[Effect of Yunkang Oral Liquid on preventing LPS-induced abortion and regulating immune tolerance in mice].

To investigate the effect of Yunkang Oral Liquid on preventing lipopolysaccharide(LPS)-induced abortion and regulating immune tolerance in mice,sixty normal ICR mice were randomly divided into normal group,model group,Yunkang Oral Liquid high,middle and low dose groups and progesterone group.Abortion model was established by tail vain injection of LPS(0.1µg/mouse)on the 7th day of pregnancy.Since the first day of pregnancy,the same volume of distilled water,Yunkang Oral Liquid at the dose of 36,18 and 9 m L·kg~(-1)·d~(-1),and progesterone at the dose of 0.038 g·kg~(-1)·d~(-1)were given in corresponding groups.The mice were sacrificed at the 9th day of pregnancy,and the embryo loss of each group was calculated.The levels of Th1 type cytokines(TNF-α,IFN-γ)and Th2 type cytokines(IL-4,IL-10)in uterus homogenate were detected by ELISA.HE staining was performed to examine the histopathological changes in the decidua.The expression levels of CD14,F4/80 in macrophages of uterus were detected by immunohistochemistry.Western blot was used to investigate the protein expression of TLR4,MyD88 and NF-κB in uterine decidua.In our study,all Yunkang Oral Liquid groups could significantly reduce the embryo absorption rate of mice,while high dose group can significantly increase the levels of IL-10 and IL-4;both medium and high dose groups can significantly decrease TNF-α,and IFN-γlevelsin the uterus of model mice,reduce the protein expression of NF-κB,MyD88 and TLR4 in uterine decidua tissue.Various treatment groups could reduce the counts of F4/80,CD14 macrophages and decrease expression area in uterine tissue.Our results showed that Yunkang Oral Liquid could prevent LPS-induced abortion,and the mechanism may be associated with inhibiting the TLR4/MyD88/NF-κB signaling pathway and regulating the balance of Th1/Th2 immune factors,which could improve the endometrial receptivity of mice,and promote the development of decidua and implantation of embryo.

[Antihypertensive effect and mechanism of Dendrobium officinale flos on high-blood pressure rats induced by high glucose and high fat compound alcohol].

This study aimed to investigate the antihypertensive effect and possible mechanism of Dendrobium officinale flos on hypertensive rats induced by high glucose and high fat compound alcohol. The hypertensive models were successfully made by high-glucose and high-fat diet, with gradient drinking for 4 weeks, and then divided into model control group, valsartan (5.7 mg·kg⁻¹) positive control group and D. officinale flos groups (3，1 g·kg⁻¹). After 6 weeks of treatment, the blood pressure of rats was measured regularly. After the last administration, endothelin-1 (ET-1), thromboxane B2 (TXB2), prostacyclin (PGI2) and nitric oxide (NO) were tested. Endothelial nitric oxide synthase (eNOS) expression and lesion status in thoracic aorta were detected. The vascular endothelium dependent dilation of the thoracic aorta was detected by the isolated vascular loop tension test. The results showed that D. officinale flos could significantly reduce systolic blood pressure and mean arterial pressure in hypertensive rats, inhibit the thickening of thoracic aorta and the loss of endothelial cells, reduce plasma content of ET-1 and TXB2, and increase the content of PGI2 and NO. After long-term administration, vascular endothelium dependent dilation of the thoracic aorta was significantly increased, and could be blocked by the eNOS inhibitor (L-NAME) and increase the expression of eNOS. Therefore, D. officinale flos has an obvious antihypertensive effect on high glucose and high fat compound alcohol-induced hypertensive rats. Its mechanism may be correlated with the improvement of vascular diastolic function by protecting vascular endothelial cells, and finally resist hypertension.

[Effect and mechanism of Yunkang oral liquid in regulating endocrine system and VEGF signaling pathway and reducing abortion rate in recurrent abortion mice].

