Mechanism of Smilax china L. in the treatment of intrauterine adhesions based on network pharmacology, molecular docking and experimental validation.

BACKGROUND: Smilax china L. (SCL) is a traditional herbal medicine for the potential treatment of intrauterine adhesion (IUA). However, the mechanisms of action have not yet been determined. In this study, we explored the effects and mechanisms of SCL in IUA by network pharmacology, molecular docking and molecular biology experiments. METHODS: Active ingredients and targets of SCL were acquired from TCMSP and SwissTargetPrediction. IUA-related targets were collected from the GeneCards, DisGeNET, OMIM and TTD databases. A protein‒protein interaction (PPI) network was constructed by Cytoscape 3.9.1 and analysed with CytoHubba and CytoNCA to identify the core targets. The DAVID tool was used for GO and KEGG enrichment analyses. Furthermore, molecular docking was employed to assess the interaction between the compounds and key targets. Finally, the mechanisms and targets of SCL in IUA were verified by cellular experiments and western blot. RESULTS: A total of 196 targets of SCL were identified, among which 93 were related to IUA. Topological and KEGG analyses results identified 15 core targets that were involved in multiple pathways, such as inflammation, apoptosis, and PI3K/AKT signalling pathways. Molecular docking results showed that the active compounds had good binding to the core targets. In vitro experiments showed that astilbin (AST), a major component of SCL, significantly reduced TGF-ß-induced overexpression of fibronectin (FN), activation of the PI3K/AKT signalling pathway and the expression of downstream factors (NF-κB and BCL2) in human endometrial stromal cells, suggesting that AST ameliorates IUA by mediating the PI3K/AKT/NF-κB and BCL2 proteins. CONCLUSIONS: AST, a major component of SCL, may be a potential therapeutic agent for IUA. Moreover, its mechanism is strongly associated with regulation of the PI3K/AKT signalling pathway and the downstream NF-κB and BCL2 proteins. This study will provide new strategies that utilize AST for the treatment of IUA.

Waterlogging in soil restricts the growth of Gleditsia sinensis seedlings and inhibits the accumulation of lignans and phenolic acids in thorns.

Gleditsia sinensis, commonly known as Chinese Zaojiao, has important economic value and medicinal compounds in its fruits and thorns, making it widely cultivated artificially in China. However, the available literature on the impact of waterlogging on the growth of G. sinensis seedlings and the accumulation of metabolite compounds in its thorns is limited. To address this knowledge gap, G. sinensis seedlings were planted in soil supplemented with pindstrup substrate, which enhances the water-holding capacity of the soil. The analyses of morphological traits and nutrient elements in one-year-old G. sinensis seedlings grown naturally under ambient conditions and metabolite accumulation in its thorns were conducted. The results showed that the waterlogged soil significantly diminished the height, fresh weight, and dry weight of seedling roots and stems (P < 0.05). Furthermore, waterlogging hindered the uptake of iron (Fe) and manganese (Mn), as well as the transport of potassium (K). The identified metabolites within the thorns were categorized into 16 distinct groups. Relative to the control soil, fatty acids and derivatives were the most down-regulated metabolites in the waterlogged soil, accounting for 40.58% of the total metabolites, followed by lignans (38.71%), phenolic acids (34.48%), saccharides and alcohols (34.15%), steroids (16.67%), alkaloids (12.24%), flavonoids (9.28%), and glycerophospholipids (7.41%). Conversely, nucleotides and derivatives experienced the greatest up-regulation in the waterlogged soil, accounting for 50.00% of the total metabolites. In conclusion, waterlogging negatively impacted the growth of G. sinensis seedlings and inhibited the accumulation of metabolites. Hence, when considering the accumulation of secondary metabolites such as lignans and phenolic acids, appropriate management of soil moisture levels should be taken into account.

Improving Blood Pressure Control and Pregnancy Outcomes: The Impact of Evidence-Based Care and Aerobic Exercise in Patients with Hypertensive Disorders During Pregnancy.

