[Objective]To investigate the mechanism of Compound Fuling Granule(CFG)in inhibiting the glucose metabolism and metastasis of ovarian cancer cells.[Methods]Ovarian cancer cell HEY-T30 were cultured in vitro and incubated with different concentration(0.25,0.5,1,1.5,2 mg·mL-1)of CFG for 24 h.Cell counting kit-8(CCK-8)assay was performed to detect the effect of CFG on the cell proliferation,Transwell assay was used to investigate cell migration ability,lactic acid detection kit and glucose detection kit were used to detect lactic acid production and glucose consumption,the expressions of dynamin-related protein 1(DRP1),some key glycolysis-related proteins and metastasis-related proteins were detected by Western blot,Mito-Tracker Red was used to label mitochondria to observe mitochondria morphology.Lentivirus transfection technique was used to achieve the stable DRP1 knockdown HEY-T30 cells.Real-time quantitative polymerase chain reaction(Real-time qPCR)was used to detect the expression of DRP1 mRNA,the effect of CFG on lactic acid production and glucose consumption of HEY-T30 after DRP1 knockdown was detected by lactic acid detection kit and glucose detection kit,Transwell assay was used to investigate the migration ability of HEY-T30 with DRP1 knockdown after treated with CFG,the effect of CFG on the expression of glycolysis-related proteins and metastasis-related proteins in HEY-T30 with DRP1 knockdown was detected by Western blot.[Results]Compared with control group,the cell survival rate in 0.5,1,1.5,2 mg·mL-1 CFG groups reduced significantly(P＜0.01).The average length of mitochondria in 0.5,1 mg·mL-1 group was markedly increased(P＜0.01),the protein expression of DRP1 was significantly reduced(P＜0.05,P＜0.01),and the lactate production and glucose consumption in 0.5 and 1 mg·mL-1 groups were significantly decrease(P＜0.01).The number of migration cell was significantly reduced in 0.25,0.5 and 1 mg·mL-1 concentration groups(P＜0.05,P＜0.01).After knockdown of DRP1,the glycolysis level and migration of HEY-T30 were decreased(P＜0.05,P＜0.01),and the expressions of glycolysis-related proteins and metastasis-related proteins were decreased(P＜0.05,P＜0.01).The inhibitory effect of CFG on glycolysis and metastasis of ovarian cancer cells was also weakened.[Conclusion]By targeting DRP1 to regulate glucose metabolism reprogramming,CFG could inhibit the metastasis of ovarian cancer.

Objective:To evaluate the role of ubiquitin-specific peptidase 22 (USP22) in myocardial ischemia-reperfusion (I/R) injury in diabetic mice.Methods:Seventy-eight SPF male C57BL/6 mice, aged 6-8 weeks, were divided into 6 groups using a random number table method: sham operation group (Sham group, n=12), type 1 diabetes mellitus + sham operation group (T1D+ Sham group, n=12), myocardial I/R injury group (I/R group, n=12), type 1 diabetes mellitus + myocardial I/R injury group (DI/R group, n=12), type 1 diabetes mellitus + myocardial I/R injury + empty vector group (DI/R+ V group, n=15), and type 1 diabetes mellitus + myocardial I/R injury + USP22 overexpression group (DI/R+ U group, n=15). Type 1 diabetes mellitus was induced by intraperitoneal injection of streptozotocin-citrate buffer. Myocardial I/R was induced by ligation of the left coronary artery. At 1 day before developing the myocardial I/R injury model, DI/R+ U group and DI/R+ V group received an intramyocardial injection of USP22 overexpression plasmid or empty vector plasmid, respectively. At 24 h of reperfusion, cardiac function was assessed using the echocardiography to measure the left ventricular ejection fraction and left ventricular fractional shortening. The mice were then sacrificed, and their hearts were harvested for measurement of the myocardial infarct size, for microscopic examination of pathological changes (using HE staining) and for determination of the apoptosis rate (TUNEL staining), reactive oxygen species(ROS) activity (DHE staining), and USP22 expression (by Western blot, immunofluorescence, and immunohistochemistry). Proteomic analysis was performed to identify downstream proteins regulated by USP22, and protein-protein interactions were investigated using co-immunoprecipitation. Results:Compared with Sham group, the cardiac function indices were significantly decreased, the apoptosis rate of myocardial cells and ROS activity were increased, and USP22 expression in myocardial tissues was down-regulated in I/R group ( P<0.05). Compared with I/R group, the percentage of myocardial infarct size was significantly increased, the cardiac function indices were decreased, the apoptosis rate of myocardial cells and ROS activity were increased, and USP22 expression in myocardial tissues was up-regulated ( P<0.05), and the pathological damage to myocardial tissues was aggravated in DI/R group. Compared with DI/R+ V group, the percentage of myocardial infarct size was significantly decreased, the cardiac function indices were increased, the apoptosis rate of myocardial cells and ROS activity were decreased, and USP22 expression in myocardial tissues was up-regulated ( P<0.05), and the pathological damage to myocardial tissues was alleviated in DI/R+ U group. The results of proteomics combined with co-immunoprecipitation experiments showed an interaction between calponin 1 and USP22. Conclusions:During myocardial I/R injury in diabetic mice, USP22 may act as an endogenous protective mechanism, and calponin 1 might be a downstream mechanism through which USP22 exerts its protective effects.

OBJECTIVE： To study the anti-inflammatory effect and its mechanism of the extract of Dcsmodium microphyllum， so as to provide experiment reference for further study of D. microphyllum. METHODS： Acute inflammatory model was established by xylene，glacial acetic acid and carrageenan. Using dexamethasone as positive control （0.005 g/kg）， inhibitory effects of intragastric different doses of the extract of D. microphyllum （50， 30， 15 g/kg） on xylene-induced ear swelling in normal mice and adrenalectomized mice， glacial acetic acid-induced permeability increasing of abdominal capillaries in normal mice， carrageenan-   induced paw swelling in normal mice and adrenalectomized mice were investigated. The levels of MDA， SOD and NO in the inflammatory tissue of toes of adrenalectomized mice were detected in carrageenan-induced inflammation model. Blank group was set for control （ig. equal volumn of water）. RESULTS： Compared with blank group， ear swelling degree of normal mice and adrenalectomized mice were decreased significantly in D. microphyllum extract high-dose and medium-dose groups while inhibitory rate of ear swelling was increased significantly； the permeability of abdominal capillaries of normal mice was significantly decreased in D. microphyllum extract groups； the swelling degree of toes in normal mice of D. microphyllum extract high-dose and middle-dose groups and adrenalectomized mice were significantly decreased while inhibitory rate of toe swelling was increased significantly （P＜0.05 or P＜0.01）. The levels of MDA and NO in the toe inflammatory site of adrenalectomized mice were decreased significantly in D. microphyllum extract high-dose and medium-dose groups， while the level of SOD was increased significantly （P＜0.05）. CONCLUSIONS： D. microphyllum extract can inhibit acute inflammation in mice significantly. Its anti-inflammatory mechanism is associated with decreasing MDA and NO while increasing SOD levels， and the anti-inflammatory effect does not depend on the hypothalamus-pituitary-adrenal axis system.