Glycyrrhizic acid treatment ameliorates anxiety-like behaviour via GLT1 and Per1/2-dependent pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: As a traditional Chinese medicinal herb, Glycyrrhiza. URALENSIS: Fisch. (licorice root, chinese name: Gancao) has a variety of medicinal values and is widely used clinically. Its main active ingredient, glycyrrhizic acid (GA), is believed to have a neuroprotective effect. However, the underlying biological mechanisms of GA on stress-induced anxiety disorders are still unclear. AIM OF THE STUDY: To investigate the anti-anxiety effect of GA and its underlying mechanism. METHODS: We selected the anxiety model induced by repeated chronic restraint stress (CRS) for 2 h on each of 7 consecutive days. GA (4, 20, 100 mg/kg) was injected intraperitoneally once daily for 1 week. The potential GA receptors were identified using whole-cell patches and computer-assisted docking of molecules. High-throughput RNA sequencing, adeno-associated virus-mediated gene regulation, Western blotting, and RT-qPCR were used to assess the underlying molecular pathways. RESULTS: GA alleviate depression-like and anxiety-like behaviors in CRS mice. GA decreased synaptic transmission by facilitating glutamate reuptaking in mPFC. Meanwhile, long-term GA treatment increased the expression of clock genes Per1 and Per2. Suppressing both Per1 and Per2 abolished the anxiolytic effects of GA treatment. CONCLUSION: Our study suggests that GA may be developed for the treatment of stress-induced anxiety disorders, and its mechanism is related to GLT1 and Per1/2-dependent pathways. This presents a novel approach to discovering potent therapeutic drugs.

Evaluating early lymphocyte-to-monocyte ratio as a predictive biomarker for delirium in older adult patients with sepsis: insights from a retrospective cohort analysis.

Background: This study aims to explore the value of the Lymphocyte-to-Monocyte Ratio (LMR) in predicting delirium among older adult patients with sepsis. Methods: Retrospective data were obtained from the MIMIC-IV database in accordance with the STROBE guidelines. Patients aged 65 and above, meeting the Sepsis 3.0 criteria, were selected for this study. Delirium was assessed using the Confusion Assessment Method for the ICU (CAM-ICU). Demographic information, comorbid conditions, severity of illness scores, vital sign measurements, and laboratory test results were meticulously extracted. The prognostic utility of the Lymphocyte-to-Monocyte Ratio (LMR) in predicting delirium was assessed through logistic regression models, which were carefully adjusted for potential confounding factors. Results: In the studied cohort of 32,971 sepsis patients, 2,327 were identified as meeting the inclusion criteria. The incidence of delirium within this subgroup was observed to be 55%. A univariate analysis revealed a statistically significant inverse correlation between the Lymphocyte-to-Monocyte Ratio (LMR) and the risk of delirium (p < 0.001). Subsequent multivariate analysis, which accounted for comorbidities and illness severity scores, substantiated the role of LMR as a significant predictive marker. An optimized model, achieving the lowest Akaike Information Criterion (AIC), incorporated 17 variables and continued to demonstrate LMR as a significant prognostic factor (p < 0.01). Analysis of the Receiver Operating Characteristic (ROC) curve indicated a significant enhancement in the Area Under the Curve (AUC) upon the inclusion of LMR (p = 0.035). Conclusion: The Lymphocyte-to-Monocyte Ratio (LMR) serves as a significant, independent prognostic indicator for the occurrence of delirium in older adult patients with sepsis. Integrating LMR into existing predictive models markedly improves the identification of patients at elevated risk, thereby informing and potentially guiding early intervention strategies.

Elucidation of the mechanism of action of ailanthone in the treatment of colorectal cancer: integration of network pharmacology, bioinformatics analysis and experimental validation.