Objective: To explore the effect of evidence-based care plus aerobic exercise on blood pressure control and pregnancy outcome in patients with hypertensive disorders during pregnancy. Methods: A total of 100 patients diagnosed with hypertensive disorder in pregnancy treated in our hospital between February 2020 and November 2021 were recruited, analyzed and assigned at a ratio of 1:1 to receive routine nursing (control group) or evidence-based care plus aerobic exercise (experimental group) via random number table method. Outcome measures included blood pressure, negative emotions, sleep duration, and pregnancy outcome. Results: The blood pressure of both groups decreased after nursing, and the diastolic and systolic blood pressure of patients in the experimental group (79.84±5.18 mmHg, 111.62±7.96 mmHg) were lower than those in the control group (88.65±5.69 mmHg, 132.15±8.14 mmHg) (P < .05). After the completion of the nursing period, assessments using the Self-Rating Anxiety Scale and Hamilton Depression Scale were conducted. The results revealed significantly lower scores in the experimental group, which received evidence-based care along with aerobic exercise, compared to the control patients who received routine care. The sleep duration was prolonged in both groups after nursing, and patients in the experimental group got longer sleep duration than those in the control group (P < .05). The experimental group showed a significantly lower incidence of adverse pregnancy outcomes than the control group (P < .05). Limitations: While our study demonstrates the positive impact of evidence-based care combined with moderate aerobic exercise on patients with hypertensive disorders during pregnancy, it is essential to acknowledge some notable limitations. First, the sample size was relatively small, which may limit the generalizability of our findings to a larger population. Furthermore, our study primarily focused on short-term outcomes, and future research could explore the sustained benefits of this approach. Finally, individual variations in exercise tolerance and compliance may also affect the effectiveness of the intervention. Despite these limitations, our findings hold promise and provide a foundation for further research in this area. Conclusion: Evidence-based care combined with moderate aerobic exercise has proven to be an effective approach in enhancing the overall management of patients with hypertensive disorders during pregnancy. This combined intervention not only effectively regulates blood pressure levels but also mitigates adverse emotional states, enhances sleep quality, and ultimately leads to improved pregnancy outcomes. These findings hold significant promise for clinical application. Healthcare providers may consider implementing this approach to improve the well-being of pregnant individuals with hypertensive disorders, potentially reducing the risk of complications and enhancing the overall quality of care. Pregnant individuals, on the other hand, can benefit from a more comprehensive and holistic approach to their care, which may result in better health and pregnancy outcomes. Future research in this area could explore the long-term sustainability and cost-effectiveness of this intervention, as well as its potential applicability to diverse patient populations and healthcare settings.

Efficacy of Quality Care on Maternal and Infant Outcomes in Patients with Hypertensive Disorders Complicating Pregnancy Complicated with Cerebral Hemorrhage.

Objective: This study aimed to evaluate the impact of quality care on maternal and infant outcomes in patients with hypertensive disorders complicating pregnancy (HDCP) complicated by cerebral hemorrhage. Methods: From February 2020 to September 2021, 68 women with HDCP complicated by cerebral hemorrhage hospitalized at our hospital were included and divided into a routine group (standard care) and a quality group (quality care). Outcome measures included National Institutes of Health Stroke Scale (NIHSS) scores, blood pressure, self-rating anxiety scale (SAS) scores, self-rating depression scale (SDS) scores, and maternal and infant outcomes. Results: Patients in the quality group (3.22±1.89) had significantly lower NIHSS scores aftercare than those in the routine group (6.15±3.24) (P < .05). Quality care resulted in lower diastolic blood pressure (Quality group:81.23±6.15; Routine: 90.58±7.98), systolic blood pressure (Quality group:125.49±13.37; Routine: 139.74±16.67), SAS scores (Quality group: 48.42±2.65; Routine: 58.15±2.43), and SDS scores versus routine care (Quality group:48.42±2.65; Routine: 58.15±2.43)(P < .05). The quality group showed a lower incidence of adverse maternal and infant pregnancy outcomes than the routine group (P < .05). Conclusion: The findings underscore the positive impact of quality care in reducing adverse maternal and newborn pregnancy outcomes. This reduction is particularly significant for clinical practice, as it is achieved through the amelioration of various factors, such as neurological impairments, blood pressure regulation, and the alleviation of negative emotions, including anxiety and depression, in patients with HDCP complicated by cerebral hemorrhage. The practical implications of these findings for healthcare providers and patients are substantial. They highlight the potential to improve patient outcomes, enhance the overall quality of care, and reduce the burden on healthcare systems. By addressing these factors, healthcare providers can enhance the well-being of both mothers and newborns, leading to improved clinical outcomes and increased patient satisfaction.

