No significant differences in neonatal outcomes were observed between the two groups. CONCLUSION: We observed similar pregnancy outcomes in both vaginal progesterone gel combined with oral dydrogesterone and intramuscular progesterone protocol. Vaginal progesterone gel combined with oral dydrogesterone can be substituted for intramuscular progesterone given that vaginal plus oral use has good safety and is more convenient and may be associated with less side effect caused by intramuscular injection.

The impacts of the number of prefreeze and postthaw blastomeres on embryo implantation potential: A systematic analysis.

To systematically analyze the potential of embryo implantation through comparison between the number of surviving blastomeres, the growth, and implantation rate.Retrospective analysis on implantation rate and the growth of prefreeze-postthaw embryos with different blastomeres in 1487 frozen embryo transfer cycles.In groups of postthaw embryos without damage, implantation rate and the average number of blastomere growth increased significantly with increasing number of blastomeres. The implantation rate and the number of blastomeres of embryos with 8-8c (the number of blastomeres in prefreeze embryo-the number of blastomeres in postthaw embryo) continued to grow at a significantly higher rate than that of 5-5c and 6-6c (P < .05). In groups of embryos with the same number of blastomeres before freezing and with partial damage after resuscitation, the implantation rates were lower and the average numbers of blastomere growth reduced as the number of damaged blastomeres increased. For embryos with good quality before freezing, 1 to 3 damaged blastomeres in postthawed embryos did not affect the development and implantation rate. Both implantation rate and growth rate of embryos with 8-6c were significantly higher than those of embryos with 6-6c (P < .05).The number of surviving blastomeres and growth in frozen-thawed embryos could be important index to predict embryo development potential and clinical outcome of implantation. For embryos with good quality, a small amount of damaged blastomeres would not weaken embryo development potential and implantation rate after being thawed.

Synaptotagmin 1 regulates cortical granule exocytosis during mouse oocyte activation.

Synaptotagmin 1 (Syt1) is an abundant and important presynaptic vesicle protein that binds Ca2+ for the regulation of synaptic vesicle exocytosis. Our previous study reported its localization and function on spindle assembly in mouse oocyte meiotic maturation. The present study was designed to investigate the function of Syt1 during mouse oocyte activation and subsequent cortical granule exocytosis (CGE) using confocal microscopy, morpholinol-based knockdown and time-lapse live cell imaging. By employing live cell imaging, we first studied the dynamic process of CGE and calculated the time interval between [Ca2+]i rise and CGE after oocyte activation. We further showed that Syt1 was co-localized to cortical granules (CGs) at the oocyte cortex. After oocyte activation with SrCl2, the Syt1 distribution pattern was altered significantly, similar to the changes seen for the CGs. Knockdown of Syt1 inhibited [Ca2+]i oscillations, disrupted the F-actin distribution pattern and delayed the time of cortical reaction. In summary, as a synaptic vesicle protein and calcium sensor for exocytosis, Syt1 acts as an essential regulator in mouse oocyte activation events including the generation of Ca2+ signals and CGE.

[Determination of Desloratadine and Its Metabolite 3-OH Desloratadine in Human Plasma by LC-MS/MS].

OBJECTIVE: To develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of desloratadine and its metabolite 3-OH desloratadine in human plasma. METHODS: 24 healthy male volunteers received a single oral dose of 5 mg desloratadine tablets in a randomized crossover bioequivalence study with two preparations of tablets. Serial plasma samples were taken and analyzed by the LC-MS/MS method. The pharmacokinetic parameters of the two preparations were calculated and compared statistically to evaluate their bioequivalence using Winnonlin 6. 3. RESULTS: The calibration curves of desloratadine and 3-OH desloratadine were both linear over the concentration range of 0. 050-6. 0 ng/mL, with intra-batch and inter-batch relative standard deviations less than 15%. The 90% confidence intervals (CIs) of peak concentration (Cmax) area under the curve (AUC)0t and AUC0-∞ of desloratadine and 3-OH desloratadine all resided within the bioequivalence limit 80%-125%. No significant difference in peak time (Tmax) was demonstrated between the two preparations. CONCLUSION: The LC-MS/MS method can be used for simultaneous determination of desloratadine and 3-OH desloratadine in human plasma, which has been successfully applied-to a bioequivalence study.

[Co-culture of human extravillous cytotrophoblasts and decidual stromal cells in vitro].

OBJECTIVE: To develop a convenient method for isolation and purification of human extravillous cytotrophoblasts (EVCTs) and decidual stromal cells (DSCs) and establish a co-culture system. METHODS: The DSCs were digested with trypsin and purified by Percoll gradient. The EVCTs were digested with trypsin and purified by BSA gradient. Immunohischemistry and immunofluorescent study are performed to characterize these isolated cells. The EVCTs and DSCs were placed in Matrigel-coated Transwell upper and lower chamber, respectively, to study the invasive ability of the EVCTs. RESULTS: Immunohischemistry revealed that the purity of EVCTs and DSC exceeded 95%. Cultured EVCTs retained their capacity to invade Matrigel-coated Transwell filters with the invasion index of 3.22-/+0.04. CONCLUSION: This co-culture model established by isolating highly purified EVCTs and DSCs in vitro can be useful for studying the trophoblast invasion mechanisms.

[Synthesis and luminescence properties of ternary complexes of europium with aromatic carboxylic acid and acrylonitrile].