Background: Ailanthone, a small compound derived from the bark of Ailanthus altissima (Mill.) Swingle, has several anti-tumour properties. However, the activity and mechanism of ailanthone in colorectal cancer (CRC) remain to be investigated. This study aims to comprehensively investigate the mechanism of ailanthone in the treatment of CRC by employing a combination of network pharmacology, bioinformatics analysis, and molecular biological technique. Methods: The druggability of ailanthone was examined, and its targets were identified using relevant databases. The RNA sequencing data of individuals with CRC obtained from the Cancer Genome Atlas (TCGA) database were analyzed. Utilizing the R programming language, an in-depth investigation of differentially expressed genes was carried out, and the potential target of ailanthone for anti-CRC was found. Through the integration of protein-protein interaction (PPI) network analysis, GO and KEGG enrichment studies to search for the key pathway of the action of Ailanthone. Then, by employing molecular docking verification, flow cytometry, Transwell assays, and Immunofluorescence to corroborate these discoveries. Results: Data regarding pharmacokinetic parameters and 137 target genes for ailanthone were obtained. Leveraging The Cancer Genome Atlas database, information regarding 2,551 differentially expressed genes was extracted. Subsequent analyses, encompassing protein-protein interaction network analysis, survival analysis, functional enrichment analysis, and molecular docking verification, revealed the PI3K/AKT signaling pathway as pivotal mediators of ailanthone against CRC. Additionally, the in vitro experiments indicated that ailanthone substantially affects the cell cycle, induces apoptosis in CRC cells (HCT116 and SW620 cells), and impedes the migration and invasion capabilities of these cells. Immunofluorescence staining showed that ailanthone significantly inhibited the phosphorylation of AKT protein and suppressed the activation of the PI3K/AKT signaling pathway, thereby inhibiting the proliferation and metastasis of CRC cells. Conclusion: Therefore, our findings indicate that Ailanthone exerts anti-CRC effects primarily by inhibiting the activation of the PI3K/AKT pathway. Additionally, we propose that Ailanthone holds potential as a therapeutic agent for the treatment of human CRC.

Investigating the mechanism of Sinisan formula in depression treatment: a comprehensive analysis using GEO datasets, network pharmacology, and molecular docking.

The herbal formula Sinisan (SNS) is a commonly used treatment for depression; however, its mechanism of action remains unclear. This article uses a combination of the GEO database, network pharmacology and molecular docking technologies to investigate the mechanism of action of SNS. The aim is to provide new insights and methods for future depression treatments. The study aims to extract effective compounds and targets for the treatment of depression from the T CMSP database. Relevant targets were searched using the GEO, Disgenet, Drugbank, PharmGKB and T T D databases, followed by screening of core targets. In addition, GO and KEGG pathway enrichment analyses were performed to explore potential pathways for the treatment of depression. Molecular docking was used to evaluate the potential targets and compounds and to identify the optimal core protein-compound complex. Molecular dynamics was used to further investigate the dynamic variability and stability of the complex. The study identified 118 active SNS components and 208 corresponding targets. Topological analysis of P P I networks identified 11 core targets. GO and KEGG pathway enrichment analyses revealed that the mechanism of action for depression involves genes associated with inflammation, apoptosis, oxidative stress, and the MAP K3 and P I3K-Akt signalling pathways. Molecular docking and dynamics simulations showed a strong binding affinity between these compounds and the screened targets, indicating promising biological activity. The present study investigated the active components, targets and pathways of SNS in the treatment of depression. Through a preliminary investigation, key signalling pathways and compounds were identified. These findings provide new directions and ideas for future research on the therapeutic mechanism of SNS and its clinical application in the treatment of depression.Communicated by Ramaswamy H. Sarma.

Network pharmacology and molecular docking to study the potential molecular mechanism of Qi Fu Yin for diabetic encephalopathy.

Diabetic encephalopathy is a chronic complication of diabetes that lacks an optimized treatment strategy. The present study sought to elucidate the potential molecular mechanism of Qi Fu Yin in improving diabetic encephalopathy through network pharmacology. The active components and target information of Qi Fu Yin were obtained from the TCMSP and Swiss target databases, while the target information of diabetic encephalopathy was sourced from Gene cards, OMIM, and Pharm Gkb databases. Enrichment analyses of KEGG and GO were conducted utilizing drug-disease common targets, while protein-protein interactions were predicted through the utilization of the STRING database platform. Subsequently, molecular docking was executed via Auto Dock Vina to authenticate the interaction between core components and core targets. The findings revealed that Qi Fu Yin exhibited 178 common targets with diabetic encephalopathy, and the enrichment analyses demonstrated that these targets were associated with lipid and atherosclerosis, AGE-RAGE signaling pathways, and other related pathways. The findings of the molecular docking indicated a favorable binding affinity between the active components of drug and the core targets, with EGF and quercetin exhibiting the most notable docking score. Additionally, the molecular dynamics simulation corroborated this high affinity. These results suggested that the active ingredients of Qi Fu Yin, including quercetin and kaempferol, may modulated the expression of genes such as IL10, TNF, EGF, and MMP2, thereby activating the AGE-RAGE signaling pathways and potentially serving as a therapeutic intervention for diabetic encephalopathy.Communicated by Ramaswamy H. Sarma.