Wogonin attenuates inflammation and oxidative stress in lipopolysaccharide-induced mastitis by inhibiting Akt/NF-κB pathway and activating the Nrf2/HO-1 signaling.

Mastitis is a disease involved in inflammation of breast which affects human and animals. Wogonin is one bioactive compound from many Chinese herbal medicines, which have multiple properties, including anti-inflammatory activity. However, the roles of wogonin in mastitis progression are largely undefined. Mastitis models were established using LPS-treated mice and mammary epithelial cells (MECs). Infiltration of inflammatory cells was analyzed by hematoxylin-eosin staining and myeloperoxidase (MPO) activity. Inflammatory cytokine (TNF-α and IL-1ß) levels were detected via ELISA. The phosphorylation and total of Akt and NF-κB levels and content of Nrf2 and HO-1 were measured via western blot. Cell viability was examined by CCK-8 assay. Oxidative stress was assessed by ROS generation and levels of MDA, GSH, and SOD. Wogonin attenuated LPS-induced infiltration of inflammatory cells, increase of MPO activity and levels of TNF-α and IL-1ß, and activation of the Akt/NF-κB pathway in murine mammary gland tissues, and promoted activation of Nrf2/HO-1 signaling. Wogonin did not affect MEC viability, but mitigated LPS-induced inflammation in MECs by reducing TNF-α and IL-1ß levels. Wogonin relieved LPS-induced oxidative stress in MECs through decreasing ROS generation and MDA level and increasing GSH and SOD levels. Wogonin repressed LPS-induced activation of the Akt/NF-κB pathway in MECs and increased Nrf2/HO-1 signaling activation. Activated Akt/NF-κB signaling or Nrf2/HO-1 signaling inactivation reversed the suppressive effects of wogonin on LPS-induced inflammation and oxidative stress in MECs. Wogonin mitigates LPS-induced inflammation and oxidative stress of MECs via suppressing activation of the Akt/NF-κB signaling and activating Nrf2/HO-1 pathway, indicating the therapeutic potential of wogonin in mastitis.

Rapid Isolation and Hypoglycemic Activity of Secondary Metabolites of Eurotium cristatum by High-Speed Countercurrent Chromatography.

In this study, secondary metabolites of Eurotium cristatum were isolated and purified by high-speed counter-current chromatography (HSCCC), and their hypoglycemic activities were studied. The general-useful estimate of solvent systems (GUESS) for counter-current chromatography was employed to select the appropriate solvent systems of n-hexane-ethyl acetate-methanol-water (HEMW, 4:6:5:5, v/v/v/v) for HSCCC practice, and three compounds were separated from the crude ethyl acetate extract of E. cristatum in one single step; 6.1 mg of Compounds 1, 5.6 mg of Compound 2 and 3.8 mg of Compound 3 were obtained from 100 mg of crude extract with a stationary phase retention of 75%. The compounds were then identified as emodin methyl ether, chrysophanol and emodin, respectively. The activity of the target compounds in the secondary metabolites of E. cristatum was verified by testing their inhibition on α-glucosidase activity and molecular docking simulation. The results showed that emodin, chrysophanol and emodin methyl ether had significant inhibitory effects on the α-glucosidase activity. This work confirmed the effectiveness of HSCCC in the separation of compounds in complex extracts and provided reference for further research and application of E. cristatum.

Acupuncture Needle-Based Transistor Neuroprobe for In Vivo Monitoring of Neurotransmitter.

Chemical communication via neurotransmitters is central to brain functions. Nevertheless, in vivo real-time monitoring of neurotransmitters released in the brain, especially the electrochemically inactive molecules, remains a great challenge. In this work, a novel needle field-effect transistor (FET) microsensor based on an acupuncture needle is proposed, which is demonstrated to be capable of real-time monitoring dopamine molecules as well as neuropeptide Y in vivo. The FET microstructure is fabricated by successively wrapping an insulating layer and a gold layer on the top of the needle, where the needle and the Au served as the source and drain, respectively. After assembling reduced graphene oxide (RGO) between the source and drain electrodes, the specific aptamer is immobilized on the RGO, making this needle-FET biosensor highly selective and sensitive to real-time monitor neurotransmitters released from rat brain, even in a Parkinson's diseases model. Furthermore, the needle-FET biosensor is applied to detect a variety of targets including hormones, proteins, and nucleic acid. By constructing a FET sensing interface on an acupuncture needle and implanting the sensor in a rat's brain for in vivo detection, this work provides a new sight in the FET domain and further expands the species of real-time in vivo detection.

