Exploration of anti­osteosarcoma activity of asiatic acid based on network pharmacology and in vitro experiments.

Osteosarcomas are malignant bone tumors that typically originate in the epiphyses of the long bones of the extremities in adolescents. Asiatic acid has been reported to possess anti­inflammatory, neuroprotective, antidiabetic, antitumor and antimicrobial activities. The present study used a combination of network pharmacological prediction and in vitro experimental validation to explore the potential pharmacological mechanism of asiatic acid against osteosarcoma. A total of 78 potential asiatic acid targets in osteosarcoma were identified using databases. Kyoto Encyclopedia of Genes and Genomes analysis indicated that the PI3K/AKT and MAPK signaling pathways are essential in the treatment of osteosarcoma with asiatic acid. Molecular docking revealed binding of asiatic acid to EGFR, Caspase­3, ESR1, HSP90AA1, IL­6 and SRC proteins. asiatic acid inhibited proliferation through G2/M cell cycle arrest in osteosarcoma cells. In addition, asiatic acid induced mitochondria­dependent apoptosis as demonstrated by increases in Bax and VDAC1 expression, and a decrease in Bcl­2 protein expression. The increased autophagosomes, increased LC3­II/I ratios and decreased p62 expression in the treatment group indicated that asiatic acid triggered autophagy. In addition, asiatic acid decreased the levels of phosphorylated (p­)PI3K/PI3K and p­AKT/AKT, increased reactive oxygen species (ROS) and upregulated the levels of p­ERK1/2/ERK1/2, p­p38/p38 and p­JNK/JNK in osteosarcoma cells. These results demonstrated that asiatic acid inhibited osteosarcoma cells proliferation by inhibiting PI3K/AKT and activating ROS/MAPK signaling pathways, suggesting asiatic acid is a potential agent against osteosarcoma.

CCL2 promotes proliferation, migration and angiogenesis through the MAPK/ERK1/2/MMP9, PI3K/AKT, Wnt/ß­catenin signaling pathways in HUVECs.

Severe bone trauma can lead to poor or delayed bone healing and nonunion. Bone regeneration is based on the interaction between osteogenesis and angiogenesis. Angiogenesis serves a unique role in the repair and remodeling of bone defects. Monocyte chemoattractant protein-1, also known as CC motif ligand 2 (CCL2), is a member of the CC motif chemokine family and was the first human chemokine to be revealed to be an effective chemokine of monocytes. However, its underlying mechanism in angiogenesis of bone defect repair remains to be elucidated. Therefore, the present study investigated the detailed mechanism by which CCL2 promoted angiogenesis in bone defects based on cell and animal model experiments. In the present study, CCL2 promoted proliferation, migration and tube formation in human umbilical vein endothelial cells (HUVECs) in a concentration-dependent manner. Western blot analysis revealed that treatment of HUVECs with CCL2 upregulated the protein expression levels of rho-associated coiled-coil-containing protein kinase (Rock)1, Rock2, N-cadherin, c-Myc and VEGFR2. Furthermore, CCL2 promoted the expression of MAPK/ERK1/2/MMP9, PI3K/AKT and Wnt/ß-catenin signaling pathway-related proteins, which also demonstrated that CCL2 promoted these functions in HUVECs. Immunohistochemical staining of Sprague Dawley rat femurs following bone defects revealed that VEGF expression was positive in the newly formed bone area in each group, while the expression area of VEGF in the CCL2 addition group was markedly increased. Therefore, CCL2 is a potential therapeutic approach for bone defect repair and reconstruction through the mechanism of angiogenesis-osteogenesis coupling.

Baicalin induces apoptosis and autophagy in human osteosarcoma cells by increasing ROS to inhibit PI3K/Akt/mTOR, ERK1/2 and ß-catenin signaling pathways.

Baicalin, a flavonoid derivative, exerts antitumor activity in a variety of neoplasms. However, whether baicalin exerts antitumor effects on osteosarcoma cells remains to be elucidated. In this study, treatment with baicalin reduced the proliferation and invasive potential of osteosarcoma cells and reduced the mitochondrial membrane potential, which eventually caused mitochondrial apoptosis. In addition, baicalin increased intercellular Ca2+ and ROS concentrations. Baicalin-induced apoptosis was confirmed by enhanced Bax, cleaved caspase-3, and cleaved PARP levels and decreased Bcl-2 levels. The increase in LC3-II and p62 suggested that baicalin induced autophagosome formation but ultimately inhibited downstream autophagy. Moreover, apoptosis induced by baicalin was attenuated by the addition of 3-MA. Furthermore, we found that baicalin inhibited the PI3K/Akt/mTOR, ERK1/2 and ß-catenin signaling pathways. Chelation of free Ca2+ by BAPTA-AM also inhibited both apoptosis induction and ROS concentration changes. Finally, NAC pretreatment reversed baicalin treatment outcomes, including the increase in Ca2+ concentration, induction of apoptosis and autophagy, and inhibition of the pathways. Molecular docking results indicated that baicalin might interact with the structural domain of PI3Kγ. Thus, baicalin may be considered a potential candidate for osteosarcoma treatment.

