OBJECTIVE: To observe the effect of "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique on the reproductive function of the mice with poor ovarian response (POR) and explore the molecular mechanism of acupuncture on early embryos after in vitro fertilization-embryo transfer (IVF-ET) in POR mice. METHODS: Of 70 female C57BL/6 mice, 60 mice with regular estrous cycle were screened and 30 of them were randomly divided into a blank group, a model group, and an acupuncture group, 10 mice in each one. Mice in the model and acupuncture groups were administered with tripterygium glycosides suspension (50 mg·kg^-1·d^-1) via gavage for 2 weeks to prepare POR models; while in the blank group, the mice received an equal volume of 0.9% sodium chloride solution via gavage. After successful modeling, mice in the acupuncture group underwent "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique, once daily, for 20 min each time and lasting 2 consecutive weeks. After intervention completion, subsequently, all the three groups underwent ovulation induction, orbital blood collection, and ovary extraction and fresh denuded oocyte collection. Denuded oocytes, after incubated, together with the sperms from 15 male C57BL/6 mice, were transferred into the oviducts of 30 donor pseudopregnant C57BL/6 female mice via IVF-ET; and the embryonic tissue was collected on day 7 of implantation. After successful modeling and intervention completion, the general conditions of mice in each group were observed, and the estrous cycle disorder rate was compared among the groups. After intervention completion, the numbers of oocyte, ovarian wet weight and ovarian index were recorded in each group; the levels of anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) in the serum were measured using ELISA; and ovarian morphology and mitochondrial ultrastructure were examined using HE staining and transmission electron microscopy. The transcriptome sequencing technology was employed to identify differentially expressed genes (DEGs) in early embryos of each group, followed by bioinformatics analysis; and the reversed DEGs with significant difference were verified using quantitative real-time PCR (qRT-PCR). RESULTS: After intervention, compared with the blank group, the mice in the model group presented poor spirits and declined water and food intake, reduced activity, the higher rate of estrous cycle disorder (P< 0.01); the decrease of the numbers of oocyte, ovarian wet weight, and ovarian index (P<0.01), reduced AMH and E2 in the serum (P<0.01), elevated FSH and LH in the serum (P<0.01); more atresia follicles in ovarian tissue, disorganized granulosa cells, reduced the numbers of mitochondria, vacuoles appeared and cristae breakage. When compared with the model group, in the acupuncture group, the spirits of mice, food intake and activity were improved, the rate of estrous cycle disorder was reduced (P<0.01); the numbers of oocyte, ovarian wet weight, and ovarian index were higher (P<0.01, P<0.05), the levels of AMH and E2 in the serum increased (P<0.01), the levels of FSH and LH in the serum decreased (P<0.01); the atresia follicles were reduced, the numbers of follicle increased at all levels, the numbers of mitochondria got higher, vacuoles and cristae breakage were declined. In comparison between the blank group and the model group, and between the model group and the acupuncture group, a total of 258 intersecting DEGs were identified, and 194 DEGs of them exhibited a trend of reversion before and after intervention. The top 5 reversed DEGs with significant difference included dexamethasone-induced Ras-related protein 1 (Rasd1), gene regulated by estrogen in breast cancer 1 (Greb1), leucine-rich repeat-containing G protein-coupled receptor 6 (Lgr6), Fraser syndrome 1 (Fras1), and apolipoprotein D (Apod). Compared with the blank group, the mRNA expression of Rasd1, Greb1, Lgr6, Fras1 and Apod in embryonic tissues decreased in the model group (P<0.01, P<0.05); and when compared with the model group, the mRNA expression of Rasd1, Greb1, Lgr6, Fras1 and Apod in embryonic tissues increased in the acupuncture group (P<0.05, P<0.01). These findings were consistent with the DEGs screening results. Gene ontology (GO) analysis revealed that the reversed genes of co-expression were primarily participated in the biological processes such as myoblast differentiation, endocardial cushion development, and cardiac morphogenesis. Kyoto encyclopedia of genes and genomes (KEGG) analysis indicated that the reversed genes of co-expression are enriched in the Wnt signaling pathway, and they were associated with various types of cardiac diseases. CONCLUSION The "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique may ameliorate the decline of ovarian response in mice with POR, promote normal follicle development and ovulation, thereby improve embryo quality and reduce the risk of developmental defects and deformity in the organs such as the embryonic heart. The underlying mechanism may be related to the regulation of the expression pattern of embryonic key genes through the Wnt signaling pathway.
Animals ; Female ; Mice ; Mice, Inbred C57BL ; Acupuncture Therapy ; Ovary/physiopathology* ; Humans ; Fertilization in Vitro ; Transcriptome ; Embryo Transfer ; Infertility, Female/physiopathology*