Cationic amino acid transporter-1 (CAT-1) promotes fibroblast-like synoviocyte proliferation and cytokine secretion by taking up L-arginine in rheumatoid arthritis.

BACKGROUND: Abnormal proliferation of fibroblast-like synoviocytes (FLSs) in the synovial lining layer is the primary cause of synovial hyperplasia and joint destruction in rheumatoid arthritis (RA). Currently, the relationship between metabolic abnormalities and FLS proliferation is a new focus of investigation. However, little is known regarding the relationship between amino acid metabolism and RA. METHODS: The concentrations of amino acids and cytokines in the synovial fluid of RA (n = 9) and osteoarthritis (OA, n = 9) were detected by LC-MS/MS and CBA assay, respectively. The mRNA and protein expression of cationic amino acid transporter-1 (CAT-1) were determined in FLSs isolated from RA and OA patients by real-time PCR and western blotting. MTT assay, cell cycle, apoptosis, invasion, and cytokine secretion were determined in FLSs knocked down of CAT-1 using siRNA or treated with D-arginine under normoxic and hypoxic culture conditions. A mouse collagen-induced arthritis (CIA) model was applied to test the therapeutic potential of blocking the uptake of L-arginine in vivo. RESULTS: L-rginine was upregulated in the synovial fluid of RA patients and was positively correlated with the elevation of the cytokines IL-1ß, IL-6, and IL-8. Further examination demonstrated that CAT-1 was the primary transporter for L-arginine and was overexpressed on RA FLSs compared to OA FLSs. Moreover, knockdown of CAT-1 using siRNA or inhibition of L-arginine uptake using D-arginine significantly suppressed L-arginine metabolism, cell proliferation, migration, and cytokine secretion in RA FLSs under normoxic and hypoxic culture conditions in vitro but increased cell apoptosis in a dose-dependent manner. Meanwhile, in vivo assays revealed that an L-arginine-free diet or blocking the uptake of L-arginine using D-arginine suppressed arthritis progression in CIA mice. CONCLUSION: CAT-1 is upregulated and promotes FLS proliferation by taking up L-arginine, thereby promoting RA progression.

[Effect of manual acupuncture at "Sanyinjiao" (SP6) on spermatogenic cells of mice with oligoasthenospermia induced by microwave radiation].

OBJECTIVE: To observe the effect of acupuncture at "Sanyinjiao" (SP6) on sperm motility, testicular B cell lymphoma/leukelia-2 (Bcl-2), Bcl-2 associated X protein (Bax), and Caspase-3 in mice with oligoasthenospermia induced by microwave radiation, so as to explore its underlying mechanisms in improving oligoasthenospermia. METHODS: Male BALB/C mice were randomly divided into control, model and acupuncture groups(n=6 in each group). The oligoasthenospermia model was established by continuous microwave irradiation with frequency of 2 450 MHz and power density of 40 mW/cm2, 1 h daily for 18 days. At the same time, manual acupuncture was applied to the acupuncture group on bilateral "Sanyinjiao" (SP6) for 30 s, once daily for 18 days. Sperm motility including the percentages of progressive motility (PR), non-progressive motility (NP), and PR + NP sperms was detected by computer-assisted sperm analysis, H.E. staining was used to observe the testicular morphology and Johnson score was calculated, the expression levels of Bcl-2, Bax and Caspase-3 in testis were detected by immunofluorescence. RESULTS: Compared with the control group, the percentages of PR sperms, NP sperms, PR+NP sperms, Johnson score, and expression level of Bcl-2 were significantly decreased (P<0.05), while the expression levels of Bax and Caspase-3 were increased (P<0.05) in the model group. Compared with the model group, the percentages of PR sperms, PR+NP sperms, Johnson score, and expression level of Bcl-2 were significantly increased (P<0.05), while the expression levels of Bax and Caspase-3 were decreased (P<0.05) in the acupuncture group. Outcomes of H.E. staining showed that the seminiferous tubules became thinner, spermatogenic cells and sperm decreased or even disappeared, and the supporting cells were partially missing in the model group, which was relatively milder in the acupuncture group. CONCLUSION: Manual acupuncture at SP6 can improve sperm motility in oligoasthenospermia mice induced by microwave radiation, which may be related to its effects in down-regulating the expressions of Bax and Caspase-3, increasing expression of Bcl-2 in the testis.