α-Carotene: a valuable carotenoid in biological and medical research.

α-Carotene, one of the C40 carotenes, is a natural lipid-soluble terpene. The chemical structure of α-carotene is based on the unsaturated polyene chain skeleton, with an ε-ring and a ß-ring on each side of the skeleton. α-Carotene is widely found in dietary fruits and vegetables, and the concentration depends on the plant species. In addition, processing methods and storage conditions used in the food and medical industries can alter the concentration of α-carotene in raw materials. This review of α-carotene summarizes the major studies on chemical structure, source, extraction, detection, biosynthesis, processing effect, bioactivity, medicine, and biotechnology. Whether α-carotene supplementation or a diet rich in fruits and vegetables has a positive effect on the prevention of cancer, cardiovascular disease, and other diseases is the focus of this study. © 2022 Society of Chemical Industry.

Anxiety of Nurses to support Wuhan in fighting against COVID-19 Epidemic and its Correlation With Work Stress and Self-efficacy.

AIMS AND OBJECTIVES: We aimed to investigate the anxiety of nurses who are supporting Wuhan in fighting against coronavirus disease 2019 (COVID-19) infection and explore relevant influencing factors. BACKGROUND: The COVID-19 outbreak poses a major threat to public health worldwide. Nurses play an important role in this epidemic. However, available data on the mental health among these nurses are limited. DESIGN: A descriptive, cross-sectional survey was performed. METHODS: An online questionnaire was completed by 200 nurses who went to Wuhan to help to fight against COVID-19 from another province. Data collection tools include the Chinese version of the Stress Overload Scale (SOS), the Self-Rating Anxiety Scale (SAS) and General Self-Efficacy Scale (GSES). Descriptive, single-factor correlation and multiple regression analyses were used in exploring related influencing factors. Reporting followed the STROBE guidelines. RESULTS: The scores of SAS, SOS and GSES range from 20 to 80, 22 to 110 and 10 to 40, respectively, and the SAS (31.79 ± 7.32) and SOS (40.19 ± 12.92) and GSES scores (24.83 ± 6.60) were obtained. Anxiety was positively correlated with stress (r = .679, p < .001) but negatively correlated with self-efficacy (r = -.326, p < .001). Multiple regression analysis showed that professional qualification, sleep, stress and self-efficacy were the main factors affecting nurse anxiety (p = .006, <.001, <.001, .039, respectively). CONCLUSIONS: Nurses who are supporting Wuhan in fighting against COVID-19 were under a low level of anxiety. RELEVANCE TO CLINICAL PRACTICE: The current study suggests work stress reduction might be a key factor in reducing anxiety and maintaining mental health to support nurses who are fighting against COVID-19 infection.

Herbal therapy treatment in thromboangiitis obliterans: a retrospective clinical study.

BACKGROUND: Critical limb ischemia in patients with thromboangiitis obliterans (TAO, Buerger's disease) is associated with refractory rest pain, gangrene, and increased rates of amputation. Tuoju lotion was prepared by the Pharmacy Department of Dongfang Hospital. The focus of the study is to elicit the efficacy of the addition of Herbal therapy treatment to conventional treatment in TAO patients with severe extremity pain and to assess any statistically significant benefits in patient's pain control at rest. We fund that the addition of herbal therapy treatment can augment conventional treatments in TAO patients by improving or eliminating intermittent claudication symptoms, prolonging claudication distance, and reducing total blood viscosity. At the same time, Tuoju lotion can improve microcirculation status in the short term. The purpose of this study was to investigate the effect of topical Herbal therapy treatment on patient outcomes in patients with TAO. METHODS: Seventy patients with TAO treated between January 2009 and July 2019 were included in a retrospective analysis of a single university hospital vascular center. Forty patients received topical herbal treatment in addition to conventional therapy and were compared to a control group who received standard treatment alone (n=30). RESULTS: Patients in both, the experimental and control group, were matched according to age and gender. There was no significant difference in course of disease and past medical history between the two groups. The mean ankle brachial index (ABI), toe pressure, and blood viscosity were also similar in both groups. Rest pain score (baseline VAS 4.76±2.87, post-treatment 3.32±1.29) and walking distance (baseline 169.7±23.6 m, post-treatment 284.5±32.3 m) significantly improved in the herbal treatment group. ABI values improved and total blood viscosity decreased in both groups with no significant difference between the herbal and conservative treatment arms. However, the arterial blood pressure ratio in the lower extremity stage showed no difference between the superficial femoral artery and the popliteal artery. CONCLUSIONS: The addition of Herbal therapy treatment to conventional treatment in TAO patients with severe extremity pain was associated with a reduction of rest pain and intermittent claudication.