The prolonged effect of film mulch and P application on lucerne forage yield in a semiarid environment.

Introduction: Limited water and soil phosphorus (P) availability often hampers lucerne productivity in semiarid regions. Plastic film mulch and P application typically enhance young lucerne (2-3 years) productivity by increasing soil water use and P availability. However, the prolonged impact of film mulch and P application on lucerne productivity as the stand ages remains unclear. Methods: This study conducted a 9-year field experiment on the semiarid Loess Plateau to investigate how film mulch and P application affect lucerne forage yield, soil water content, and soil fertility. The field experiment used a split-plot design with randomized blocks, in which the whole plots were with (M1) and without plastic film mulch (M0), and the split plots were four P rates (0 (P0), 9.7 (P1), 19.2 (P2), and 28.8 (P3) kg P ha-1). Results and discussion: The M1 treatment produced significantly higher lucerne forage yields than the M0 treatment during the first five years, but the yield-increasing effect of film mulch gradually diminished over time, with no effect in Years 6-8, and lower yields than the M0 treatment in Year 9. Phosphorus fertilization significantly increased forage yield after Year 3 in the M0 treatment, but only in Years 3-5 in the M1 treatment. In Years 2-5, film mulch significantly increased soil organic carbon, total nitrogen (N), inorganic N, and microbial biomass carbon in P0, P1, and P2 but not in P3. However, in Years 7-9, film mulch significantly decreased soil available potassium (K), organic carbon mineralization, lucerne density, and shoot K concentration, but did not reduce soil N and P availability at any level P of application. Moreover, plastic film mulch significantly increased the soil water content at 0-300 cm deep from Year 7 onwards. In conclusion, film mulch ceased to enhance lucerne production beyond year 6, which could not be attributed to soil water content, N or P availability but was partially associated with reduced soil K availability. Consequently, future research should focus on soil K availability, and K addition should be considered after five years in lucerne pastures mulched with plastic film in semiarid areas.

Virtual screening-molecular docking-activity evaluation of Ailanthus altissima (Mill.) swingle bark in the treatment of ulcerative colitis.

BACKGROUND: The dried bark of Ailanthus altissima (Mill.) Swingle is widely used in traditional Chinese medicine for the treatment of ulcerative colitis. The objective of this study was to explore the therapeutic basis of the dried bark of Ailanthus altissima (Mill.) Swingle for the treatment of ulcerative colitis based on Virtual Screening-Molecular Docking-Activity Evaluation technology. METHODS: By searching the Traditional Chinese Medicine Systems Pharmacology TCMSP Database and Analysis Platform, 89 compounds were obtained from the chemical components of the dried bark of Ailanthus altissima (Mill.) Swingle. Then, after preliminarily screening the compounds based on Lipinski's rule of five and other relevant conditions, the AutoDock Vina molecular docking software was used to evaluate the affinity of the compounds to ulcerative colitis-related target proteins and their binding modes through use of the scoring function to identify the best candidate compounds. Further verification of the compound's properties was achieved through in vitro experiments. RESULTS: Twenty-two compounds obtained from the secondary screening were molecularly docked with ulcerative colitis-related target proteins (IL-1R, TLR, EGFR, TGFR, and Wnt) using AutoDock Vina. The free energies of the highest scoring compounds binding to the active cavity of human IL-1R, TLR, EGFR, TGFR, and Wnt proteins were - 8.7, - 8.0, - 9.2, - 7.7, and - 8.5 kcal/mol, respectively. The potential compounds, dehydrocrebanine, ailanthone, and kaempferol, were obtained through scoring function and docking mode analysis. Furthermore, the potential compound ailanthone (1, 3, and 10 µM) was found to have no significant effect on cell proliferation, though at 10 µM it reduced the level of pro-inflammatory factors caused by lipopolysaccharide. CONCLUSION: Among the active components of the dried bark of Ailanthus altissima (Mill.) Swingle, ailanthone plays a major role in its anti-inflammatory properties. The present study shows that ailanthone has advantages in cell proliferation and in inhibiting of inflammation, but further animal research is needed to confirm its pharmaceutical potential.

The ability of long non-coding RNA RP11-284N8.3 to predict the risk, the severity and 28-day mortality of adults with sepsis.