Enzymatic Biotransformation of Gypenoside XLIX into Gylongiposide I and Their Antiviral Roles against Enterovirus 71 In Vitro.

Biotransformation of specific saponins in the valuable medical plants to increase their bioavailability and pharmaceutical activities has attracted more and more attention. A gene encoding a thermophilic glycoside hydrolase from Fervidobaterium pennivorans DSM9078 was cloned and expressed in Escherichia coli. The purified recombinant enzyme, exhibiting endoglucanase cellulase activity, was used to transform gypenoside XLIX into gylongiposide I via highly selective and efficient hydrolysis of the glucose moiety linked to the C21 position in gypenoside XLIX. Under the optimal reaction conditions for large scale production of gylongiposide I, 35 g gypenoside XLIX was transformed by using 20 g crude enzyme at pH 6.0 and 80 °C for 4 h with a molar yield of 100%. Finally, 11.51 g of gylongiposide I was purified using a silica gel column with 91.84% chromatographic purity. Furthermore, inhibitory activities of gypenoside XLIX and gylongiposide I against Enterovirus 71 (EV71) were investigated. Importantly, the EC50 of gypenoside XLIX and gylongiposide I calculated from viral titers in supernatants was 3.53 µM and 1.53 µM, respectively. Moreover, the transformed product gylongiposide I has better anti-EV71 activity than the glycosylated precursor. In conclusion, this enzymatic method would be useful in the large-scale production of gylongiposide I, which would be a novel potent anti-EV71 candidate.

Ultrasound Multiparametric Assessment of the Impact of Hypertensive Disorders of Pregnancy on Fetal Cardiac Function and Growth and Development.

Objective: To evaluate the ultrasound multiparametric assessment of the impact of hypertensive disorders of pregnancy (HDP) on fetal cardiac function and growth and development. Methods: In this prospective study, 98 cases of HDP treated in our institution were recruited into a study group, and 100 pregnant women with healthy singleton pregnancies were included in a control group. All eligible patients were also assigned to either study group A (HDP fetuses with growth restriction) or study group B (HDP fetuses with normal growth). Fetal echocardiography was performed on all eligible participants to obtain hemodynamic and cardiac function parameters for the evaluation of fetal growth and development, and the impact of HDP on fetal heart function and growth and development was analyzed. Results: HDP fetuses were associated with smaller head circumference, biparietal diameter, femoral length, and abdominal circumference versus healthy fetuses. The study group had a higher resistance index (RI) and pulsatility index (PI) of umbilical artery (UA), ductus venous (DV), pulmonary vein (PV), and lower RI and PI of aortic isthmus (AoI) than the control group. The study group showed higher left and right ventricular isovolumic contraction time (IVCT), isovolumic relaxation time (IVRT), and myocardial performance index (MPI) values and lower mitral and tricuspid E wave and E/A values than the control group. The systolic blood pressure was positively correlated with PI, RI of UA, DV, and PV, and left and right ventricular IVCT, IVRT, and MPI and negatively correlated with PI and RI of AoI and mitral and tricuspid E wave and E/A values of HDP fetuses. The peak systolic/diastolic flow rate (S/D), PI, and RI of umbilical blood flow in study group A were higher than those in study group B. Umbilical blood flow S/D showed the highest AUC and specificity for predicting fetal growth restriction, and PI had the highest sensitivity for predicting fetal growth restriction. Conclusion: HDP compromises fetal cardiac function and growth, and ultrasound multiparametric assessment provides accurate detection of fetal cardiac function and hemodynamics changes. The patient's condition can be monitored through the assessment of ultrasound parameters of fetal growth and development.

Ultrasound of Fetal Cardiac Function Changes in Pregnancy-Induced Hypertension Syndrome.

