Mechanism of levosimendan in treating hypoxic pulmonary hypertension based on network pharmacology and molecular docking technology / 中国药理学通报

Aim To explore the efficacy of levosimendan on hypoxia pulmonary hypertension through animal experiments, and to further explore the potential mechanism of action using network pharmacological methods and molecular docking technique. Methods The rat model of hypoxia pulmonary hypertension was constructed to detect right heart systolic pressure and right heart remodeling index. HE , Masson, and VG staining were core targets were screened out. GO and KEGG pathway enrichment analysis were performed using the DAVID database. Molecular docking of the core targets was performed with the AutoDock software. Results The results of animal experiments showed that levosimendan had obvious therapeutic effect on hypoxia pulmonary hypertension. The network pharmacology results showed that SRC, HSP90AA1, MAPK1, PIK3R1, AKT1, HRAS, MAPK14, LCK, EGFR and ESR1 used to analyze the changes of rat lung histopathology. Search the Swiss Target Prediction, DrugBank Online, BatMan, Targetnet, SEA, and PharmMapper databases were used to screen for drug targets. Disease targets were retrieved from the GeneCards, OMIM databases. The "drug-target-disease" network was constructed after identification of the two intersection targets. The protein interaction network was constructed and the were the key targets to play a therapeutic role. Molecular docking showed good docking of levosimendan with all the top five core targets with degree values. Conclusions Levosimendan may exert a therapeutic effect on hypoxia-induced pulmonary hypertension through multiple targets.

Hydroxysafflor Yellow A Promotes HaCaT Cell Proliferation and Migration by Regulating HBEGF/EGFR and PI3K/AKT Pathways and Circ_0084443 / 中国结合医学杂志

OBJECTIVE@#To investigate the effect and possible mechanism of hydroxysafflor yellow A (HSYA) on human immortalized keratinocyte cell proliferation and migration.@*METHODS@#HaCaT cells were treated with HSYA. Cell proliferation was detected by the cell counting kit-8 assay, and cell migration was measured using wound healing assay and Transwell migration assay. The mRNA and protein expression levels of heparin-binding epidermal growth factor (EGF)-like growth factor (HBEGF), EGF receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR), and hypoxia-inducible factor-1α (HIF-1α) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Circ_0084443-overexpressing HaCaT cells and empty plasmid HaCaT cells were constructed using the lentiviral stable transfection and treated with HSYA. The expression of circ_0084443 was detected by qRT-PCR.@*RESULTS@#HSYA (800 µmol/L) significantly promoted HaCaT cell proliferation and migration (P<0.05 or P<0.01). It also increased the mRNA and protein expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and increased the phosphorylation levels of PI3K and AKT (P<0.05 or P<0.01). Furthermore, HSYA promoted HaCaT cell proliferation and migration via the HBEGF/EGFR and PI3K/AKT/mTOR signaling pathways (P<0.01). Circ_0084443 attenuated the mRNA expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α (P<0.05). HSYA inhibited the circ_0084443 expression, further antagonized the inhibition of circ_0084443 on HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and promoted the proliferation of circ_0084443-overexpressing HaCaT cells (P<0.05 or P<0.01). However, HSYA could not influence the inhibitory effect of circ_0084443 on HaCaT cell migration (P>0.05).@*CONCLUSION@#HSYA played an accelerative role in HaCaT cell proliferation and migration, which may be attributable to activating HBEGF/EGFR and PI3K/AKT signaling pathways, and had a particular inhibitory effect on the keratinocyte negative regulator circ_0084443.

Mechanism of albiflorin in improvement of Alzheimer's disease based on network pharmacology and in vitro experiments / 中国中药杂志

