Injury of vascular endothelial barrier function is implicated in several pathophysiological processes. The integrity of vascular endothelium is regulated by cytoskeleton and cell-cell junctions. Small guanosine triphosphatases of the Rho family (Rho GTPases) are known to play a central role in vascular endothelial barrier function. It has been reported that RhoA, Rac1, Cdc42 and RhoB are involved and they exert both positive and negative effect on endothelial barrier integrity, depending on their subcellular location. When inflammatory factors such as thrombin attack the vascular endothelial cells, GEF of RhoA will be widely distributed throughout the cells. Thus, activated RhoA causes aggregation of F-actin fibers in a short time and disrupts the vascular endothelial barrier, a process named acute cell contraction. However, RhoA may also induce the production and maturation of intercellular junctions in new cells. Rac1 and Cdc42 help to maintain the integrity of vascular endothelial barrier at the resting state. They cause the phosphorylation of LIM kinase and inhabitation of cofilin, resulting in less remodeling of cytoskeletal in the vascular endothelial cells. On the other hand, Cdc42 can translocate to the cortex rapidly after a stimulation, where Cdc42 will activate the myosin Ⅱ and promote the reorganization of adjective junction to facilitate the recovery of vascular endothelial barrier. In this review, we overviewed how Rho GTPases regulate the vascular endothelial barrier integrity.

In ischemic stroke, increased level of neuronal complex of nitric oxide synthase (nNOS)-postsynaptic density protein-95 (PSD-95) plays an important role in neuronal damage. We aimed to establish a screening model to identify compounds capable of uncoupling nNOS interaction with PSD-95. In this model, human embryonic kidney-293T (HEK-293T) cells were transfected with either pCDH-Flag-nNOS or pcDNA3.1-PSD-95 plasmid to obtain the protein of Flag-nNOS or PSD-95. Incubating Flag-nNOS with PSD-95 causes formation of the nNOS-PSD-95 complex. ZL006, a known uncoupler of nNOS-PSD-95 interaction, can disturb the interaction between Flag-nNOS and PSD-95, serving as a positive control. The method coupling antibodies to magnetic beads with glutaraldehyde was used to decrease the cost and increase the efficiency. To establish that our model is suitable for selecting nNOS-PSD-95 uncouplers, we evaluated the ability of IC87201, another reported uncoupler of nNOS-PSD-95 interaction, and structural analogs of ZL006. IC87201 and one structure analog of ZL006 showed uncoupling effect, supporting that our model can be used to select different types uncoupler blocking nNOS-PSD-95 interaction.

Danhong injection (DHI) and ceftriaxone sodium were used in combination based on their experimental uses in clinic. This study was designed to investigate the impact of ceftriaxone on pharmacokinetics and pharmacodynamics of the phenolic acids from DHI. After administration of DHI for 7 d, ceftriaxone (CFTX) was combined with DHI for the next 7 d in adult male Sprague-Dawley (SD) rats. All the drugs were administered through caudal vein. UHPLC-TQ-MS was applied in determining the plasma concentration of p-coumaric acid (p-CA), salvianolic acid D (SaD), rosmarinic acid (RA) and salvianolic acid B (SaB). The pharmacokinetic parameters of the combination group or the Danhong injection alone group were calculated by statistical moment method, C_max and the average of the area under the curve AUC_0-t using 90% confidence interval of the bioequivalence and bioavailability degree module in DAS 3.2.8 statistic software. The results showed that C_max of p-CA, SaD, RA and SaB were unqualified within 90% confidence intervals for bioequivalence statistics. And the results showed that AUC_0-t of SaD, RA and SaB within 90% confidence intervals for bioequivalence statistics were unqualified. There were no significant difference in the t_max (P>0.05). The results of anticoagulation in vivo showed that the international normalized ratio (INR), prothrombin time (PT), thrombin time (TT) and activated partial thromboplastin time (APTT) were significantly increased when combined with CFTX (P < 0.05 or P < 0.01). The results in antithrombotic effects revealed that the thromboxane B₂ (TXB₂) level in serum was significantly decreased (P < 0.01) in the combination group compared with Danhong injection alone. However, there was no significant difference in antiplatelet effects. These results suggest that CFTX may enhance the anticoagulation and antithrombotic effects of DHI through altering pharmacokinetics and pharmacodynamics in SD rats.