Experimental Study of Phlebitis Ointment Administration in Acute Superficial Thrombophlebitis.

Acute superficial thrombophlebitis is a venous system disease. Animal models with mannitol induced phlebitis were treated with an orally administered "phlebitis ointment." 24 rabbits were randomly divided into 4 groups. The therapy group was treated with "phlebitis ointment" and a control group received "Mai Luo Shu Tong granules." Levels of blood TNF-α, IL-6, CRP, and IL-1ß were measured. The tissue expression levels of NF-КBp65 and PKC genes were evaluated. The therapy group showed a better improvement of the clinical status and similar vascular morphology than the control group. A blank group showed no vascular changes through pathological investigation. In contrast, significant vascular changes were seen in the model group. The control group showed slight vascular modifications. Small thrombi could be found in the lumen despite the intact tunica intima. Both control and therapy group showed less inflammatory cells infiltration than the model group and upregulation of NF-КBp65 and PKC genes. The phlebitis ointment reduced the levels of necrosis factor-α, interleukin-6, C-reactive protein, and interleukin-1ß. The expressions of NF-КBp65 and PKC genes, which are the primary mechanisms underlying the development of thrombophlebitis, were improved significantly in tissues of both therapy group and control group.

Oligothiophene-based colorimetric and ratiometric fluorescence dual-channel cyanide chemosensor: Sensing ability, TD-DFT calculations and its application as an efficient solid state sensor.

An oligothiophene-based colorimetric and ratiometric fluorescence dual-channel cyanide chemosensor 3 T-2CN was reported. Sensor 3 T-2CN showed both naked-eye recognition and ratiometric fluorescence response for CN- with an excellent selectivity and high sensitivity. The sensing mechanism based on the nucleophilic attack of CN- on the vinyl CC bond has been successfully confirmed by the optical measurements, 1H NMR titration, FT-IR spectra as well as the DFT/TD-DFT calculations. Moreover, the detection limit was calculated to be 0.19µM, which is much lower than the maximum permission concentration in drinking water (1.9µM). Importantly, test strips (filter paper and TLC plates) containing 3 T-2CN were fabricated, which could act as a practical and efficient solid state optical sensor for CN- in field measurements.

[Effect of effective components of Huanglian Jiedu decocting on NF-kappaB in cultured rat cerebral microvascular endothelial cells injury induced by hypoxia and reoxygenation].

OBJECTIVE: To investigate the protective mechanism of geniposide, baicalin and berberine on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells. METHOD: To establish a model of hypoxia four hours and reoxygenation twelve hours injury in rat cerebral microvascular endothelial cells in vitro. The injured cells were treated with geniposide (0. 128, 0.064, 0.032 micromol mL(-1), baicalin (0.028, 0.014, 0.007 micromol mL(- 1)) and berberine (0.024, 0.012, 0.006 micromol mL(-1)). The expression of p65 subunit of NF-kappaB was detected by immunocytochemical assay and techniques of image quantitative analysis. The protein expression of NF-kappaB was calculated with the mean optical density and mean area. The nuclear translocation of NF-kappaB was calculated with the percentage of positive cells and ratios of light transmittance of cytoplasm and cell nucleus. RESULT: Compared with the normal group, both the protein expression and the nuclear translocation of NF-kappaB of model group were significant increased (P <0.01). Compared with the model group, the mean optical density of all treated groups was decreased ,but these was no significant difference between them. As compared with model group, the mean area of all treated groups was significant decreased (P < 0.01). The percentage of nuclear translocation of all treated groups is not only lower than that of the model group but higher than that of the normal group (P <0.01). Compared with the model group, the ratios of light transmittance of cytoplasm and cell nucleus of all treated groups was significantly elevated (P <0.01). CONCLUSION: The results suggesed that geniposide, baicalin and berberine could protect hypoxia/reoxygenation injuried rat cerebral microvascular endothelial cells injury. One of the mechanism may lie in inhibiting both the protein expression and the nuclear translocation of NF-kappaB.

[Effect of effective components of huanglian jiedu decoction on hypoxia, reoxygenation injury and the expression of VCAM in cultured rat cerebral microvascular endothelial cells].