Nonobstructive azoospermia (NOA) is considered the most challenging clinical scenario for infertile men and current treatments leave many men unsuccessful at being able to achieve a pregnancy with their partner using their own sperm. Microdissection testicular sperm extraction (micro-TESE) is the choice for men with NOA desiring to father children with their own gametes. Micro-TESE results in the highest numbers of sperm cells retrieved for use with in vitro fertilization/intracytoplasmic sperm injection. With suboptimal micro-TESE success rates of sperm retrieval and then pregnancy and live birth using the retrieved sperm with in vitro fertilization/intracytoplasmic sperm injection, advances to improve outcomes are necessary. This article comprehensively reviews the technologies investigated to date to improve the outcomes for men undergoing micro-TESE.
Humans ; Male ; Sperm Retrieval ; Azoospermia/surgery* ; Pregnancy ; Female ; Sperm Injections, Intracytoplasmic ; Microdissection/methods* ; Fertilization in Vitro ; Testis/surgery*

Since the early days of assisted reproductive technology (ART), the importance of sperm processing, employed to separate the motile, morphologically normal sperm from the semen, has been shown to be beneficial. The aim of the semen processing technique has been to remove seminal plasma and facilitate capacitation. Additionally, the presence of leukocytes, bacteria, and dead spermatozoa has been shown to be detrimental as it may cause oxidative stress that has an adverse effect on oocyte fertilization and embryo development. Hence, removal of leukocytes, bacteria, and dead spermatozoa is an important step of sperm processing for assisted reproduction. Currently, several sperm processing techniques have been evolved and optimized in the field of assisted reproduction. The requirements for in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), and testicular sperm extraction (TESE) are different than those of intrauterine insemination (IUI). The yield of as many motile, morphologically normal sperm as possible is a prerequisite for the success of IVF insemination procedure. In ICSI, where injection of a single spermatozoon into the oocyte is performed by the embryologist, sperm selection techniques play a crucial role in the ICSI procedure. Finally, sperm retrieval in TESE samples with very low number of sperm may be challenging and requires extra care during sample processing. Additionally, sperm cryopreservation is necessary in TESE cases in order to avoid multiple biopsies.
Humans ; Sperm Injections, Intracytoplasmic/methods* ; Male ; Fertilization in Vitro/methods* ; Sperm Retrieval ; Andrology/methods* ; Cryopreservation ; Female ; Spermatozoa

The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoing in vitro fertilization (IVF) has been a debated hot topic among reproductive specialists. Each approach presents distinct advantages and disadvantages, with fresh sperm typically showing superior sperm quality, while frozen sperm offers logistical flexibility and a reliable backup for repeated cycles. This review summarizes the latest advancements in sperm retrieval and cryopreservation techniques, providing practitioners with a comprehensive analysis of each option's strengths and limitations. Comparative studies indicate that, although fresh sperm often has better quality metrics, cryopreservation methods such as vitrification have significantly improved postthaw outcomes, making frozen sperm a viable choice in assisted reproductive technologies (ART). The findings show comparable rates for fertilization, implantation, clinical pregnancy, and live birth between fresh and frozen microdissection testicular sperm extraction (micro-TESE) sperm in many cases, although patient-specific factors such as timing, cost-effectiveness, and procedural convenience should guide the final decision. Ultimately, the choice of using fresh or frozen sperm should align with the individual needs and conditions of patients. This tailored approach, supported by the latest advancements, can optimize ART outcomes and provide personalized reproductive care.
Humans ; Cryopreservation/methods* ; Male ; Sperm Retrieval ; Semen Preservation/methods* ; Azoospermia/therapy* ; Pregnancy ; Female ; Fertilization in Vitro ; Spermatozoa ; Microdissection ; Pregnancy Rate