Pregnancy-induced hypertension syndrome (PIH) is a common pregnancy syndrome that could cause varying degrees of maternal and fetal organic damage and even endanger their lives. This study aimed to investigate ultrasound of fetal cardiac function changes in PIH. Totally 40 cases of gestational hypertension admitted to Cangzhou Central Hospital between October 2018 and September 2019 were enrolled in the hypertension group, and 40 women with healthy pregnancies during the same period were assigned to the normal group. Ultrasound results showed that PIH was associated with a significantly higher fetal cardiac septal thickness, fetal left ventricular end-diastolic area and end-systolic area, fetal right ventricular end-diastolic area and end-systolic area, fetal left ventricular systolic fraction 1 (VSF1), fetal left ventricular systolic fraction 2 (VSF2), fetal right VSF1, and fetal right VSF2 versus healthy pregnancy. The PIH fetuses had significantly lower neonatal weights versus healthy fetuses. Newborns of hypertensive pregnancies have larger hearts, faster heart rates, increased cardiac contractility, and lower weights versus newborns of healthy pregnancies.

Effect of advanced nursing care on psychological condition in patients with chronic heart failure: A protocol of systematic review and meta-analysis.

BACKGROUND: This study will appraise the effect and safety of advanced nursing care (ANC) on psychological condition (PC) in patients with chronic heart failure (CHF). METHODS: The following databases will be sought from the beginning up to the February 29, 2020: MEDLINE, EMBASE, Cochrane Library, Web of Science, Scopus, the Cumulative Index to Nursing and Allied Health Literature, the Allied and Complementary Medicine Database, the Chinese Scientific Journal Database, and China National Knowledge Infrastructure. There are not language and publication status limitations related to any electronic databases. In addition, we will also identify conference proceedings, reference lists of included studies, and websites of clinical trials registry. Two reviewers will separately carry out study selection, data extraction, and study quality evaluation. Any inconsistencies will be solved by a third reviewer through discussion. RevMan 5.3 software will be utilized to carry out statistical analysis. RESULTS: This study will comprehensively summarize all potential evidence to systematically address the effects and safety of ANC on PC in patients with CHF. CONCLUSION: The findings of the present study will help to determine whether ANC is effective or not on PC in patients with CHF. STUDY REGISTRATION NUMBER: INPLASY202040077.

Silver-Laden Black Phosphorus Nanosheets for an Efficient In Vivo Antimicrobial Application.

Nanobactericides represent one of the most efficient and promising strategies for eliminating bacterial infection considering the increasing resistance threats of conventional antibiotics. Black phosphorus (BP) is the most exciting postgraphene layered 2D nanomaterial with convincing physiochemical properties, yet the study of BP-based antibiotics is still in its infancy. Here, a compact silver nanoparticle (AgNP)-doped black phosphorus nanosheet (BPN) is constructed to synergistically enhance solar disinfection through the promoted reactive oxygen species (ROS) photogeneration, which is attributed to the improved electron-hole separation and recombination of BPNs as revealed from the systematic experimental studies. An in-depth density functional theory (DFT) calculation confirms that the integrated AgNPs provide a preferred site for facilitating the adsorption and activation of O2 , thus promoting the more efficient and robust ROS generation on BPN-AgNP nanohybrids. Besides the enhanced photoinduced ROS, the anchored AgNPs simultaneously lead to a dramatically increased affinity toward bacteria, which facilitates a synergetic pathogen inactivation. Significantly, the convincing antimicrobial BPN-AgNP contributes to the prominent wound healing and antimicrobial ability in vivo with minimized biological burden. This sophisticated design of new 2D nanohybrids opens a new avenue for further exploiting BP-based nanohybrids in portable bandage and broad-spectrum disinfection applications.

Mechanism of antidiabetic and synergistic effects of ginseng polysaccharide and ginsenoside Rb1 on diabetic rat model.