This study aimed to investigate the effect of Huangqin Tang (HQT) on oxidative stress associated with ulcerative colitis in rats, and to explore its antioxidant mechanism. After approved by Institute of Chinese Materia Medica Ethics Committees in China Academy of Chinese Medical Sciences, the rats were given 2, 4, 6-trinitrobenzenesulfonic (TNBS)/ethanol mixture to induce the ulcerative colitis (UC), and were randomly divided into normal group, model group, the salazosulfapyridine (SASP) group, and high, middle or low dose (20, 10, 5 g·kg^-1) of HQT groups. After 5 days of treatment, the activity of catalase (CAT) from micrococcus lysodeikticus, glutathione peroxidase (GSH-px), myeloperoxidase (MPO), superoxide dismutase (SOD) were detected by biochemical assays. The levels of lipid peroxide (LPO) were detected by ELISA. The positive protein expression of nuclear factor erythroid-2-related factor 2 (Nrf2) were detected by immunohistochemistry method and the downstream antioxidant enzymes of Nrf2 were determined by Western blot analyses. The levels of SOD, CAT and GSH-px activities in the normal group were significantly higher than the model group, while the serum MPO activity in the model group was obviously increased (P < 0.05 or P < 0.01). Compared with the model group, there was a significant difference in the activity of CAT in the high and middle dose groups of HQT (P < 0.05 or P < 0.01), the activity of GSH-px in the high, middle and low dose groups of HQT were apparently higher than the model group (P < 0.05); The serum levels of LPO in the model group were significantly higher than those in the normal group (P < 0.05), while the up-regulating effects on LPO were reversed by the high and middle dose groups of HQT (P < 0.05). The expression of Nrf2 in the high-dose group of HQT and SASP group was statistically significant (P < 0.05). The results of Western blot showed that compared with the model group, each of the HQT and SASP group could increase the heme oxygenase (HO-1) and NAD[P]H:quinone oxidoreductase 1 (NQO-1) expression in a dose-dependence manner. HQT has significant anti-oxidative stress and obviously improves the signs, mental status and defecation of UC rats. The mechanism of action for HQT maybe related to activate the Nrf2 pathway and increase the expression of Ⅱ phase metabolic enzymes such as HO-1 and NQO-1, reduce the content of LPO and MPO in serum and enhance the activity of SOD, CAT and GSH-px.

This study explores the antidepressant mechanism of Radix Bupleuri-Radix Paeoniae Alba herb pair and the core role in Xiaoyaosan. The behavioral indicators were used to observe the effects of herb pair, Xiaoyaosan, and a negative control (Xiaoyaosan remove Radix Bupleuri-Radix Paeoniae Alba) on CUMS rats. The body weight, number of crossing and rearing, sugar preference rate were significantly decreased (P < 0.001) and the immobility time of forced swimming test was significantly prolonged (P < 0.01) in the model group. Radix Bupleuri-Radix Paeoniae Alba herb pair has significant antidepressant effect, with the number of crossing in low dose group similar to Xiaoyaosan, while the number of rearing and the sucrose preference rate in high dose group are equivalent to Xiaoyaosan, and the immobility time in high and low-doses group are better than Xiaoyaosan. Animal experimentation was approved according to the Committee on the Ethics of Animal Experiments of Shanxi University. Metabonomics results showed 15 biomarkers related to depression, whereas administering Xiaoyaosan or the herb pair can ameliorate 12 or 8 metabolites respectively, compared to the negative control, which can only ameliorate 6 metabolites. The metabolic pathway analysis showed 5 depression-related pathways. The herb pair and Xiaoyaosan exerted antidepressant effects through common 4 metabolic pathways, while the negative control only exerted effects through 3. In summary, behavioral, metabolomics and metabolic pathway analysis revealed that Radix Bupleuri-Radix Paeoniae Alba plays an important role in the biological effect of Xiaoyaosan.