This experiment focuses on the effect of Yunkang oral liquid on abortion rate, endocrine system and VEGF signal pathway in Clark classical recurrent abortion model mice. RSA mice were randomly divded into model group, low, middle and high-dose groups and progesterone group. The normal pregnancy mice were included into normal group. Since the first day of pregnancy, the normal group and the RSA model group were given the same dose of distilled water, while low, middle and high-dose groups were given Yunkang oral liquid at the dose of 9, 18, 36 mL·kg¹·d⁻¹; progesterone group were given progesterone by 0.039 g·kg¹·d⁻¹. The mice were put to deathat the 15th day of pregnancy, and the embryo loss rate of each group was observed. Serum estradiol (E2), progesterone (P), prolactin (PRL), luteinizing hormone (LH), follicle stimulating hormone (FSH) level were tested; the protein expressions of estrogen receptor(ER), progesterone receptor (PR), prolactin receptor (PRLR) in decidua and RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in deciduas were studied. The results showed that middle, high dose Yunkang and progesterone could significantly decrease the embryo loss rate of RSA mice. The levels of FSH, LH, PRL, P and E2 in serum in Yunkang and progesterone groups were increased, and the serum levels of FSH, LH, and E2 in Yunkang group were higher than those in progesterone group. Western blot analysis showed that Yunkang oral liquid and progesterone can significantly increase the expressions of PRLR, PR in the uterine decidua of RSA mice, and the expression of ER in Yunkang group was higher than that in progesterone group. Western blot and PCR showed that the Yunkang oral liquid and progesterone can significantly increase RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in the uterine decidua of RSA mice. The results showed that Yunkang oral liquid can effectively reduce the embryo loss rate of RSA model mice, increase the levels of FSH, LH, PRL, P and E2 in serum, promote the expressions of PRLR, PR, ER protein in decidua and the RAS, MAPK, VEGF, VEGFR-2 gene and protein expressions in the decidua, improve the vascular remodeling of fetal interface, the endometrial receptivty, the development of decidua and the blastocyst implantation.

[Efficacy of compound Dendrobium on PI3K/AKT/eNOS signaling pathway in hypertensive rats induced by "dietary disorders"].

To observe the efficacy of compound Dendrobium on Sprague Dawley rats (SD) hypertension model induced by "dietary disorders" and its relevant mechanism, totally 50 SD rats were fed with high-sugar, high-fat diet and alcohol for four weeks. According to the blood pressure after modeling, the rats were divided into model group, valsartan group (8 mg·kg⁻¹), low, medium and high-dose Dendrobium candidum compound groups (1.65, 3.30, 5.00 g·kg⁻¹), with 10 rats in each group, and the other 10 SD rats were also taken as the normal group. After four weeks of treatment, blood pressure was measured. Orbital blood was collected for the determination of serum cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL), calculation of atherosclerosis index (AI). Nitric acid reductase method was used to detect serum nitric oxide (NO); the levels of serum endothelin-1 (ET-1) and intercellular adhesion molecule-1 (ICAM-1) were measured by ELISA. The rats were put to death after the last administration, and the protein expressions of PI3K/AKT/eNOS in thoracic aorta of rats in each group were detected by Western blot. The aorta was separated and stained with hematoxylin-eosin (HE) to observe the changes in the endothelium and blood vessels in the thoracic aorta. Masson staining was used to observe the formation of aortic collagen. The expressions of nitric oxide synthase (eNOS) and ICAM-1 in aortic endothelial cells were observed by immunohistochemistry. In contrast, the results show D. candidum compound can significantly reduce the blood pressure in hypertensive rats, increase HDL-c, and reduce AI, while increasing serum NO content, decreasing ET-1 and ICAM-1 levels and promoting PI3K/AKT/NOS protein expressions. The lesion degree of the D. candidum compound group was reduced, and the collagen deposition was significantly reduced. Meanwhile, D. candidum compound can significantly increase the expression of eNOS, and reduce the formation of ICAM-1.Therefore, D. candidum compound has an obvious antihypertensive effect on hypertensive rats, which may be related to the increase in PI3K/AKT/eNOS signaling pathways and NO generation, the inhibition of the secretion of ICAM-1 and ET-1, the protection of the vascular endothelium and the improvement of aortic disease.

Discovery of orally bioavailable phenyltetrazolium derivatives for the acute treatment and the secondary prevention of ischemic stroke.

The potential for secondary stroke prevention, which can significantly reduce the risk of recurrent strokes by almost 90%, underscores its critical importance. N-butylphthalide (NBP) has emerged as a promising treatment for acute cerebral ischemia, yet its efficacy for secondary stroke prevention is hindered by inadequate pharmacokinetic properties. This study, driven by a comprehensive structural analysis, the iterative process of structure optimization culminated in the identification of compound B4, which demonstrated exceptional neuroprotective efficacy and remarkable oral exposure and oral bioavailability. Notably, in an in vivo transient middle cerebral artery occlusion (tMCAO) model, B4 substantially attenuated infarct volumes, surpassing the effectiveness of NBP. While oral treatment with B4 exhibited stronger prevention potency than NBP in photothrombotic (PT) model. In summary, compound B4, with its impressive oral bioavailability and potent neuroprotective effects, offers promise for both acute ischemic stroke treatment and secondary stroke prevention.