In a prior study, we identified a novel sepsis specific long noncoding RNAs (lncRNA) RP11-284N8.3, which may primarily participate in T cell activation and immune response during sepsis. However, the clinical significance of lncRNA RP11-284N8.3 in sepsis remains entirely unknown. This single-center prospective cohort study enrolled 147 adults with sepsis and 74 healthy controls (HCs) with matched age and sex between January 2021 and November 2022 at our hospital. Blood samples and clinical data were collected from HCs at enrollment and from adults with sepsis within 24 hours after admission. lncRNA RP11-284N8.3 expression was detected by RT-qPCR. The relative expression of lncRNA RP11-284N8.3 was significantly decreased in adults with sepsis compared to HCs (P < .0001), in adults with septic shock compared to adults without shock (P = .0012), and in 28-day deaths compared to 28-day survivors (P = .0006). receiver operating characteristic curves of lncRNA RP11-284N8.3 in predicting sepsis severity and 28-day mortality showed an area under the curve of 0.6570 (95% confidence interval [CI]: 0.5701-0.7440) and an area under the curve of 0.6765 (95% CI: 0.5809-0.7721), respectively. Multivariate logistic regression analysis revealed that lncRNA RP11-284N8.3 was an independent risk factor for 28-day mortality in adults with sepsis (odds ratio: 0.1057, 95% CI: 0.0115-0.7746, P = .0328). Low expression of lncRNA RP11-284N8.3 is correlated with increased risk, severity and 28-day mortality in adults with sepsis, and it may function as a potential biomarker to facilitate the diagnosis and management of sepsis.

Inflammatory Factors Predicted the Resilient Phenotype in Social Defeat-induced Depression of Male Mouse.

An absence of reward in chronic stress may impair the reward circuit in the brain, resulting in major depressive disorder (MDD). In a part of chronically stressed individuals, MDD is not present, i.e., there is resilience, implying endogenous anti-depressive mechanisms in the brain. We studied social defeat model mice and analyzed the mRNA maps of the hippocampus from a control group and social defeat (SD)-susceptible and SD-resilient mice using high-throughput sequencing techniques. It was found that the immune response was associated with depression. Existing studies have proven that microglia play an important role in the brain immune response, and their activation level increases after chronic social defeat stress (CSDS). In our study, minocycline inhibited the activation of microglia, thereby improving the depressive state of CSDS mice. In addition, minocycline combined with fluoxetine enhanced the efficacy of fluoxetine. Thus, our results propose the most probable mechanism underlying different responses to CSDS and indicate the potential of a combination of anti-inflammatory drugs and antidepressants in treating refractory depression.

Investigation into the in vivo mechanism of diosmetin in patients with breast cancer and COVID-19 using bioinformatics.

Patients with breast cancer are prone to SARS-CoV-2 infection [the causative virus of coronavirus disease (COVID-19)] due to their lack of immunity. In the current study, we examined the mechanism of action of Diosmetin, a flavonoid with anti-inflammatory properties, in patients with BRCA infected with SARS-CoV-2.We used bioinformatics technology to analyze the binding ability, biological function, and other biological characteristics of Diosmetin in vivo and examine the core target and potential mechanism of action of Diosmetin in patients with patients with breast cancer infected with SARS-CoV-2. A prognostic model of SARS-COV-2-infected breast cancer patients was constructed, and the core genes were screened out, revealing the correlation between these core genes and clinicopathological characteristics, survival rate, and high-risk and low-risk populations. The docking results revealed that Diosmetin binds well to the core genes of patients with breast cancer with COVID-19. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses suggested that Diosmetin inhibited inflammation, enhanced immune function, and regulated the cellular microenvironment in patients with BRCA/COVID-19. For the first time, we reveal the molecular functions and potential targets of Diosmetin in patients with breast cancer infected with SARS-CoV-2, improving the reliability of the new drug and laying the foundation for further research and development.

The effects of plastic film mulching and straw mulching on licorice root yield and soil organic carbon content in a dryland farming.