OBJECTIVE: To study the durability of the anti-demineralization effects of fluoride varnish after being applied to dental root surfaces. METHODS: Coronal and radicular dentin samples were prepared from extracted human teeth. Duraphat^® (DP) was applied to the dentine surfaces to form a protective film. The film-dentin interfaces were observed by scanning electron microscopy (SEM) and the fluoride element was analyzed with energy dispersive spectrometer (EDS). Thus, the differences between applying DP on crowns and roots were compared. Radicular dentin samples were prepared and randomly divi-ded into four groups: (1) Blank: DP was not applied, and demineralized in acetic acid (pH 4.5) for 4 days; (2) Blank+aging: DP was not applied, the samples were put into deionized water for 14 days at room temperature, and then demineralized in acetic acid (pH 4.5) for 4 days; (3) DP: DP was applied and demineralized in acetic acid (pH 4.5) for 4 days; (4) DP+aging: DP was applied, the samples were put into deionized water for 14 days at room temperature, and then demineralized in acetic acid (pH 4.5) for 4 days. Finally, SEM observation and EDS analysis of fluoride content were performed on film-dentin interfaces to evaluate the degree of demineralization, the morphology of DP film, and the penetration of fluorine. RESULTS: The immediate penetration depth of fluoride element from DP was deeper in the coronal dentin than that in radicular dentin. The samples in the blank and blank+aging groups demine-ralized significantly after acid etching. The DP group did not undergo demineralization, and the fluorine element penetrated to (76.00±8.94) μm below the interfaces. The structure of the protective film in the DP+aging group was damaged, but the underneath dentin did not undergo demineralization. The fluorine element still remained at a depth of (5.00±3.53) μm below the interfaces. CONCLUSION DP has an anti-demineralization effect on the root surface, and this effect can still be exerted for a period of time after losing the structure of protective film. It has the ability to prevent root caries and a certain durability.
Humans ; Tooth Root/drug effects* ; Fluorides, Topical/pharmacology* ; Tooth Demineralization/prevention & control* ; Dentin/drug effects* ; Fluorides ; In Vitro Techniques ; Cariostatic Agents/pharmacology*

OBJECTIVE: To evaluate the filling quality of single cone obturation in root canal model with irregular structure (Hus&Kim Ⅴ, Yin Ⅱ-type isthmus) which established by 3D printing technology using slices and radiographic methods, in order to provide reference for clinical practice. METHODS: (1) Extracted fresh premolars with Hus&Kim Ⅴ and Yin-type Ⅱ isthmus were collected and scanned by cone-beam computed tomography (CBCT), then standard root canal models were designed and printed. Rhodamine B staining and bias fitting were used to verify the availability of the models. (2) 30 root canal models were randomly divided into 3 groups according to different filling methods (n=10). CONTROL GROUP: vertical compaction obturation; Experimental group 1: single cone obturation with 0.06-taper cone (30#); Experimental group 2: single cone obturation with 0.04-taper cone (35#), GuttaFlow 2 as canal sealers. Slices were taken at 2, 4, 6, and 8 mm from the root apex in the direction perpendicular to the long axis of the root and observed under a stereomicroscope to calculate the percentage of filling area (PAV), percentage of gutta-percha-filled area (PGFA), percentage of sealer filled area (PSFA). (3) On the basis of the above results, two groups (n=4) were selected to further analyze the filling quality by micro-computed tomography (Micro-CT), the filling volume of main root canal and the isthmus were obtained, and the percentage of filling volume (PFV) was calculated. Two-way ANOVA was used to evaluate the differences between the groups, and Tukey' s multiple comparison was used to compare the data between the groups and within the groups. RESULTS: (1) Rhodamine B staining solution could overflow the apical foramen, and the main root canal system and the isthmus area were stained, showed no remnants of support material. The 3D standard deviation of the printed model data was 0.03 mm, and the average fitting distance was 0.02 mm. (2) The PFA of the two experimental groups were both significantly lower than that of the control group (F=45.04, P < 0.01). There was no statistical difference of the PFA at apical 2 and 4 mm between the two experimental groups (P>0.01), but at the middle and coronal portions of the root canal (6, 8 mm), the PFA of the experimental group 1 was higher than that of the experimental group 2 (P < 0.01). PFA in the apical 2, 4 mm of the two experimental groups were both lower than that in the middle and coronal portions 6, 8 mm of the canal (P < 0.01). There was no difference in the PGFA and PSFA between the two experimental groups at the apical 2, 4 mm (F=2.383, P>0.01). (3) The results of Micro-CT showed that the PFV of the experimental group 1 was statistically different with the control group (F=47.33, P < 0.01). The PFV of the experimental group 1 was 54.33%±4.35% in the isthmus and 78.31%±4.21% in the main root canal, which were both lower than the PFV of the control group of 76.48%±4.89% (isthmus) and 86.90%±3.29% (main root canal, P < 0.01). The PFV of the main root canal in the experimental group 1 was higher than that in the isthmus (P < 0.01), while there was no difference between the isthmus and the main root canal in the control group (P>0.01). CONCLUSION In the irregular root canal structure with isthmus, using large-taper gutta-percha can improve the filling quality of the middle and upper part of the canal, but the percentage of filling volume in the isthmus is lower than that of the main canal, and more technical improvements are needed.
Humans ; Root Canal Obturation/methods* ; Cone-Beam Computed Tomography ; Root Canal Filling Materials ; Dental Pulp Cavity/diagnostic imaging* ; Printing, Three-Dimensional ; In Vitro Techniques ; Gutta-Percha ; Bicuspid