OBJECTIVE: To investigate the protective mechanism of geniposide, baicalin and berberine on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells. METHOD: A model of four hours hypoxia and twelve hours reoxygenation injury in rat cerebral microvascular endothelial cells in vitro was established. The injured cells were treated with geniposide (0.128, 0.064, 0.032 mmol x L(-1)), baicalin (0.028, 0.014, 0.007 mmol L(-1)) and berberine (0.024, 0.012, 0.006 mmol L(-1)), respectively. The immunocytochemical method and techniques of image quantitative analysis were used to detect the mean optical density and mean area in order to match the protein expression of VCAM-1. The method of RT-PCR was adopted to observe and match the mRNA expression of VCAM-1. RESULT: As compared with the normal group, the mean optical density, the mean area and the mRNA expression of VCAM-1 of model group were significant increased (P < 0.01, P < 0.01, P < 0.01). As compared with the model group, both the mean optical density and the mean area of all treated groups were decreased, and there was significant difference between them (P < 0.01, P < 0.01). As compared with normal group, the mean optical density of baicalin (0.007 mmol x L(-1)) and berberine (0.012, 0.006 mmol x L(-1)) were significant decreased (P < 0.05, P < 0.01, P < 0.01), but there was no significant difference between the other groups and the normal group. As compared with normal group, the mean area of baicalin (0.0014 mmol x L(-1)) was significant decreased (P < 0.05), but there was significant difference between the other groups and the normal group. The mRNA expression of all treated groups was not only lower than that of the model group but also higher than that of the normal group (P < 0.05, P < 0.05). CONCLUSION: The results suggest that geniposide, baicalin and berberine, which are effective compositions of huanglian jiedu decoting, can protect hypoxia-reoxygenation injuried rat cerebral microvascular endothelial cells. One of the protected mechanisms is that they can inhibit the expression of VCAM-1.

[Protective action of effective components of Huanglian Jiedu decoction on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells].

OBJECTIVE: To investigate the protective effect of geniposide, baicalin and berberine for the rat cerebral microvascular endothelial cell. METHOD: The model of hypoxia and reoxygenation injury in rat cerebral microvascular endothelial cells in vitro was established. Both normal and model cells were treated with geniposide (1.024, 0.512, 0.256, 0.128, 0.064, 0.032, 0.016, 0.008 micromol x mL(-1)), baicalin (0.224, 0.112, 0.056, 0.028, 0.014, 0.007, 0.003 micromol x mL(-1)) and berberine (0.192, 0.096, 0.048, 0.024, 0.012, 0.006, 0.003 micromol x mL(-1)). Cell activity was measured by methyl thiazolyl tetrazolium (MTT) test. RESULT: After hypoxia/hypoglycemia cultures for 4 hour and reoxygenation for 12 hour, geniposide (0.128, 0.064, 0.032 micromol x mL(-1)), baicalin (0.028, 0.014, 0.007 micromol x mL(-1)) and berberine (0.024, 0.012, 0.006 micromol x microL(-1) could protect the injuried cerebral microvascular endothelial cells. CONCLUSION: Appropriate concentration of geniposide, baicalin and berberine, which are effective components of Huanglian Jiedu decoction, could protect the injuried cerebral microvascular endothelial cells.

[Protect effects of Qingkailing injection on mitochondrion membrane potential during injury induced by hypoxia-hypoglycemia and reoxygenation in cultured rat hippocampal neurons].

OBJECTIVE: To investigate the protect effects of Qingkailing injection on mitochondrion membrane potential (MMP) during injury induced by hypoxia-hypoglycemia and reoxygenation in cultured rat hippocampal neurons. METHOD: Mitochondrion activity was measured by methyl thiazolyl tetrazolium (MTT) test. MMP and apoptosis were measured by flow cytometry. intracellular free calcium concentration ([Ca2+]i) was measured with confocal laser scanning microscopy. RESULT: Hypoxia-hypoglycemia cultures for 5 hour and reoxygenation for 3 hour induced intracellular[Ca2+]i and apoptosis rate significantly increased. The effects were increased with the extending time of reoxygenation. MMP and mitochondrion activity declined significantly after 3 hour reoxygenation. The effects were declined with the extending time of reoxygenation. Qingkailing injection might have significantly decrease intracellular [Ca2+]i and apoptosis rate, increase MMP and mitochondrion activity. CONCLUSION: Qingkailing Injection might have significantly inhibit the decline in MMP induced by hypoxia-hypoglycemia and reoxygenation, and had effects of stable it and anti-neuronal apoptosis. The effects might be related to inhibit overload of intracellular free calcium.