The increasing worldwide demand for traditional herbs has been met by growing cultivated herbs. It is undoubtedly very important to seek a reasonable cultivation mode for the yield, quality and long-term production stability of traditional herbs. In this study, licorice (Glycyrrhiza uralensis Fisch.) was investigated using a field experiment and a process-based model (Denitrification-Decomposition (DNDC) model) to study the effects of mulching methods on root yield and soil organic carbon (SOC) long-term changes. The field experiment contained four treatments: plat planting without mulching (CK), ridge-furrow maize straw mulching (SM), ridge-furrow plastic film mulching (RP), and plat planting with plastic film mulching (FP). Licorice root yield was significantly higher in the SM, RP, and FP than in the CK. SM, RP and FP treatments increased the accumulation of liquiritin and glycyrrhizin in licorice roots. The SM significantly increased SOC content, SOC stocks, SOC sequestration rate, dissolved organic carbon (DOC) content and microbial biomass carbon (MBC) compared to CK, but there was no significant difference in SOC and DOC among CK, RP and FP. The DNDC model was calibrated based on the field test data and showed that under the four CMIP6 SSPs scenarios, the predicted root yield of each treatment was increasing obviously. The production and stability of RP and FP were greater than CK and SM. The SOC under SM showed an increasing trend, whereas it continuously decreased under CK, RP, and FP in the future. The SOC of simulated RS treatment of straw incorporation plus a plastic film mulch was always at the highest value in all the treatments, and its root yield was slightly lower than that of RP and FP, the latter both were very close. Therefore, it is suggested that RS should be adopted to achieve sustained high yield while maintaining a high SOC level.

Reactive Oxygen Species-Responsive Polyether Micelle Nanomaterials for Targeted Treatment of Ulcerative Colitis.

As one of the most challenging inflammatory diseases, the incidence of ulcerative colitis (UC) is increasing year by year, but the existing therapeutic drugs are not effective and lack of targeting. Nanomaterials are expected to become promising delivery system due to their good targeting effects. Here, we designed a nanomaterial sensitive to reactive oxygen species, which can be used to treat IBD, especially UC. It is a self-assembled polyether micelle that can be oxidized at the inflammation site where the concentration of reactive oxygen increases, and effectively release the encapsulated budesonide (Bud). Experiments have proved that for DSS-induced colitis, the synthetic drug-loaded nanoparticles have excellent therapeutic effects, can effectively repair intestinal barrier, and significantly improve the damaged colon tissue. At the same time, it has a beneficial regulatory effect on inflammatory factors. Molecular mechanism studies have shown that it achieves its therapeutic effects by activating the peroxisome proliferators-activated receptors-γ (PPAR-γ) pathway and inhibiting the nuclear factor (NF)-κB pathway. This study proves that oral nano-micelles have an important impact on improving the efficacy of UC treatment drugs and have far-reaching significance for the targeted treatment of gastrointestinal diseases.

Xanthohumol protect cognitive performance in diabetic model rats by inhibiting protein kinase B/nuclear factor kappa-B pathway.

Xanthohumol (XN, 2', 4', 4-trihydroxy-6'-methoxy-3'-prenylchalcone), a polyphenol chalcone from hops (Humulus lupulus), has received increasing attention due to its multiple pharmacologic activities. As an active component in beers, its presence has been suggested to be linked to the epidemiologic observation of the beneficial effect of regular beer drinking. But regarding cardiovascular and immunologic effects of polyphenols and ethanol, benefits of beer drinking in patients with diabetes were still in doubt. Diabetes was induced in male Sprague-Dawley rats by administering a high-fat diet and an intraperitoneal 30 mg/kg streptozotocin injection. The animals were treated orally with saline or XN at 50 mg/kg/d for 4 weeks. At the end of the treatment, hippocampus from different groups were collected for biochemical examination. In this study, we found XN inhibit phosphorylation of protein kinase B and nuclear factor kappa-B which was overactivated in diabetic rats, followed by decreased blood glucose and increased body weight. Additionally, XN treatment significantly increased freezing time in a fear memory test. In further research, we found XN increased synaptic plasticity and dendritic spine density, while decreased reactive oxygen species in hippocampus slices from diabetic rats. All these results indicate that XN might be a promising drug to treat diabetic encephalopathy.

[Value of interleukin-6 and CD4+ T-lymphocytopenia in assessing the severity and prognosis of coronavirus disease 2019].

