ObjectiveTo investigate the protective effects and mechanisms of Bushen Zhuyun prescription （BSZY） on abortion rats with kidney deficiency-corpus luteum inhibition syndrome. MethodsAn abortion rat model with kidney deficiency-corpus luteum inhibition syndrome was constructed. Pregnant mice aged 8-10 weeks were randomly divided into a control group （Control）， a model group （Model）， low-dose BSZY （BSZY-L）， medium-dose BSZY （BSZY-M）， and high-dose BSZY （BSZY-H） groups （2.57， 5.14， 10.28 g·kg^-¹）， and a Zishen Yutai Pill （ZSYT） group （1.575 g·kg^-¹）. Hematoxylin-eosin （HE） staining was used to evaluate histopathological changes in ovarian and decidual tissue of rats in each group. Enzyme-linked immunosorbent assay （ELISA） was employed to measure levels of estrogen （E₂）， progesterone （P）， luteinizing hormone （LH）， prolactin （PRL）， and follicle-stimulating hormone （FSH） in serum. The candidate targets of BSZY were obtained from the Traditional Chinese Medicine System Pharmacology Platform （TCMSP） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP） v2.0 databases， while disease targets for recurrent spontaneous abortion （RSA） were retrieved from GeneCards， DrugBank， Online Mendelian Inheritance in Man （OMIM）， and Therapeutic Target Database （TTD）. The intersection targets were identified by the Venny 2.1.0 platform. Pathway enrichment analysis was conducted based on the Metascape database to predict the potential mechanisms of BSZY. Additionally. Western blot was used to verify the effects of BSZY on the expression of estrogen receptor （ERα）， phosphatidylinositol 3-kinase （PI3K）， and protein kinase B （Akt） and explore its protective mechanism on RSA rats. ResultsCompared with the control group， the model group exhibited significantly decreased uterine， ovarian， and embryonic wet weights （P<0.05， P<0.01）， with an abortion rate of 57.18%. The ovarian tissue showed varying degrees of reduction in primordial follicles， primary follicles， mature follicles， and corpora lutea， along with a large number of atretic follicles. The endometrium was thinner， and decidual tissue exhibited cellular edema and disorganized arrangement. In contrast， compared with the model group， the BSZY groups at all doses and the ZSYT group demonstrated increased uterine， ovarian， and embryonic wet weights， along with a reduced abortion rate. The number of primordial follicles， primary follicles， mature follicles， and corpora lutea increased， while atretic follicles decreased. The endometrium thickened， and decidual tissue displayed normal cellular structure with tight arrangement. Additionally， the model group showed significantly decreased levels of E₂， P， PRL， and FSH in serum （P<0.05， P<0.01）， along with a decreasing trend in LH level. In contrast， the BSZY groups at all doses exhibited significantly elevated levels of E₂， P， LH， PRL， and FSH in serum （P<0.05， P<0.01）. Network pharmacology predictions suggested that BSZY may exert protective effects against abortion in rats by activating the ERα/PI3K/Akt signaling pathway. Western blot results confirmed that BSZY significantly upregulated the expression of ERα， PI3K， and p-Akt proteins （P<0.05， P<0.01）. ConclusionBSZY has a protective effect on the abortion rats with kidney deficiency-corpus luteum inhibition syndrome， possibly by activating the ERα/PI3K/Akt signaling pathway to reduce ovarian apoptosis and regulate endocrine function， thereby lowering the abortion rate.

“Deficiency cause， cold accumulation， and qi stagnation” originates from Essentials from the Golden Cabinet （《金匮要略》）， which is a guiding principle for the pathogenesis of women's diseases， pioneering the differentiation and treatment of women's diseases based on patterns， and having a profound influence on future generations. Following the classical principles and simplifying the complexities， this paper explored the pathogenesis and mechanism of polycystic ovary syndrome （PCOS） from the perspective of “deficiency cause， cold accumulation， and qi stagnation”， and believed that depletion of essence and blood， long-term accumulation of internal cold， and qi constraint and blood stasis are the causes of PCOS， with depletion of essence and blood， and lack of nourishment of zang-fu （脏腑） organs as the root， and cold pathogen invasion， qi constraint and blood stasis as the branch. The main treatment principle is “treating deficiency with supplementation”， and dispelling pathogen while reinforcing healthy qi， along with “treatment of cold by warming” and “treatment of stagnation by dispersing”. This is of great significance for the treatment of polycystic ovary syndrome. Clinically， these methods can be used flexibly to guide treatment and formula selection for PCOS， with the goal of harmonizing qi and blood and regulating menstruation.

Objective To summarize and evaluate the characteristics of current recurrent spontaneous abortion(RSA)animal models at home and abroad,and to provide reference and guidance for the standardized preparation of RSA models.Methods"Recurrent spontaneous abortion"and"animal model"were used as co-keywords in CNKI,Wanfang,VIP,PubMed and Web of Science databases to search the RSA animal experimental literature,covering the period up to January 20,2024,and a total of 1 411 articles were collected.The analysis focused on construction methods and essential elements of RSA animal models,the modeling process and result evaluation,as well as the application of these models in pharmacological and pharmacodynamic research.An Excel table was established for systematic analysis and discussion.Results A total of 138 experimental studies were obtained after screening.In constructing RSA animal models,immunological models were the most widely used in Western medicine(96.92％),with the Clark model being the main one(92.31％).In traditional Chinese medicine(TCM)models,70.00％were kidney deficiency-luteal inhibition-syndrome combination models,20.00％were kidney deficiency and blood stasis models,and 10.00％were deficiency-heat syndrome models.Most animals were selected at 6-8 weeks(33.86％)and 8 weeks(32.28％)of age.The majority of animals were paired for mating at 18:00 on the day of cage pairing.In 81.03％of literatures,vaginal plugs were checked once the following morning,with 8:00 being the most common time(17.02％).The most commonly used drug administration cycle was 14 days of continuous gavage after pregnancy.Among the tested drugs,Western drugs were mainly protein-based(29.17％),while TCM drugs were mainly TCM decoction(81.11％).The most frequently used methods for detecting indicators included visual observation of embryos(22.54％),western blot(15.96％),PCR(13.58％),ELISA(12.91％),HE staining(10.80％)and immunohistochemistry(9.39％).Conclusion The etiology of RSA is complex,and corresponding animal models should be established based on different etiologies.Clark model is commonly used in the construction of Western medicine model,while the kidney deficiency-luteal inhibition-syndrome combination model is predominant in TCM.RSA animal model is widely used in related research,but systematic evaluation needs to be strengthened.

