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.

To screen the targets related to the metabolic enzymes involved in the cholesterol synthesis pathway that inhibits the polarization of macrophages towards M1 phenotype, and verify the intervention effects and underlying mechanisms in colorectal cancer cells.

Mouse colorectal cancer MC38 cells were divided into control group (si-NC) and experimental groups (the expression of enzymes in cholesterol synthesis pathway was interfered with siRNA for corresponding targets). RT-qPCR was used to detect the mRNA levels of corresponding targets in MC38 cells after transfection. After peritoneal macrophages were extracted from male C57BL/6 mice (6 weeks old, weighing 13~18 g), the macrophages were then treated with the conditioned media of MC38 cells transfected with different siRNAs for 48 h. RT-qPCR was employed to detect the mRNA levels of IL-1β, IL-6 and TNF-α in the macrophages so as to evaluate the effect of the culture media on the M1 polarization. MC38 cells were divided into control groups (OE-NC and sh-NC), farnesyl-diphosphate farnesyltransferase 1 (FDFT1) overexpression group (OE-FDFT1) and FDFT1 knockdown group (sh-FDFT1). RT-qPCR was applied to detect the mRNA expression of FDFT1, and Western blotting was conducted to measure the protein level of FDFT1. C57BL/6 mice were subjected randomly to construct a subcutaneous tumor-bearing model and a model of intraperitoneal metastatic tumor (n=5) respectively. The growth of tumor mass was then measured. Flow cytometry was used to observe the proliferation and apoptosis of MC38 cells, and Transwell assay to detect migration ability of MC38 cells. Five C57BL/6 macrophage-depleted mice (established with injection of clodronate liposome suspension through tail vein) received intraperitoneal implantation to construct a metastasis model, and then the obtained tumor masses were then weighted.

Compared with MC38 cells after si-NC transfection, the mRNA levels of corresponding targets in MC38 cells in the experimental groups were significantly reduced (P<0.05). Significant increases were found in the mRNA levels of IL-1β, IL-6 and TNF-α of the macrophages with FDFT1 interference than the control cells (P<0.05). There were no statistical differences in the proliferation, apoptosis and migration of MC38 cells in the control group (OE-NC and sh-NC) and the cells of the FDFT1 overexpression group and FDFT1 knockdown group (P>0.05). In both the subcutaneous tumor-bearing model and the model of intraperitoneal metastatic tumor, the mass weight was significantly heavier in the OE-FDFT1 group than the OE-NC group (P<0.01), and was notably smaller in the sh-FDFT1 group than the sh-NC group (P<0.01). For the macrophage-depleted mouse tumor model, no remarkable change was observed in the tumor weight between the OE-FDFT1 group and the OE-NC group as well as the sh-FDFT1 group and the sh-NC group.

FDFT1, the metabolic enzyme in the cholesterol synthesis pathway of colorectal cancer tumor cells, is a potential target for tumor immunotherapy targeting macrophages, which promotes tumor progression by regulating macrophages.

To explore the role and mechanism of warm malate ringer’s solution (MR) in resuscitation of the lethal triad caused by severe trauma.

A rat model of severe trauma was established in SPF-grade SD rats (half male and half female, weighing 200~220 g) using combined multiple injuries and hemorrhagic shock, and the rats were randomly divided into 8 groups (n=8): Sham group, only arterial and venous catheterization; Trauma (Tra) groups with different time points (10, 30, 60, 90, 120, 180 min) and a Trauma group that were observed without any treatment for 180 min after model establishment. The changes of activated clotting time (ACT), reaction time (R), maximum amplitude (MA), and rate of blood clot formation (Angle) at different time points were detected by using thromboelastography, and tail bleeding, core body temperature and arterial blood gas parameters, were also observed and detected. The plasma von Willebrand Factor (vWF) level, mitochondrial respiratory control ratio in pulmonary venous endothelium, and expression levels of vascular endothelial cadherin (VE-Cadherin), peroxisome proliferator activating receptor gamma coactivator 1α (PGC1α), dynamin-related protein 1 (Drp1), p-Drp1, and mitofusin 2 (Mfn2) were detected to evaluate the vascular endothelial injury and mitochondrial dysfunction. Another group of SD rats were randomly divided into severe trauma group (no treatment for 180 min after injury), and MR solution at room temperature and at 37 ℃ groups. MR solution at room temperature or at 37 ℃ was given to the rats using a medical blood transfusion apparatus at 60 min post-trauma. Above indicators were observed and detected to investigate the resuscitation effect of the MR solution.