OBJECTIVE: To evaluate the role of interleukin-6 (IL-6) and CD4+ T-lymphocytopenia in assessing the severity and prognosis of coronavirus disease 2019 (COVID-19). METHODS: A prospective observational study was conducted. Forty-five patients with COVID-19 admitted to Henan Provincial People's Hospital from January 13 to March 13, 2020 were enrolled and divided into normal group (13 cases), severe group (20 cases), critically severe group (12 cases) according to the severity of the disease. A total of 15 healthy subjects receiving physical examinations during the same period were collected as the healthy control group. Clinical data were collected to compare the clinical characteristics, general test results, IL-6 and CD4+ T-lymphocytopenia levels of patients in different disease severity groups and healthy control group. The receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of each indicator for the severity of COVID-19. Multivariate Cox regression analysis was used to analyze the risk factors affecting the prognosis of COVID-19 patients, and Kaplan-Meier survival curve analysis was performed. RESULTS: The age of the critically severe group was significantly higher than that of the severe and normal groups (years old: 66.91±17.01 vs. 59.35±18.07, 40.23±12.61, both P < 0.05), and the negative conversion time of the 2019 novel coronavirus (2019-nCoV) was significantly longer than that of the severe and normal groups (days: 19.00±10.66 vs. 18.00±7.18, 9.31±3.49, both P < 0.05). With the increase of the severity of disease, white blood cell count (WBC), C-reactive protein (CRP), calcitonin (PCT), total bilirubin (TBil), troponin I (TnI), IL-6, D-dimer and other indicators were significantly increased, while lymphocyte count (LYM), platelet count (PLT), CD4+, CD8+, oxygenation index (PaO2/FiO2) were significantly decreased (all P < 0.01). ROC curve showed that PaO2/FiO2, IL-6 and CD4+ had certain predictive value for disease severity of COVID-19, the area under the ROC curve (AUC) of them were 0.903, 0.871, 0.689, and the 95% confidence interval (95%CI) were 0.806-0.949, 0.769-0.974, 0.542-0.853; the best cut-off values were 196.00 mmHg (1 mmHg = 0.133 kPa), 6.02 ng/L, 355 cells/µL, respectively; the sensitivity were 73.3%, 99.3%, 73.3%, and the specificity were 96.6%, 62.1%, 65.5%, respectively. Multivariate Cox regression analysis showed that age, PaO2/FiO2, high IL-6 and low CD4+ (IL-6 ≥ 6.02 ng/L and CD4+ < 355 cells/µL) were independent risk factors affecting the prognosis of COVID-19 [hazard ratio (HR) was 1.077, 0.053 and 3.490, respectively, all P < 0.05]. Kaplan-Meier survival analysis showed that when both high IL-6 and low CD4+ (IL-6 ≥ 6.02 ng/L and CD4+ < 355 cells/µL) were present, the mean time of adverse prognosis was (20.53±5.71) days; when increased IL-6 and decreased CD4+ were inconsistent, the mean time of adverse prognosis was (53.21±3.16) days. CONCLUSIONS: The levels of IL-6 and CD4+ T-lymphocytopenia are closely related to the severity of COVID-19 disease. When IL-6 ≥ 6.02 ng/L and CD4+ < 355 cells/µL occur simultaneously, the prognosis is poor.

Sustained Release of Melatonin from GelMA Liposomes Reduced Osteoblast Apoptosis and Improved Implant Osseointegration in Osteoporosis.

A reduction in bone mass around an implant is the main cause of implant loosening, especially in postmenopausal osteoporosis patients. In osteoporosis, excessive oxidative stress, resulting in osteoblast apoptosis, largely contributes to abnormal bone remodeling. Melatonin (MT) synthesized by the pineal gland promotes osteoblast differentiation and bone formation and has been effectively used to combat oxidative stress. Therefore, we hypothesized that MT attenuates osteoblast apoptosis induced by oxidative stress, promotes osteogenesis in osteoporosis, and improves bone mass around prostheses. Moreover, considering the distribution and metabolism of MT, its systemic administration would require a large amount of MT, increasing the probability of drug side effects, so the local administration of MT is more effective than its systemic administration. In this study, we constructed a composite adhesive hydrogel system (GelMA-DOPA@MT) to bring about sustained MT release in a local area. Additionally, MT-reduced apoptosis caused by hydrogen peroxide- (H2O2-) induced oxidative stress and restored the osteogenic potential of MC3T3-E1 cells. Furthermore, apoptosis in osteoblasts around the implant was significantly attenuated, and increased bone mass around the implant was observed in ovariectomized (OVX) rats treated with this composite system. In conclusion, our results show that GelMA-DOPA@MT can inhibit osteoblast apoptosis caused by oxidative stress, thereby promoting osteogenesis and improving bone quality around a prosthesis. Therefore, this system of local, sustained MT release is a suitable candidate to address implant loosening in patients with osteoporosis.

[Ruyi Jinhuang Plaster suppresses BPH by reducing the expressions of P38, JNK2, NF-кBP65 and STAT3 in the prostate tissue of rats].