Vaccination is the most effective measure for reducing and preventing influenza and related complications. In this study, we analyzed the mutation trend and the antigen dominant site changes of the amino acid sequence of hemagglutinin subunit 1 (HA1) of human influenza A virus (IAV) in the northern hemisphere from 2012 to 2022. According to the HA1 sequences of A/Darwin/6/2021 (H3N2) and A/Wisconsin/588/2019 (H1N1) recommended by the World Health Organization in the 2022 influenza season in northern hemisphere, we employed the mosaic algorithm to design three Mosaic-HA1 antigens through stepwise substitution. Mosaic-HA1 was expressed and purified in 293F cells and then mixed with the alum adjuvant at a volume ratio of 1:1. The mixture was used to immunize BALB/c mice, and the immunogenicity was evaluated. Enzyme-linked immunosorbent assay showed that Mosaic-HA1 induced the production of IgG targeting two types of HA1, the specific IgG titers for binding to H3 protein and H1 protein reached 10⁵ and 10³ respectively. The challenge test showed that Mosaic-HA1 protected mice from H3N2 or H1N1. This study designs the vaccines by recombination of major antigenic sites in different subtypes of IAV, giving new insights into the development of multivalent subunit vaccines against influenza.
Animals ; Influenza A Virus, H1N1 Subtype/genetics* ; Influenza A Virus, H3N2 Subtype/genetics* ; Mice, Inbred BALB C ; Mice ; Influenza Vaccines/genetics* ; Hemagglutinin Glycoproteins, Influenza Virus/genetics* ; Humans ; Antibodies, Viral/blood* ; Antigens, Viral/genetics* ; Immunoglobulin G/immunology* ; Female ; Orthomyxoviridae Infections/prevention & control* ; HEK293 Cells

Objective To construct a mutant strain of Nocardia farcinica ( N. farcinica ) IFM10152 with mammalian cell entry 4A gene (mce4A) deletion and to analyze the function of that gene dur-ing infection. -ethods The mutant strain of N. farcinica was constructed through in-frame deletion without antibiotic labeling and verified by PCR and sequencing analysis. To analyze the function of mce4A gene in the interaction between N. farcinica and host cells, in vitro growth experiment, macrophage killing experi-ment using THP-1 ( a human leukemia mononuclear cell line) as the model and adhesion and invasion exper-iments using HeLa cells ( cervical cancer epithelial cells) were carried out. Results The mutant strain with mce4A gene deletion was successfully constructed and named △mce4A. No significant difference in growth rate was observed between the mutant and the wild-type strains. After knocking out the mce4A gene, the ability of N. farcinica to resist macrophage killing was obviously weakened as well as its ability to adhere and invade. Conclusions The mutant strain of N. farcinica with mce4A gene deletion was successfully construc-ted. The mce4A gene might play an important role in the adhesion and invasion of N. farcinica to host cells and its survival in macrophages.

Objective To investigate the effects of high glucose and lysophosphatidylcholine (LPC) on the immune function of in vitro cultured macrophages during Nocardia farcinica infection. Meth-ods RAW264.7 macrophages were cultured in vitro under different conditions as follows: routine culture (control group),50 mmol/L glucose (high glucose group),10 mg/L LPC(LPC groupⅠ),25 mg/L LPC (LPC groupⅡ) and 50 mmol/L glucose+25 mg/L LPC(high glucose and LPC group). The activity of mac-rophages in each group was tested after 6,12,24 and 36 h of culture. After 24 h of culture, macrophages were collected from every group and co-cultured with Nocardia farcinica. Dynamic phagocytosis rates were detected at 1,2,3,4,5 and 6 h after co-culture. Toxic effects of Nocardia farcinica on macrophages and concentrations of IL-10 and TNF-α were measured at 1,3 and 6 h after co-culture. Results Macrophages in all four experimental groups showed decreased activity as compared with those in the control group (P<0.01). Phagocytosis of Nocardia farcinica by macrophages was also reduced by high glucose and LPC. Phagocytosis rates of high glucose group and LPC groupⅡ at 1 and 2 h,LPC groupⅠat 1,2 and 3 h,and high glucose and LPC group at 1,2,3 and 4 h after co-culture were significantly lower than that of the con-trol group (P<0.05 or P<0.01). Compared with the control group, significantly reduced toxic effects on macrophages caused by Nocardia farcinica was observed in the experimental groups (P<0.05 or P<0.01). Compared with the control group,LPC groupsⅠand Ⅱ and high glucose and LPC group had decreased se-cretion of IL-10 at 3 h,and high glucose group and LPC groupⅠhad decreased secretion of TNF-α at 1 h(P<0.05). Conclusion Culture macrophages under the conditions of high glucose and LPC would reduce their activity and impair their ability to phagocytose Nocardia farcinica. Moreover, high glucose and LPC might have impacts on the toxic effects of Nocardia farcinica on macrophages and the secretion of IL-10 and TNF-α.