This study aimed to explore the mechanism of albiflorin in the treatment of Alzheimer's disease(AD) based on network pharmacology, molecular docking, and in vitro experiments. Network pharmacology was used to predict the potential targets and pathways of albiflorin against AD, and molecular docking technology was used to verify the binding affinity of albiflorin to key target proteins. Finally, the AD cell model was induced by Aβ_(25-35) in rat pheochromocytoma(PC12) cells and intervened by albiflorin to validate core targets and pathways. The results of network pharmacological analysis showed that albiflorin acted on key targets such as mitogen-activated protein kinase-1(MAPK1 or ERK2), albumin(ALB), epidermal growth factor receptor(EGFR), caspase-3(CASP3), and sodium-dependent serotonin transporter(SLC6A4), and signaling pathways such as MAPK, cAMP, and cGMP-PKG. The results of molecular docking showed that albiflorin had strong binding affinity to MAPK1(ERK2). In vitro experiments showed that compared with the blank group, the model group showed decreased cell viability, decreased expression level of B-cell lymphoma 2(Bcl-2), increased Bcl-2-associated X protein(Bax), and reduced phosphorylation level of extracellular signal-regulated kinase 1/2(ERK1/2) and the relative expression ratio of p-ERK1/2 to ERK1/2. Compared with the model group, the albiflorin group showed potentiated cell viability, up-regulated expression of Bcl-2, down-regulated Bax, and increased phosphorylation level of ERK1/2 and the relative expression ratio of p-ERK1/2 to ERK1/2. These results suggest that the mechanism of albiflorin against AD may be related to its activation of the MAPK/ERK signaling pathway and its inhibition of neuronal apoptosis.

Serum metabolomics study of Psoraleae Fructus in improving learning and memory ability of APP/PS1 mice / 中国中药杂志

This study aimed to investigate the mechanism of Psoraleae Fructus in improving the learning and memory ability of APP/PS1 mice by serum metabolomics, screen the differential metabolites of Psoraleae Fructus on APP/PS1 mice, and reveal its influence on the metabolic pathway of APP/PS1 mice. Thirty 3-month-old APP/PS1 mice were randomly divided into a model group and a Psoraleae Fructus extract group, and another 15 C57BL/6 mice of the same age were assigned to the blank group. The learning and memory ability of mice was evaluated by the Morris water maze and novel object recognition tests, and metabolomics was used to analyze the metabolites in mouse serum. The results of the Morris water maze test showed that Psoraleae Fructus shortened the escape latency of APP/PS1 mice(P<0.01), and increased the number of platform crossing and residence time in the target quadrant(P<0.01). The results of the novel object recognition test showed that Psoraleae Fructus could improve the novel object recognition index of APP/PS1 mice(P<0.01). Eighteen differential metabolites in serum were screened out by metabolomics, among which the levels of arachidonic acid, tryptophan, and glycerophospholipid decreased after drug administration, while the levels of glutamyltyrosine increased after drug administration. The metabolic pathways involved included arachidonic acid metabolism, glycerophospholipid metabolism, tryptophan metabolism, linoleic acid metabolism, α-linolenic acid metabolism, and glycerolipid metabolism. Therefore, Psoraleae Fructus can improve the learning and memory ability of APP/PS1 mice, and its mechanism may be related to the effects in promoting energy metabolism, reducing oxidative damage, protecting central nervous system, reducing neuroinflammation, and reducing Aβ deposition. This study is expected to provide references for Psoraleae Fructus in the treatment of Alzheimer's disease(AD) and further explain the mechanism of Psoraleae Fructus in the treatment of AD.

Comparison of efficacy of arthroscopic all-inside ligament repair with suture augmentation and ligament reconstruction with tendons in the treatment of chronic ankle instability / 中华创伤杂志