Five ternary complexes were synthesized from europium with aromatic carboxylic acid (p-methylbenzoic acid, methoxybenzoic acid, m-chlorobenzoic acid and benzoic acid, p-hydroxylbenzoic acid) and acrylonitrile, and characterized by means of elemental analysis, thermal analysis, FTIR spectra and UV spectra. The fluorescence spectra show that five ternary complexes have good luminescence properties, and the sequence of the ability of the aromatic carboxylic acids to transfer light energy to europium ion is as follows: p-methylbenzoic acid>benzoic acid>m-chlorobenzoic acid>p-hydroxylbenzoic acid>methoxybenzoic acid. Meanwhile, the ternary europium complexes containing a reactive ligand acrylonitrile will possibly have a potential application to the fabrication of bonding-type europium polymer luminescent materials.

[Effects of mitogen-activated protein kinase on phorbol 12-myristate 13-acetate-induced matrix metalloproteinase-9 gene expression in cytotrophoblastic cells].

OBJECTIVE: To explore the regulatory mechanism of matrix metalloproteinase-9 (MMP-9) gene expression induced by phorbol 12-myristate 13-acetate (PMA) in cytotrophoblastic cells. METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to determine the kinase activity, and MMP-9 gene expression was detected by semi-quantitative reverse transcription (RT)-PCR in the cytotrophoblastic cells. RESULTS: Cytotrophoblastic cells treated with PMA showed markedly increased MMP-9 mRNA level. PMA treatment caused an increase in extracellular signal-regulated kinase (ERK), c-jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38 MAPK) activities. Both SB203580 (specific inhibitor of the p38 MAPK) and PD98059 (specific inhibitor of the ERK), when preincubated for 30 min with the cytotrophoblastic cells, substantially reduced MMP-9 mRNA accumulation in PMA-primed cells. However, neither SB203580 nor PD98059 used alone, or their combination, was able to completely inhibit MMP-9 mRNA expression. CONCLUSION: Both p38 MAPK and ERK pathways are involved in the regulation of MMP-9 gene expression in the cytotrophoblastic cells induced by PMA, and both signaling pathways are indispensable for full activation of MMP-9 gene expression.

[Role of p38 pathway in PMA-induced in vitro invasion of JAR human choriocarcinoma cell line].

OBJECTIVE: To investigate the role of p38 mitogen-activated protein kinase (MAPK) signal transduction pathways in regulating the in vitro invasion of JAR human choriocarcinoma cells induced by phorbol 12-myristate 13-acetate (PMA). METHODS: ELISA was used to detect the kinase activity of the JAR cells in response to PMA stimulation, and the in vitro invasion capabilities of the stimulated cells were observed using transwell assay. Changes in the proliferation and activity of the JAR cells cultured in vitro following PMA treatment were also observed by MTT assay. RESULTS: p38 MAPK was activated dose-dependently in JAR cells upon the stimulation by PMA, which significantly enhanced the in vitro invasion of the JAR cells, while treatment of the cells with SB203580, a specific inhibitor of p38 MAPK, inhibited the invasion of the cells. The growth of the cells, as observed from the growth curves, was not affected by the treatment of PMA and/or SB203580. CONCLUSION: Activation of p38 MAPK signal transduction pathway may enhance the invasion capability of JAR cells, and p38 MAPK inhibition may therefore yield new possibility to control the invasion of choriocarcinoma.

[Role of phorbol 12-myristate 13-acetate on cytokine expression in JAR trophoblast cell line].

OBJECTIVE: To investigate whether phorbol 12-myristate 13-acetate (PMA) modifies the invasive ability of trophoblast cells by regulating their cytokine productions. METHODS: Reverse transcriptase-polymerase chain reaction was used to examine the effect of PMA on the expression of cytokines which regulated the invasive ability of trophoblast cells. RESULTS: Prior to PMA treatment, expressions of the cytokins including hepatocyte growth factor (HGF), interleukin (IL)-1beta, insulin-like growth factor (IGF)-II, transforming growth factor (TGF)- beta and vascular endothelial growth factor (VEGF) were all detected in JAR cells, only with the exception of IGF-I. After incubation with 100 nmol/L PMA for 24 h, the cells showed strong expression of IL-1beta, HGF and IGF-II, with reduced expression of TGF-beta2 and TGF-beta3. CONCLUSION: By regulating the autocrine of these cytokines, PMA exercises its effect to enhance the invasive ability of trophoblast or choriocarcinoma cells.

Effect of conditioned media from decidual cell culture on the expression of genes regulating the invasion of trophoblastic cells.

OBJECTIVE: To investigate the effect of the conditioned media from decidual cell cultures (DCM) on the expression of genes regulating the invasion of trophoblastic cells. METHODS: In vitro culture of decidual cells obtained from healthy women of both early (within the first trimester) and full-term pregnancy respectively was performed to prepare conditioned media of decidual cells (DCM). Trophoblastic cells were also obtained in these subjects and treated with DCM for 24 h in in vitro culture, to observe the effect of DCM, with the help of semi-quantitative reverse transcriptase-PCR, on the expression of genes in these cells that regulate their invasion. RESULTS: Expression of matrix metallsoproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinases (TIMP)-1 and urokinase-type plasminogen activator (u-PA), other than that of TIMP-2 and plasminogen activator inhibitor type (PAI)-1, was observed in normal trophoblastic cells in in vitro culture. DCM derived from wemon of early and full-term pregnancy down-regulated the expression of MMP-2, MMP-9, u-PA while up-regulated the expression of TIMP-1, PAI-1. CONCLUSION: DCM may exhibit its anti-invasive activity by regulating the expression of genes involved in the regulation of trophoblastic cell invasion, such as MMP-2, MMP-9, TIMP-1, u-PA and PAI-1.