To investigate the therapeutic efficacy of artesunate (AS) on polycystic ovary syndrome (PCOS) in mice and explore the potential mechanism primarily.

Twenty-five female C57BL/6J mice were randomly divided into Control group, model group (PCOS group), low- and high-dose AS groups (AS15 and AS30 groups) and metformin group (Met group). In addition to the Control group, the mouse model of PCOS was established by subcutaneous injection of dehydroepiandrosterone (DHEA, 60 mg/kg) following by a high-fat diet for 21 d. After modeling, AS of 15 and 30 mg/kg was intraperitoneally injected into the mice of the AS15 and AS30 groups, respectively, and 200 mg/kg Met was given to those of the Met group by gavage, once per day, for 6 weeks. ELISA was used to detect serum testosterone (T), fasting insulin (FINS), luteinizing hormone (LH) and follicle-stimulating hormone (FSH), and the LH/FSH ratio was calculated. The levels of fasting blood glucose (FBG), triglyceride (TG) and total cholesterol (TC) were detected by automatic biochemical analyzer, and the homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. The estrous cycle was observed, and HE staining was performed for pathological changes in the ovary and uterus. Immunofluorescence assay was employed to measure the expression of p-eIF2α, ATF4 and CHOP in the ovarian tissue. After steroidogenic human granulosa-like tumor cell line KGN were exposed to 100 μmol/L DHEA to simulate the hyperandrogen environment of PCOS, and then treated with 5 and 10 μg/mL AS for 24 h, the protein levels of endoplasmic reticulum stress signaling pathway was detected by Western blotting.

Compared with the Control group, the PCOS mice had disturbed estrous cycle, polycystic changes in the ovaries, and significantly increased serum T level and LH/FSH ratio (P<0.05), and obviously elevated HOMA-IR, TC and TG levels in terms of metabolism (P<0.01). The expression levels of p-eIF2α, ATF4 and CHOP were notably up-regulated in the ovarian granulosa cells of PCOS mice and KGN cells after DHEA exposure (P<0.05). Additionally, AS treatment attenuated the pathological changes of ovary and uterine expression, decreased the serum T level and the LH/FSH ratio (P<0.05), and reduced HOMA-IR, TC and TG levels (P<0.05) when compared with the PCOS mice. Moreover, the expression levels of p-eIF2α, ATF4 and CHOP were significantly down-regulated after AS treatment in both ovarian granulosa cells of PCOS mice and KGN cells (P<0.05).

AS significantly improves glycolipid metabolic disorder and reproductive dysfunction in PCOS mice, which may be associated with its suppressing endoplasmic reticulum stress by inhibiting the PERK/eIF2α/ATF4/CHOP pathway.

Previous studies have suggested that vitamin D deficiency is common in patients with recurrent spontaneous abortion (RSA) and may be associated with immune-inflammatory imbalance and a prothrombotic state. In light of this, this study aims to investigate the relationship between serum 25-hydroxyvitamin D [25(OH)D] levels and inflammatory response as well as prothrombotic state in RSA patients.

A retrospective cohort study was conducted on 220 RSA patients who visited our reproductive medicine center between January 2022 and August 2023. Clinical data were analyzed, and serum levels of 25(OH)D, inflammatory markers (IL-6, TNF-α, IL-10, hsCRP), prothrombotic state indicators [total homocysteine (tHCY), D-dimer], glucolipid metabolism parameters, thyroid function, immune cell subsets and ovarian reserve function were assessed. According to 25(OH)D levels, these RSA patients were stratified into a severe deficiency group (<10 ng/mL, n=14), a deficiency group (10 to <20 ng/mL, n=140), an insufficiency group (20 to <30 ng/mL, n=53), and a sufficiency group (≥30 ng/mL, n=13). The abovementioned indicators were compared among the groups. Mann-Whitney U test was used for comparisons between 2 groups. For comparisons among more than 2 groups, Kruskal-Wallis H test was performed followed by Dunn’s post hoc test. Spearman and partial correlation analyses were applied to evaluate associations between 25(OH)D and related indicators.

Vitamin D deficiency or insufficiency was observed in 94.09% of the cohort. Compared with the patients with 25(OH)D ≥20 ng/mL, those with levels <20 ng/mL had significantly higher tHCY levels [8.35 (7.23 to 9.60) μmol/L vs 7.60 (6.35 to 8.95) μmol/L, P=0.007]. No statistical differences were observed in the intergroup comparisons in glucolipid metabolism, thyroid function, inflammatory markers, immune cell subsets or ovarian reserve function. Spearman correlation analysis showed that serum 25(OH)D levels were negatively correlated with tHCY (r=-0.222, P<0.01), and this association remained significant after adjusting for age and body mass index (r=-0.188, P=0.019). In the patients with 25(OH)D <30 ng/mL, a weak negative correlation was observed between 25(OH)D and IL-6 (r=-0.159, P<0.05), but this difference was not statistically significant after adjustment.

Vitamin D deficiency is quite common in RSA patients. Lower 25(OH)D levels are associated with an altered prothrombotic profile, particularly elevated tHCY, whereas their association with inflammatory markers appears limited. These findings suggest that vitamin D status may be involved in the regulation of thrombotic risk associated with RSA.

To investigate the effect of insulin resistance (IR) in primary infertile couples on pregnancy outcomes in fresh cycle embryo transfer.

A total of 101 primary infertile couples who underwent first IVF-ET/ICSI assisted conception with fresh cycle embryo transfer in our reproductive medical center from October 2022 to October 2023 were retrospectively included. According to whether they had combined IR unilaterally, these couples were divided into female IR group (n=49) and female non-IR group (n=52), and male IR group (n=49) and male non-IR group (n=52). The basic data, ovulation promotion, embryo transfer and pregnancy outcome were compared between the female IR group and non-IR group, and between the male IR group and non-IR group. Logistic regression analysis was applied to determine the effect of IR on pregnancy outcome and laboratory indicators of ovulation promotion.

①The IR female group had significantly higher body weight and BMI, and lower antral follicle count (AFC) when compared with the non-IR female group (P<0.05). ②Female fasting blood glucose was negatively correlated with the number of acquired eggs, number of metaphase Ⅱ (MII) oocytes, number of transferable embryos, and number of high-quality embryos (P<0.05); and both female fasting insulin level and female HOMA-IR were positively correlated with the days and total amount of gonadotropin (GN) (P<0.05). ③Embryo implantation and clinical pregnancy rates were significantly lower in the female IR group than the female non-IR group (P<0.05). ④The male IR group had significantly higher BMI than the male non-IR group (P<0.05). ⑤Embryo implantation rate and clinical pregnancy rate were in declining trend in all following 3 groups of couples (both IR groups, single IR group, and neither IR group). ⑥Female fasting blood glucose level showed a negative effect on number of acquired eggs, number of MII oocytes, number of transferable embryos, and number of high-quality embryos.

IR affects embryo quality in IVF/ICSI cycles in women with primary infertility; IR in couples with primary infertility negatively affects embryo implantation rates and clinical pregnancy rates in IVF/ICSI cycles.