Ginseng polysaccharides (GP) have been reported to modulate gut microbiota, and ginsenoside Rb1 is known to display significant hypoglycemic activity. However, the synergistic effect of Rb1 and GP when applied to diabetic treatment remains largely unknown. Male rats were divided into ten groups: blank group (B-Group), model group (D-Group), Rb1 group (Rb1-Group), CK group (CK-Group), GP groups and GP + Rb1 groups in dosage of high, middle and low (H-Group, M-Group, L-Group, H-Rb1-Group, M-Rb1-Group, and L-Rb1-Group). CK-Group, GP groups and Rb1 group were fed CK, GP and Rb1 for 30 days, respectively. GP + Rb1 groups were fed GP on the initial 15 days and GP and Rb1 on the final 15 days. The fasting glucose of all groups was measured every five days. The transformation of Rb1 in vitro by rat intestinal microflora, which was collected from the B-Group, D-Group and GP groups on the 15th day, was investigated using HPLC and RRLC-Q-TOF/MS. Analyses the of 16S rRNA gene of the fecal bacterial population and fecal ß-glucosidase activity were conducted among the B-Group, D-Group and H-Group. Compared with those of rats in the D-Group, the fasting glucose levels of rats in the CK-Group and H-Rb1-Group decreased highest. During transformation of Rb1 by diabetic rat intestinal microflora, five transformed products, including ginsenoside Rd, F2, CK, gypenoside XVII (G-XVII), and LXXV (G-LXXV), as well as three transformation pathways, were identified. When a high dose of GP was fed to diabetic rats for 15 days, the formation of intermediates, including G-XVII and G-LXXV was inhibited, and only one pathway (Rb1→Rd→F2→CK) was identified. Moreover, the biotransformation rate of CK increased from 14.0% to 86.7% after 8 h of cultivation. GP reinstated the perturbed holistic gut microbiota and promoted fecal ß-d-glucosidase activity. Ginsenoside Rb1 and GP shows synergistic effects when applied to diabetic treatment and may be developed as a potential antidiabetic drug.

Characterization of steryl glycosides in marine microalgae by gas chromatography-triple quadrupole mass spectrometry (GC-QQQ-MS).

BACKGROUND: Steryl glycosides (SGs) are sterol conjugates found in various plants, especially in those making up human diets. It has been demonstrated that SGs have potential health benefits, and they could be used as food supplements in a variety of food matrixes. Marine microalgae are a potential resource for human food and ingredients. In this study, gas chromatography-triple quadrupole mass spectrometry (GC-QQQ-MS) was used to characterize unknown SGs in eight microalgae belonging to different classes (Isochrysis galbana 3011, Pavlova viridis, Platymonas helgolandica, Conticribra weissflogii, Thalassiosira pseudonana, Nitzschia closterium, Gymnodinium sp., and Karlodinum veneficum). RESULTS: The SGs were first extracted from lyophilized algae with chloroform-methanol, purified by solid-phase extraction and analyzed as trimethylsilyl derivatives. Nine SGs have been identified. In particular, new SGs like occelasteryl glycoside and stellasteryl glycoside were found in Gymnodinium sp., 24-methylene cholesteryl glycoside was detected in P. helgolandica, and 4,24-dimethylcholestan-3-yl glycoside was identified as the main constituent of microalga K. veneficum. The results also showed that the compositions of SGs in different microalgae varied, with a range of 5.234 to 0.036 g kg-1 , and microalga P. viridis contained the most abundant SGs. CONCLUSION: GC-QQQ-MS is a powerful tool to detect SGs with different structures from a variety of microalgae. The compositions of SGs in different microalgae varied greatly. Microalgae are a good source of highly valued SGs. © 2017 Society of Chemical Industry.

Microbial transformation of ginsenoside Rb1, Re and Rg1 and its contribution to the improved anti-inflammatory activity of ginseng.

Microbial transformation of ginsenosides to increase its pharmaceutical effect is gaining increasing attention in recent years. In this study, Cellulosimicrobium sp. TH-20, which was isolated from soil samples on which ginseng grown, exhibited effective ginsenoside-transforming activity. After protopanaxadiol (PPD)-type ginsenoside (Rb1) and protopanaxatriol (PPT)-type ginsenosides (Re and Rg1) were fed to C. sp. TH20, a total of 12 metabolites, including 6 new intermediate metabolites, were identified. Stepwise deglycosylation and dehydrogenation on the feeding precursors have been observed. The final products were confirmed to be rare ginsenosides Rd, GypXVII, Rg2 and PPT after 96 h transformation with 38-96% yields. The four products showed improved anti-inflammatory activities by using lipopolysaccharide (LPS)-induced murine RAW 264.7 macrophages and the xylene-induced acute inflammatory model of mouse ear edema. The results indicated that they could dramatically attenuate the production of TNF-α more effectively than the precursors. Our study would provide an example of a unique and powerful microbial cell factory for efficiently converting both PPD-type and PPT-type ginsenosides to rare natural products, which extends the drug candidates as novel anti-inflammatory remedies.

Wogonin prevents TLR4-NF-κB-medicated neuro-inflammation and improves retinal ganglion cells survival in retina after optic nerve crush.