OBJECTIVES: Transcervical resection of adhesions (TCRA) under hysteroscopy is the mainstay treatment for intrauterine adhesions (IUA), but its effectiveness varies depending on the surgical approach. This study aims to investigate the impact of different surgical techniques on endometrial repair and pregnancy outcomes in patients with secondary infertility and moderate-to-severe IUA. METHODS: A retrospective analysis was conducted on 225 patients who underwent TCRA followed by in vitro fertilization and embryo transfer between January 2021 and December 2022. Patients were grouped based on the surgical method: A cold knife group (n=127) and an electrosurgical group (n=98). Adhesions were separated using either cold knife or electrosurgical instruments. Postoperative visualization of uterine angle and tubal ostia, endometrial restoration, vascular endothelial growth factor (VEGF) expression in adhesion tissues, and clinical pregnancy outcomes were compared. Univariate and multivariate Logistic regression analyses were performed to identify factors influencing pregnancy outcomes. A LightGBM model was constructed to predict pregnancy outcomes. RESULTS: Compared with the electrosurgical group, patients in the cold knife group had significantly greater postoperative endometrial thickness [(8.86±0.53) mm vs (8.10±0.87) mm, P<0.05], higher live birth rates (64.57% vs 30.61%, P<0.05), and lower VEGF expression (1.31±0.09 vs 1.53±0.16, P<0.05). Logistic regression analyses identified age, number of visible tubal ostia postoperatively, and surgical method as significant factors affecting pregnancy outcomes (P<0.05). The LightGBM model based on surgical method had an area under the curve (AUC) of 0.882 (0.838-0.926), with internal validation AUC of 0.817 (0.790-0.840). CONCLUSIONS Cold knife surgery promotes faster recovery of the endometrial microenvironment and earlier improvement of fertility in patients with secondary infertility and IUA Surgical method is a key factor influencing pregnancy outcomes, and the LightGBM model based on surgical approach shows good predictive performance for pregnancy outcomes in patients with moderate-to-severe IUA.
Humans ; Female ; Pregnancy ; Tissue Adhesions/surgery* ; Retrospective Studies ; Adult ; Pregnancy Outcome ; Uterine Diseases/surgery* ; Hysteroscopy/methods* ; Infertility, Female/etiology* ; Electrosurgery/methods* ; Fertilization in Vitro ; Endometrium/surgery* ; Embryo Transfer ; Vascular Endothelial Growth Factor A/metabolism*

OBJECTIVES: To explore the effects of exogenous trigger (hCG/GnRHa) versus endogenous LH surge in natural cycle IVF/ICSI (NC-IVF/ICSI) for patients with diminished ovarian reserve (DOR). METHODS: A retrospective analysis was conducted on 1,118 NC-IVF/ICSI cycles from two reproductive centers between 2013 and 2024. Propensity score matching (PSM) and multivariate logistic regression were used to adjust for confounding factors. The trigger-day hormone threshold was determined using receiver operating characteristic (ROC) curve analysis. Outcome measures included oocyte retrieval rate, 2PN fertilization rate, clinical available embryo rate, high-quality embryo rate, fresh cycle clinical pregnancy rate (CPR), and live birth rate (LBR). RESULTS: After adjusting for confounders via PSM and logistic regression, the exogenous trigger group demonstrated significantly better outcomes across all the evaluated parameters (oocyte retrieval rate, 2PN fertilization rate, transferable embryo rate, high-quality embryo rate, fresh cycle CPR, and LBR) than the endogenous LH surge group (P<0.05). Age-stratified analysis revealed that for the entire cohort, exogenous triggering significantly increased the number of transferable embryos and high-quality embryos (P<0.001). In the 35-39 years old subgroup, exogenous triggering showed significant advantages in oocyte yield, high-quality embryo rate, CPR, and LBR (P<0.05) and resulted in the most pronounced improvement in LBR (OR=6.25, 95% CI: 1.34-29.23). ROC analysis established a decision-day LH threshold of 19.055 mIU/mL (AUC=0.945, specificity=93.3%) for precise stratification of the clinical pathways. CONCLUSIONS For DOR patients undergoing NC-IVF/ICSI, exogenous triggering comprehensively improves the treatment outcomes, particularly providing significant live birth benefits for women aged 35-40 years. An individualized protocol incorporating the LH threshold (19.055 mIU/mL) effectively enhances embryonic developmental potential and live birth rates.
Humans ; Female ; Ovarian Reserve ; Pregnancy ; Propensity Score ; Retrospective Studies ; Fertilization in Vitro ; Sperm Injections, Intracytoplasmic ; Chorionic Gonadotropin ; Pregnancy Rate ; Logistic Models ; Ovulation Induction/methods* ; Gonadotropin-Releasing Hormone ; Adult ; Oocyte Retrieval