Compared with the Sham group, the severely traumatic rats at 180 min after injury had significantly prolonged ACT and R values (P<0.05), shortened MA and decreased Angle values (P<0.05), extended tail bleeding time (P<0.05), lower partial pressure of carbon dioxide (PCO₂) and HCO{}_{\mathrm{3}}^{-} and base excess (BE) levels (P<0.05), and continuously increasing K⁺ (P<0.05) and decreasing Na⁺ (P<0.05) and Ca²⁺ levels (P<0.05). Additionally, plasma vWF level (P<0.05) and protein levels of VE-cadherin, PGC1α and Mfn2 in pulmonary vein endothelium were significantly reduced (P<0.05), the expression of p-Drp1 was enhanced and the mitochondrial respiration control rate was declined in the rats at 180 min after injury (P<0.05). MR solution resuscitation shortened tail bleeding time (P<0.05), increased core body temperature (P<0.05), elevated plasma vWF level (P<0.05), increased protein levels of VE-cadherin, PGC1α and Mfn2 (P<0.05), and decreased that of p-Drp1 protein expression (P<0.05) when compared with the rats at 180 min after severe traumatic injury. The above effects were more significant in the rats infused with the solution at 37 ℃ than those at room temperature.

Warm MR solution significantly improves the lethal triad in rats after severe trauma, which may be associated with its improving mitochondrial function and attenuating vascular endothelial damage.

‍To investigate the effect of dexmedetomidine （Dex） on bone loss in tail-suspended rats and primarily explore its regulatory mechanism on bone metabolism.

‍A total of 30 male rats were randomly divided into a control group，a model group，and a Dex group，with 10 animals in each group. Rat model of osteoporosis was established by hind limb suspension for 4 weeks. Dex at a dose of 10 μg/kg was given intraperitoneally，once every other day from the day of tail suspension. And equal amount of normal saline was given to the control and model group. Bone histological staining was used to observe the trabecular bone area fraction. Biomechanical three-point bending test was employed to measure the maximum load，stiffness，and fracture energy. Dual calcein/alizarin red fluorescence labeling and tartrate resistant acid phosphatase （TRAP） staining were applied respectively to detect the mineral apposition rate and bone formation rate as well as the number of osteoclasts on bone surfaces. Secondly，after primary osteoblasts were isolated from the tibiae of tail-suspended rats and then treated with 1 nmol/L Dex，the proportion of alkaline phosphatase （ALP）-positive osteoblasts and the activity of the enzyme were detected by ALP staining and activity test. qRT-PCR was applied to measure the expression of osteogenic activity-related factors，including osteocalcin （Ocn），Runt related transcription factor 2 （Runx2），Osterix protein （Osx），and type 1 collagen （Col1）.

‍The animal experiments revealed that Dex treatment significantly increased the tibial trabecular bone area fraction，inhibited the decrease in bone mechanical strength，and enhanced the mineralization deposition rate and new bone formation rate of trabecular bone in the tail-suspended rats （all P<0.001）. The in vitro experiments showed that Dex treatment obviously improved ALP activity and the number of ALP-positive osteoblasts in primary osteoblasts isolated from tail-suspended rats （P<0.01），and up-regulated the expression levels of osteogenic differentiation-related genes，such as Ocn，Runx2，Osx and Col1 （P<0.01）.

‍Dex exerts anti-bone loss effect in tail-suspended rats，which may be associated with its stimulation on osteoblast-mediated bone formation.

To identify the hub gene of fibroblast-like synoviocytes in rheumatoid arthritis (RA) and verify its impact on proliferation, migration and invasion of the cells.

Four independent synovial tissue microarray transcriptome sequencing datasets and one single cell sequencing dataset were downloaded from the Gene Expression Omnibus (GEO) database, and the hub gene of RA fibroblast-like synovial cells was identified by differential gene analysis, weighted gene co-expression network analysis (WGCNA) and single-cell sequencing data analysis. The expression of TNFAIP6 in human RA fibroblast-like synovial cell line MH7A was detected by RT-qPCR and Western blotting under simulated inflammatory environment. After MH7A cells were transfected with si-TNFAIP6, CCK-8 and Transwell assays were applied to detect the effects of silencing TNFAIP6 on the proliferation, migration and invasion abilities of MH7A cells.