Objective:To compare the efficacy of arthroscopic all-inside ligament repair with suture augmentation and ligament reconstruction with tendons in the treatment of chronic ankle instability with poor remnant quality of the anterior talofibular ligament (ATFL) tissue.Methods:A retrospective cohort study was conducted to analyze the clinical data of 37 patients with chronic ankle instability treated at Huashan Hospital Affiliated to Fudan University from January 2018 to August 2020, including 34 males (34 ankles) and 3 females (3 ankles); aged 18-57 years [(32.2±7.2)years]. The time from injury to operation ranged from 3-360 months [48(12, 120)months]. All patients underwent arthroscopic all-inside ankle stabilization surgery, of which 19 underwent ligament repair with suture augmentation (augmented repair group) and 18 underwent traditional ligament reconstruction with allograft/autograft tendons (tendon reconstruction group). The degree of ATFL injury and intra-articular lesions (osteophytes, loose bodies and cartilage damage) were recorded during the operation. The Karlsson scale and Tegner scale were evaluated before operation and at the last follow-up. The number of patients who were able to complete partial weight-bearing/return to normal walking/return to sports postoperatively and the time required were compared between the two groups. Postoperative complications were observed.Results:All patients were followed up for 12-32 months [21(16, 28)months]. There were no significant differences in the degree of ATFL injury and intra-articular lesions (osteophytes, loose bodies and cartilage damage) seen during the operation between the two groups (all P>0.05). At the last follow-up, the Karlsson score in augmented repair group and tendon reconstruction group [95.0(90.0, 98.5)points and 95.0(87.8, 99.3)points] was significantly higher than the preoperative level [65.0(51.0, 75.0)points and 65.0(53.3, 78.0)points] (all P<0.01). At the last follow-up, the Tegner score in augmented repair group and tendon reconstruction group [5.0(3.5, 6.0)points and 5.0(3.3, 6.0)points] were significantly higher than the preoperative level [3.0(2.0, 4.0)points and 2.5(1.3, 4.0)points] (all P<0.01). There were no significant differences in Karlsson score and Tegner score between the two groups (all P>0.05). All patients completed partial weight-bearing after 3.0(2.0, 4.0)weeks in augmented repair group and 4.0(3.5, 6.0)weeks in tendon reconstruction group. All patients returned to normal walking after 8.0(6.0, 9.0)weeks in augmented repair group and 8.0(5.5, 12.0)weeks in tendon reconstruction group. A total of 13 patients (63%) in augmented repair group and 13 patients (72%) in tendon reconstruction group successfully returned to sports postoperatively and the time required was 6.0(3.5, 8.0) months and 6.0(4.5, 12.0)months, respectively. There were no significant differences in the above indicators between the two groups (all P>0.05), but augmented repair group had a trend of faster completion of partial weight-bearing than tendon reconstruction group. There was 1 patient [5%(1/19)] in augmented repair group and 1 patient [6%(1/18)] in tendon reconstruction group who reported feelings of instability during exercise postoperatively ( P>0.05). None of the patients in augmented repair group had limited ankle range-of-motion, not different from 1 patient [6%(1/18)] in tendon reconstruction group ( P>0.05). Conclusion:In the treatment of chronic ankle instability with poor remnant quality of the anterior talofibular ligament (ATFL) tissue, both arthroscopic all-inside ligament repair with suture augmentation and ligament reconstruction with tendons can improve the short-term postoperative ankle function and activity level of the patients, and the former one has advantages such as simple operative procedures and none use of grafts.

in vitro metabolism of daphnetin in rat liver S9 fractions / 药学学报

Daphnetin is quickly eliminated in rats after dosing, but the mechanism remains unclear. This study was aimed to investigate the in vitro metabolism of daphnetin using rat liver S9 fractions (RLS9). The metabolites formed in RLS9 were identified and the kinetic parameters for different metabolic pathways were determined. HPLC-DAD-MS analysis showed that daphnetin was biotransformed to six metabolites, which were identified as 7 or 8 mono-glucuronide and mono-sulfate, 8-methylate, and 7-suflo-8-methylate. Methylation and glucuronidation of daphnetin exhibited the Michaelis-Menten kinetic characteristics, whereas the substrate inhibition kinetic and the two-site kinetic were observed for 8-sulfate and 7-sulfate formations. Of the 3 conjugation pathways, the intrinsic clearance rate for sulfation was highest, followed by methylation and glucuronidation. By in vitro-in vivo extrapolation of the kinetic data measured in RLS9, the hepatic clearance were estimated to be 54.9 mL·min-1·kg-1 which is comparable to the system clearance (58.5 mL·min-1·kg-1) observed in rats. In conclusions, the liver might be the main site for daphnetin metabolism in rats. Sulfation, methylation and glucuronidation are important pathways of the hepatic metabolism of daphnetin in rats.

The clinical evaluation of the effect of two different apical surgery timing / 实用口腔医学杂志

Objective:To observe the effect of two different apical surgery timing. Methods:68 patients with periapical lesion were divided into 2 groups. 30 patients( control group) were operated by apical surgery at least 1-2 months after root canal therapy( RCT) , while 38 patients(experimental group) were operated immediately after RCT. The patients were followed up 3, 6 and 12 months after apical surgery. Results:The curative effect analysed with the age, sex and tooth position showed no statistical difference between 2 groups(P>0. 05). Conclusion:Immediate and delayed apical surgery apical surger after RCT are similarly effective.