Some women may not be able to carry their own children even when capable of conceiving biological offspring. In-vitro fertilization and embryo-transfer (IVF-ET) through surrogacy can now make this possible for these women. Surrogacy however, is still considered unacceptable in the Philippines due to moral and legal issues. This article will explore the need and acceptability of surrogacy in this age of IVF-ET in a country where the prevailing social norms and religious values still disapprove of third-party assisted reproductive technology (ART). Medical indications that would benefit from gestational surrogacy were enumerated and briefly discussed. The differentiation between traditional and gestation surrogacy, as well as commercial and altruistic surrogacy were defined. IVF with gestational surrogacy is a feasible solution to a number of medical difficulties in the carrying of a gestation. Strictly regulating the practice and restricting its use only to cases with legitimate medical indications will prevent its misuse and exploitation. Moral issues, admittedly will still remain an issue particularly for commercial surrogacy. However, limiting these only to altruistic and gestational surrogacy in some cases may be an acceptable compromise.
Child ; Fertilization in Vitro ; Reproductive Techniques, Assisted

OBJECTIVE: To evaluate the effect of manual acupuncture on endometrial blood flow parameters by three-dimensional (3D) power Doppler ultrasound in women undergoing in vitro fertilization embryo transfer (IVF-ET). METHODS: Seventy patients undergoing IVF-ET were equally randomized into traditional or sham acupuncture treatment group for totally 4 days (from the day of oocyte aspiration to the day of embryo transfer) of treatment by random envelope method at the Reproductive Medicine Center and Outpatient Department of Integrated Traditional Chinese and Western Medicine, Tongji Hospital, Tongji Medicine College, Huazhong University of Science and Technology from January 2013 to December 2015. Patients in the traditional acupuncture group accepted traditional acupuncture methods with manual acupuncture, and Zhongji (CV3), Qihai (CV 6), Sanyinjiao (SP6), Taichong (LR 3), Tianshu (ST 25), Guilai (ST 29) and Zusanli (ST 36) were chosen. Patients at the sham acupuncture group accepted shallow acupuncture methods at 4 non-meridian points at each shoulder and upper arm. Outcome measures included endometrial ultrasonic indices such as vascularization index (VI), flow index (FI) and vascularization flow index (VFI), endometrial thickness and volume, subendometrial VI (sVI), subendometrial FI (sFI), subendometrial VFI (sVFI), implantation rate, clinical pregnancy rate, abortion rate, live birth rate and number of live births. RESULTS: Finally, 34 patients in the traditional acupuncture group and 35 in the sham acupuncture group completed this trial. VI, FI and VFI of the traditional acupuncture group were significantly higher than those in the sham acupuncture group (P<0.05). No significant differences were found in endometrial thickness, endometrial volume, sVI, sFI, sVFI, implantation rate, clinical pregnancy rate, abortion rate, live birth rate and number of live births (P>0.05). CONCLUSIONS Manual acupuncture performed after oocyte aspiration and before transplantation improved the endometrial blood flow parameters VI, RI and VFI in women who underwent IVF-ET, instead of sVI, sFI and sVFI. Therefore, acupuncture might be beneficial in women undergoing IVF-ET by increasing endometrial blood flow and endometrial receptivity. (Registration No. ChiCTR2100053354).
Pregnancy ; Humans ; Female ; Fertilization in Vitro/methods* ; Single-Blind Method ; Embryo Transfer ; Pregnancy Rate ; Acupuncture Therapy ; Endometrium/blood supply*