With the aid of differential gene expression analysis, WGCNA, and single-cell sequencing data, we identified TNFAIP6 as the characteristic gene of RA fibroblast-like synoviocytes. RT-qPCR and Western blot assay demonstrated that TNFAIP6 was significantly highly expressed at mRNA and protein levels in MH7A cells stimulated with 10 ng/mL TNF-α when compared to the cells treated with PBS (P<0.05). CCK-8 and Transwell assays indicated that the silencing of TNFAIP6 markedly inhibited the proliferation, migration and invasion abilities of MH7A cells when compared with the control cells (FAM-si-NC,P<0.05).

TNFAIP6 is highly expressed specifically in RA fibroblast synoviocytes, which promotes the proliferation, migration and invasion, and may be the main contributor to the abnormal activation of RA fibroblast synoviocytes.

To explore the interaction between long non-coding RNA growth arrest-specific 5 (GAS5) and miR-135b-5p/adenomatous polyposis coli (APC) axis, and to elucidate its biological function in the proliferation, metastasis and invasion of glioma.

The relationship of GAS5 expression level with survival rate of glioma patients was analyzed based on CGGA database. qRT-PCR was used to detect the expression level of GAS5 in glioma tissues and cell lines. Human glioma T98 and A172 cell lines were subjected for overexpression and knockdown of GAS5, miR-135b-5p and APC by transfection. Western blotting and qRT-PCR were applied to measure the expression of related genes in glioma tissues or cell lines. CCK-8, Transwell and wound healing assays was conducted to determine the effect of GAS5 on the viability and motility of glioma cells. Moreover, dual luciferase reporter analysis were utilized to elucidate the regulatory mechanisms of GAS5 and miR-135b-5p/APC.

‍Analysis on CGGA database showed that the survival rate of glioma patients with low GAS5 expression was significantly decreased (P<0.001). Western blotting and qRT-PCR indicated that GAS5 was down-regulated in the glioma cell lines and tissues than human normal astrocytes and para-cancer tissues (P<0.01). The results of CCK-8, Transwell and wound healing assays revealed that overexpression of GAS5 significantly inhibited the viability, invasion and migration abilities in T98 cells when compared with the cells of the vector group (P<0.01). Dual luciferase reporter analysis displayed that there were binding sites between GAS5 and miR-135b-5p, and the expression of the two was negatively correlated (P=0.019). Further studies suggested that the effect of GAS5 on biological function in glioma cells was through targetingmiR-135b-5p, and overexpression of APC reversed the promoting effect of miR-135b-5p on glioma cells (P<0.01). Western blotting displayed that enhancing miR-135b-5p expression inhibited the expression of APC (P<0.001), and GAS5 upregulated the expression of APC by targeting miR-135b-5p (P<0.01).

GAS5 inhibits the progression of glioma via miR-135b-5p/APC axis, which revealing the molecular regulatory mechanism of GAS5/miR-135b-5p/APC.

‍To compare the efficacy of 2 L and 3 L polyethylene glycol (PEG) electrolyte solution for bowel preparation in a real-world setting.

A real-world, single-center cohort study was conducted on the individuals undergoing colonoscopy in Department of Gastroenterology of Chengdu Third People's Hospital between May and October 2023. Based on our inclusion and exclusion criteria, they were given 2 L (n=4 684) and 3 L (n=3 700) PEG electrolyte solution for bowel preparation. The primary outcome indicator was the adequacy of bowel preparation by Boston bowel preparation score (BBPS). Secondary outcome indicators included the BBPS score, polyp detection rate (PDR), tolerability, compliance, and incidence of adverse events.

‍The adequacy rate of bowel preparation was 94.35% in the 3 L group, significantly higher than that of the 2 L group (91.29%, P<0.001). The 3 L group obtained a higher BBPS score then the 2 L group (6.92±1.06 vs 6.81±1.14, P<0.001). But there was no statistical difference in the PDR between the 2 groups (P=0.073). And the rate of PEG completion (P=0.810), administration of low residue diet as required (P=0.094) or use of dimethicone (P=0.072) were comparable between the 2 groups. However, the incidences of vomiting (4.5% vs 3.2%, P=0.002), abdominal discomfort (5.0% vs 3.9%, P=0.011) and sleep disturbance (18.0% vs 14.6%, P<0.001) were obviously higher in the 3 L group than the 2 L group.

In a real-world setting, 2 L PEG is a considerably safe and effective regimen for bowel preparation.

‍To investigate the role and mechanism of bergapten in treating osteoarthritis (OA) based on network pharmacology, molecular docking and in vitro experiments.