OBJECTIVE: To investigate the effect of Ruyi Jinhuang Plaster (RJP) on testosterone propionate-induced BPH in the rat model and its action mechanisms. METHODS: Forty-eight SD male rats were randomly divided into six groups of equal number: normal control, BPH model control, finasteride, and high-, medium- and low-dose RJP. The BPH model was made in the latter five groups by hypodermic injection of testosterone propionate. From the first day of modeling, the rats of the normal control and BPH model control groups were treated with blank plasters and those of the high-, medium- and low-dose RJP groups with RJPs at 42.0, 21.0 and 10.5 cm2/kg applied to the dehaired area of the back, and those of the finasteride group by gavage of finasteride at 4.5 mg/kg, all once a day for 30 successive days. Then the prostates of the animals were harvested for observation of histopathological changes by HE staining, measurement of the areas of interstitial and epithelial cells and prostatic glandular cavity, and determination of the expressions of P38, JNK2, NF-кBP65 and STAT3 proteins in the prostate tissue by Western blot. RESULTS: Compared with the BPH model controls, the high-dose RJP group showed significantly decreased proliferation and area proportion of prostatic epithelial cells (P < 0.05), increased area proportion of the prostatic glandular cavity (P < 0.05), and reduced expressions of P38, p-P38, NF-кBP65, P-NF-кBP65, STAT3, P-STAT3 and JNK2 in the prostate tissue (P < 0.05); the medium-dose RJP group exhibited markedly down-regulated expressions of JNK2 and NF-кBP65 (P < 0.05) but an up-regulated level of p-JNK (P < 0.05); while the low-dose RJP group displayed a remarkably reduced expression of JNK2 (P < 0.05) but an elevated level of p-JNK (P < 0.05). CONCLUSIONS: RJP suppresses BPH in the model rat by down-regulating the expressions of P38, p-P38, NF-кBP65, P-NF-кBP65, STAT3, P-STAT3 and JNK2 or up-regulating that of p-JNK in the prostate tissue.

Epigallocatechin-3-gallate ameliorates insulin resistance in hepatocytes.

Hyperglycemia is a typical pathogenic factor in a series of complications among patients with type II diabetes. Epigallocatechin-3-gallate (EGCG) is the major polyphenol extracted from green tea and is reported to be an antioxidant. The aim of the present study was to examine the effect of EGCG on insulin resistance in human HepG2 cells pretreated with high concentrations of glucose. The protein kinase B (AKT)/glycogen synthase kinase (GSK) pathways were analyzed using western blot analysis in HepG2 cells and primary mouse hepatocytes treated with high glucose and/or EGCG. Cellular glycogen content was determined using a glycogen assay kit. Reactive oxygen species (ROS) production was determined using dihydroethidium staining and flow cytometry. c­JUN N­terminal kinase (JNK)/insulin receptor substrate 1 (IRS1)/AKT/GSK signaling was explored using western blot analysis in HepG2 cells treated with high glucose and/or EGCG or N-acetyl-cysteine. High glucose significantly decreased the levels of phosphorylated AKT and GSK in HepG2 cells and mouse primary hepatocytes. Pretreatment with EGCG significantly restored the activation of AKT and GSK in HepG2 cells and primary hepatocytes exposed to high glucose. In HepG2 cells and primary hepatocytes, glycogen synthesis was improved by EGCG treatment in a dose­dependent manner. High glucose significantly stimulated the production of ROS while EGCG protected high glucose­induced ROS production. ROS is known to serve a major role in high glucose induced­insulin resistance by increasing JNK and IRS1 serine phosphorylation. In the present study, EGCG was observed to enhance the insulin­signaling pathway. EGCG ameliorated high glucose­induced insulin resistance in the hepatocytes by potentially decreasing ROS­induced JNK/IRS1/AKT/GSK signaling.

[Value of serum miR-155-5p and miR-133a-3p expression for the diagnosis and prognosis evaluation of sepsis].