Ellagic acid is ubiquitous in plants and is considered as a potential candidate for antioxidant and antineoplastic drugs. However, ellagic acid has poor solubility and precipitates easily even after initial solubilization. Improvement of its bioavailability has been a concern of pharmaceutical industry. It was found that storage in Sanlejiang oral liquid at low temperature keeps its stability. Ellagic acid is anomalous in a way that is easily soluble at low temperatures but precipitates at high temperatures. In order to reveal the mechanism of this phenomenon and develop precipitation prevention and control strategies, ellagic acid in Sanlejiang oral liquid was stored at high, medium and low temperatures for three months. The changes of composition and phase state of the whole system during storage were systematically tracked and studied by means of precipitation amount or morphology, HPLC chemical profile of supernatant versus precipitates, and comprehensive characterization of physical phase state. The results show that the amount of precipitation at low temperature is only 1/3 of that at normal room temperature. As the temperature rises, the sedimentation increases sharply. HPLC analyses of supernatant versus precipitates revealed that ellagic acid precipitation originated from two ways:chemical degradation and physical deposition. The chemical sedimentation is related to the hydrolysis of tannins under acidic condition, forming chebulagic acid and corilagin. Physical sedimentation is related to the decrease of the association degree and viscosity of polyphenol colloids when temperature rises. This study elucidated the stability mechanism of ellagic acid in liquid preparations of TCM, and provided the mechanistic basis for efficient utilization and solution prepartion of ellagic acid.

In this study, black phosphorus quantum dots (BPQDs)-loaded liposomes (liposome-BPQDs) were prepared to explore physicochemical properties and photothermal effects on cervical cancer cells. BPQDs were fabricated by ultrasonic method. Liposome-BPQDs were prepared by thin film dispersion. Surface morphology, particle size, zeta potential and Raman spectra of liposome-BPQDs were characterized. The cytotoxicity of the liposome-BPQDs against human cervical cancer cells (HeLa) was examined by CCK-8 assay. Confocal laser scanning microscope (CLSM) and fluorescence microscopy were used to observe the uptake and apoptosis of HeLa cells. The results indicated that liposome-BPQDs were ellipsoidal or spherical under scanning electron microscope, TEM observation showed liposome-BPQDs were about 90-110 nm in diameter. The particle size measurements showed liposome-BPQDs were (104.2±0.35) nm in diameter, and zeta potential were examined to be (-11.3±3.01) mV. The encapsulation efficiency was (84.40±2.13)%.Under natural conditions with outdoor ventilation, temperature range of 25℃-34℃ and relative humidity of 80%-82%, the photothermal effects of liposome-BPQDs was better and the degradation denaturation of liposome-BPQDs were slower than those of BPQDs. The results also reflected that liposome-BPQDs could be uptaken by HeLa cells easily. After near-infrared laser irradiation, the mortality of HeLa cells rise significantly when the amount of BPQDs reach 20 μg·mL^-1. In summary, liposome-BPQDs with high stability exhibited good photothermal effects, which can be expected to be applied to photothermal therapy of cervical carcinoma.

The chemical constituents of Litsea coreana were investigated using chromatographic methods, including column chromatography on silica gel, MCI, and semi-preparation HPLC. Two compounds were isolated and identified as hawktealignan A (1) and cinnamophilin (2) by NMR analyses as well as their physical and chemical properties. Compound 1 is a new lignan and compound 2 was isolated from this plant for the first time.

In this paper, three methods, differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and X-ray powder diffraction (PXRD), were used to characterize the structure of Palbociclib. The form-B crystal mixed in with form-A crystal, the effective form of Palbociclib, was quantitated by the single peak method (peak area method and peak height method) using X-ray powder diffraction. While the qualitative identification by DSC was not clear, SEM and PXRD quickly and effectively identified two types of crystal. The standard curve equations established by the peak area method and the peak height method are:y=0.842 75x-0.001 21 and y=0.909 64x-0.002 32. This suggests that the linear relationship of peak area method is better than that of peak height method (R²=0.986 17 and R²=0.985 83). The sensitivity of the peak area method (LOD=1.17%, LOQ=3.92%) are higher than the peak height method (LOD=1.19%, LOQ=3.97%). The methods from this study can be used to identify and quantify palbociclib form-A and form-B crystal rapidly.

Chemotherapy plays an essential role in controlling tumor growth and progression. However, long-term use of chemotherapeutic drugs usually results in drug resistance in tumor cells, leading to treatment failure and disease progression. The mechanism of tumor resistance to chemotherapy and the strategy of prevention or reversal of such resistance have always been hot issues in cancer therapy research. Exosomes are small spherical vesicles secreted by cells with a diameter of 40-100 nm. They carry a variety of bioactive small molecules (including DNA, ncRNA, RNA, and proteins) and participate in regulation of cell microenvironment, thereby affecting a variety of physiological and pathological activities in the body. In recent years, studies have shown that exosomes play an important role in cancer cell resistance to chemotherapy, metastasis, and immune escape. This article reviews the role and mechanism of exosomes in the development of drug resistance in tumors, and aims to provide new ideas for the prevention or treatment of tumor resistance.