Chronic neuro-inflammation is involved in the death of retinal ganglion cells (RGCs) in glaucoma. The aim of this study is to determine whether wogonin can suppress inflammatory responses and rescue RGCs death after optic nerve crush (ONC), an ideal animal model of glaucoma. Wogonin was administered intraperitoneally 10 min after establishment of ONC model. In this study, wogonin treatment reduced RGCs loss and inhibited RGCs apoptosis demonstrated by the increased Brn3a labeling RGCs at day 14 and the decreased cleaved caspase-3 expression at day 7 after ONC, respectively. In ONC model, number of GFAP-positive glial cells and iba1-positive microglial cells were increased, combined of the elevated level of pro-inflammatory cytokines released in retina at day 7. However, most of these responses were inhibited after wogonin treatment. The level of TLR4 expression, NF-κB-P65 nucleus location and NF-κB-P65 phosphorylation were increased in retina at day 1 after ONC, which was significantly reduced after wogonin treatment. These results demonstrated that wogonin protected RGCs survival and suppressed neuro-inflammation in retina after ONC by inhibiting TLR4-NF-κB pathways. We conclude that wogonin could be a possible strategy for the treatment of glaucoma.

Tetramethylpyrazine protects CoCl2-induced apoptosis in human umbilical vein endothelial cells by regulating the PHD2/HIF/1α-VEGF pathway.

Tetramethylpyrazine (TMP), one of the active ingredients isolated from a Chinese herbal prescription, possesses protective effects against apoptosis in endothelial cells. However, the underlying mechanism of its protective effects in endothelial cells remains to be elucidated. Using human umbilical vein endothelial cells (HUVECs), the present study assessed the protective effects of TMP on CoCl2-induced apoptosis. Following pre-incubation with CoCl2 (150 µM/ml) for 4 h, the HUVECs were treated with TMP at different concentrations (50, 100 and 200 µM/ml) for 8 h. TMP upregulated the expression of prolyl hydroxylase (PHD)2, reduced the protein and mRNA expression levels of vascular endothelial growth factor (VEGF), and reduced the expression of HIF-1α only at the protein level, not at the mRNA level in HUVECs, in a concentration-dependent manner. Furthermore, silencing of the PHD2 gene with small interfering (si)RNAs abolished the reduction in the expression of hypoxia-inducible factor (HIF)-1α and VEGF by TMP. In addition, TMP protected CoCl2-induced HUVEC injury via an apoptosis pathway, as characterized by the increased ratio of cell viability and the reduced percentage of apoptotic and terminal deoxynucleotidyl transferase dUTP nick end labeling-positive HUVECs, activation of caspase-3, -8 and -9, B-cell lymphoma (Bcl)-2/Bcl-2-activated X protein expression, as well as the release of cytochrome c. The protective properties of TMP were partially attributed to the mRNA and protein expression levels of PHD, since silencing of the PHD2 gene with siRNAs abolished these effects. The present study demonstrated that the antiapoptotic effect of TMP in CoCl2-induced HUVECs was, at least in part, via the regulation of the PHD2/HIF-1α signaling pathway.

Curcumin pretreatment and post-treatment both improve the antioxidative ability of neurons with oxygen-glucose deprivation.

Recent studies have shown that induced expression of endogenous antioxidative enzymes thr-ough activation of the antioxidant response element/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway may be a neuroprotective strategy. In this study, rat cerebral cortical neurons cultured in vitro were pretreated with 10 µM curcumin or post-treated with 5 µM curcumin, respectively before or after being subjected to oxygen-glucose deprivation and reoxygenation for 24 hours. Both pretreatment and post-treatment resulted in a significant decrease of cell injury as indicated by propidium iodide/Hoechst 33258 staining, a prominent increase of Nrf2 protein expression as indicated by western blot analysis, and a remarkable increase of protein expression and enzyme activity in whole cell lysates of thioredoxin before ischemia, after ischemia, and after reoxygenation. In addition, post-treatment with curcumin inhibited early DNA/RNA oxidation as indicated by immunocytochemistry and increased nuclear Nrf2 protein by inducing nuclear accumulation of Nrf2. These findings suggest that curcumin activates the expression of thioredoxin, an antioxidant protein in the Nrf2 pathway, and protects neurons from death caused by oxygen-glucose deprivation in an in vitro model of ischemia/reperfusion. We speculate that pharmacologic stimulation of antioxidant gene expression may be a promising approach to neuroprotection after cerebral ischemia.