OBJECTIVE: To explore the value of serum microRNA-155-5p and -133a-3p (miR-155-5p and miR-133a-3p) expression for the diagnosis and prognosis evaluation of sepsis. METHODS: A prospective observational study was conducted. 105 sepsis patients admitted to emergency intensive care unit (EICU) of the First Affiliated Hospital of Zhengzhou University from January 2015 to January 2016 were enrolled. They were divided into three groups according to the severity: 35 patients with sepsis, 35 with severe sepsis, and 35 with septic shock. At the same time, 35 healthy persons were selected as the control group. According to the prognosis, the patients were divided into improved group (n = 70) and in-hospital death group (n = 35). The clinical data of all the subjects were collected. The mRNA expressions of miR-155-5p and miR-133a-3p were determined by reverse transcription-polymerase chain reaction (RT-PCR). The receiver-operating characteristic curve (ROC) was plotted to evaluate their clinical value for the diagnosis and prognosis of sepsis. The binary logistic regression was used to analyze the risk factors affecting the prognosis of sepsis patients. RESULTS: (1) The mRNA expressions of serum miR-155-5p and miR-133a-3p were gradually increased with the aggravation of sepsis. The mRNA expression of miR-155-5p (2-ΔCt) in sepsis, severe sepsis, sepsis shock groups was 1.89±0.48, 2.21±0.41, 2.79±0.73 (F = 23.737, P = 0.000), and the mRNA expression of miR-133a-3p (2(-ΔCt)) was 1.38±0.31, 1.74±0.65, 2.08±0.47, respectively (F = 27.710, P = 0.000). It was shown by ROC curve analysis that the area under the ROC curve (AUC) of serum miR-155-5p and miR-133a-3p for the diagnosis of sepsis was 0.855 [95% confidence interval (95%CI) = 0.761-0.949] and 0.769 (95%CI = 0.666-0.872) respectively. The cut-off value of miR-155-5p for the diagnosis of sepsis was 1.64, the sensitivity was 85.3%, and specificity was 80.6%. While the cut-off value of miR-133a-3p was 0.82, the sensitivity and specificity were 97.9% and 54.8% respectively. (2) Compared with improved group, the patients of in-hospital death group were more serious, and procalcitonin (PCT), C-reactive protein (CRP), D-dimer, lactic acid (Lac), sequential organ failure assessment (SOFA) score, acute physiology and chronic health evaluation II (APACHE II) score, and the mRNA expressions of miR-155-5p and miR-133a-3p were significantly increased (all P < 0.05). While there was no statistically significant difference in gender, age, white blood cells (WBC), serum creatinine (SCr) between the two groups (all P > 0.05). It was shown by binary logistic regression analysis that Lac [odds ratio (OR) = 0.514, 95%CI = 0.260-0.893, P = 0.024], sepsis severity (OR = 0.039, 95%CI = 0.023-2.955, P = 0.016), SOFA score (OR = 0.668, 95%CI = 0.474-0.825, P = 0.001), serum miR-155-5p expression (OR = 0.117, 95%CI = 0.020-0.530, P = 0.007) were the risk factors affecting the prognosis of patients with sepsis. CONCLUSIONS: The expression of serum miR-155-5p and miR-133a-3p may be used as specific indicators for the diagnosis of sepsis. And the expression of miR-155-5p can be used as independent impact factor for the estimation of sepsis prognosis.

Huangqi-Honghua combination and its main components ameliorate cerebral infarction with Qi deficiency and blood stasis syndrome by antioxidant action in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Combination of Radix Astragali (Huangqi) and Carthamus tinctorius L. (Honghua) has been extensively used as traditional herb medicine in China for the treatment of stroke and myocardial ischemia diseases with Qi deficiency and blood stasis (QDBS) syndrome. AIM: To investigate the effect of Huangqi-Honghua combination (HH) and its main components astragaloside IV (AS-IV) and Hydroxysafflor yellow A (HSYA) on cerebral ischemia-reperfusion (IR) with QDBS in rat model. MATERIALS AND METHODS: Male rats were randomly divided into the following six groups: sham group, QDBS+I/R model group and treatment group including AS-IV, HSYA, AS-IV+HSYA and HH. The whole blood viscosity (WBV), plasma viscosity (PV), neurological examination, infarct volume, histopathology changes and some oxidative stress markers were assessed after 24h of reperfusion. RESULTS: HH and its main components AS-IV+HSYA could significantly decrease WBV, PV, and also significantly ameliorate neurological examination and infarct volume after 24h of reperfusion. They also significantly increased expression of Nuclear factor erythroid 2-related factor 2 (Nrf2), activities of antioxidants, such as superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px), led to decrease levels of malondialdehyde (MDA) and reactive oxygen species (ROS). CONCLUSION: AS-IV and HSYA are responsible for the main curative effects of HH. The study may provide scientific information to further understanding the mechanism(s) of HH and its main components in removing blood stasis and ameliorating cerebral infarction. Additionally, AS-IV and HSYA appear to have synergistic effects on neuroprotection.