‍The functional targets of bergapten and the related targets of OA disease were obtained by searching Super-Pred, Swiss Target Prediction, Drug Bank. The obtained targets were intersected with Venny2.1.0, protein-protein interaction (PPI) analysis was performed using the STRING database, and molecular docking was conducted by AutoDock Vina. Subsequently, human chondrocyte C28/I2 cells were subjected in the following validation experiments. After the cells were treated with 0~50 μmol/L bergapten for 24, 48 or 72 h, CCK-8 assay, RT-qPCR and immunofluorescence assay were used to detect the proliferation and apoptosis, and the expression of the molecules related to proliferation, inflammation and senescence (senescence associated secretory phenotype, SASP). Senescence β-galactosidase (SA-β-Gal) staining kit was employed to detect the change of SA-β-Gal level in the cells.

There obtained 145 potential targets of bergapten for OA. PPI analysis revealed 4 potential core targets, HSP90AA1, EGFR, HIF1A, and PIK3CA, and they could form relatively stable complex with bergapten. CCK-8 assay showed that 10 μmol/L bergapten treatment for 48 h resulted in the highest proliferative activity of C28/I2 cells (P<0.05). Immunofluorescence assay, RT-qPCR and Western blotting indicated that bergapten treatment induced significant increases in the expression of cell proliferation markers such as PCNA, CCND1 and CCNE1 (P<0.05), while decreases in the expression of apoptosis markers Caspase3 and BAX (P<0.05). Meanwhile, the expression levels of IL-1β-induced inflammatory factors IL-6 and TNF-α were reduced, while those of anti-inflammatory factors IL-4 and IL-10 were elevated after bergapten treatment (P<0.05). The levels of SASP factors, such as P16, P21, MMP9 and MMP13 were also significantly declined (P<0.05). SA-β-Gal staining displayed that the level of SA-β-Gal in C28/I2 cells was significantly enhanced after IL-1β induction (P<0.05), while bergapten treatment could decrease the positive cell rate of the staining (P<0.05).

Bergapten can reduce the expression of SASP factors such as IL-6, TNF-α, P16, P21, MMP9 and MMP13 in IL-1β-stimulated chondrocytes, and thus exerts its anti-inflammatory and anti-aging effects in delaying the progression of OA.

‍To investigate the prognostic factors affecting survival in patients with natural killer T-cell lymphoma (NKTL), and then develop a prognostic model for predicting their overall survival (OS) based on random survival forest (RSF) algorithm.

Demographic and clinical pathological data of NKTL patients were collected from the SEER database during 2000 and 2020. The patients were divided into a training cohort (n=471) and a validation cohort (n=203) in a 7∶3 ratio. Cox regression analysis was performed to identify prognostic factors affecting OS, and a nomogram model was constructed based on the obtained factors. Meanwhile, RSF algorithm was used to determine prognostic factors affecting OS to build the RSF model. The models were evaluated using receiver operating characteristic (ROC) curve, calibration curve, decision curve, net reclassification improvement (NRI), and integrated discrimination improvement (IDI), and the predictive performances of the 2 models were compared. Risk scores for each patient were calculated using the 2 models. Then the patients were divided into high- and low-risk groups based on the median risk score, and survival curve was plotted for comparison.

Ann Arbor stage, age, radiotherapy, combined treatment, and type of disease were identified as significant prognostic variables associated with OS. In the validation cohort, the area under the ROC curve (AUC) for the nomogram model at 1, 3, and 5 years was 0.745, 0.771, and 0.748, respectively, while the AUC for the RSF model was 0.764, 0.792, and 0.761 at the same time points. ROC curve analysis indicated that both models demonstrated good accuracy and discrimination in predicting OS. Calibration curve analysis showed a strong consistency between the predicted and actual OS for both models. Both models effectively stratified the patients into poor and favorable prognosis groups, with the OS of patients in the poor prognosis group being significantly shorter than that of the favorable prognosis group (P<0.000 1). Decision curve analysis revealed that the net benefit of the RSF model was superior to that of the nomogram model. Compared to the nomogram model, the NRI for the RSF model was 0.184 (95%CI: 0.098~0.267, P<0.01), and the IDI was 0.300 (95%CI: 0.241~0.359, P<0.01). Overall, the RSF model demonstrated superior predictive capability than the nomogram model.

Ann Arbor stage, age, radiotherapy, combined treatment, and type of disease are prognostic factors affecting the prognosis of NKTL patients. Our RSF model demonstrates strong predictive capability for the prognosis of NKTL patients and can effectively assess patient outcomes.