Rational Identification of Novel Antibody-Drug Conjugate with High Bystander Killing Effect against Heterogeneous Tumors.

Bystander-killing payloads can significantly overcome the tumor heterogeneity issue and enhance the clinical potential of antibody-drug conjugates (ADC), but the rational design and identification of effective bystander warheads constrain the broader implementation of this strategy. Here, graph attention networks (GAT) are constructed for a rational bystander killing scoring model and ADC construction workflow for the first time. To generate efficient bystander-killing payloads, this model is utilized for score-directed exatecan derivatives design. Among them, Ed9, the most potent payload with satisfactory permeability and bioactivity, is further used to construct ADC. Through linker optimization and conjugation, novel ADCs are constructed that perform excellent anti-tumor efficacy and bystander-killing effect in vivo and in vitro. The optimal conjugate T-VEd9 exhibited therapeutic efficacy superior to DS-8201 against heterogeneous tumors. These results demonstrate that the effective scoring approach can pave the way for the discovery of novel ADC with promising bystander payloads to combat tumor heterogeneity.

Injectable and Sprayable Fluorescent Nanoprobe for Rapid Real-Time Detection of Human Colorectal Tumors.

The development of minimally invasive surgery has greatly advanced precision tumor surgery, but sometimes suffers from restricted visualization of the surgical field, especially during the removal of abdominal tumors. A 3-D inspection of tumors could be achieved by intravenously injecting tumor-selective fluorescent probes, whereas most of which are unable to instantly distinguish tumors via in situ spraying, which is urgently needed in the process of surgery in a convenient manner. In this study, we have designed an injectable and sprayable fluorescent nanoprobe, termed Poly-g-BAT, to realize rapid tumor imaging in freshly dissected human colorectal tumors and animal models. Mechanistically, the incorporation of γ-glutamyl group facilitates the rapid internalization of Poly-g-BAT, and these internalized nanoprobes can be subsequently activated by intracellular NAD(P)H: quinone oxidoreductase-1 to release near-infrared fluorophores. As a result, Poly-g-BAT can achieve a superior tumor-to-normal ratio (TNR) up to 12.3 and enable a fast visualization (3 min after in situ spraying) of tumor boundaries in the xenograft tumor models, Apcmin/+ mice models and fresh human tumor tissues. In addition, Poly-g-BAT is capable of identifying minimal premalignant lesions via intravenous injection. This article is protected by copyright. All rights reserved.

Design, synthesis and biological evaluation of sulfonamides inhibitors of XPO1 displaying activity against multiple myeloma cells.

Multiple myeloma (MM) is a highly malignant hematologic cancer that occurs when an atypical plasma cell develops in the bone marrow and reproduces quickly. Despite varies of new drugs have been developed or under clinic trial, MM is still essentially incurable, while XPO1 inhibition has emerged as a promising therapeutic strategy in the treatment of MM. Using the second-generation XPO1 inhibitor KPT-8602 as the lead compound, structure-based optimization provided D4 with high anti-proliferation efficacy (IC50 = 24 nM in MM.1S). In addition, the treatment with D4 significantly induced MM.1S cell cycle arrested and cell apoptosis, which was confirmed as on-target effect by immunofluorescence microscopy and competitive binding assay. Moreover, D4 displayed good metabolic stability over rat plasma and liver microsomes, as well as good pharmacokinetic profile on SD rat model with high drug exposure and decent bioavailability by oral gavage. All these good properties of D4 pave the way for further drug development and clinical application.

Discovery of seneciobipyrrolidine derivatives for the amelioration of glucose homeostasis disorders through 4E-BP1/Akt/AMPK signaling activation.

Modulating the glucose transport in skeletal muscle is a promising strategy for ameliorating glucose homeostasis disorders. However, the complicated mechanisms of glucose transport make it difficult to find compounds therapeutically relevant molecular mechanisms of action, while phenotypic screening is thought to be an alternative approach to mimic the cell state of interest. Here, we report (±)-seneciobipyrrolidine (1a) is first found to enhance glucose uptake in L6 myotubes through phenotype-based screening. Further SAR investigation led to the identfication of compound A27 (EC50 = 2.7 µM). Proteomiic analysis discloses the unique function mechanism of A27 through upregulating the level of the eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), subsequently enhancing the Akt and AMPK phosphorylation, thereby promoting the glucose uptake. Chronic oral administration of A27 significantly lowers blood glucose and improves glucose tolerance in db/db mice. This work is new research on seneciobipyrrolidine derivatives, providing a promising avenue for ameliorating glucose homeostasis.